The lichen genus Buellia de Not. in the greater Sonoran Desert Region: saxicolous species with one-septate ascospores

Frank Bungartz

Thirty-one saxicolous species of Buellia s.l. with one-septate ascospores were examined from the Greater Sonoran Desert Region. They are distinguished by thallus morphology, exciple anatomy, spore ultrastructure, secondary chemistry, ecology and distribution. As a result, seven new species are described. Other species are reported from the region for the first time and the synonymy of several taxa is resolved. All species are traditionally included within the Physciaceae. Recent molecular evidence suggests that calicioid lichens are closely related to Buellia s.l. A comparison confirms that spore wall ultrastructure is identical in both groups. Iodine staining of the ascus in the light microscope corresponds well with the ascus ultrastructure. This Bacidia-type ascus is variable but common to all species. The prototunicate asci of calicioid Physciaceae may have evolved by reduction from this type. Spore ontogeny of Buellia s.l. is a highly dynamic process. Four stages can be disting...

THE LICHEN GENUS BUELLIA DE NOT. IN THE GREATER SONORAN DESERT REGION: SAXICOLOUS SPECIES WITH ONE-SEPTATE ASCOSPORES by Frank Bungartz A Dissertation Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy ARIZONA STATE UNIVERSITY May 2004 THE LICHEN GENUS BUELLIA DE NOT. IN THE GREATER SONORAN DESERT REGION: SAXICOLOUS SPECIES WITH ONE-SEPTATE ASCOSPORES by Frank Bungartz has been approved May 2004 APPROVED: , Chair ________________________________________________________________________ ________________________________________________________________________ ________________________________________________________________________ Supervisory Committee ACCEPTED: ___________________________________ Director of the School ___________________________________ Dean, Graduate College dedicated to my wife Frauke for her love and support ABSTRACT Thirty-one saxicolous species of Buellia s.l. with one-septate ascospores were examined from the Greater Sonoran Desert Region. They are distinguished by thallus morphology, exciple anatomy, spore ultrastructure, secondary chemistry, ecology and distribution. As a result, seven new species are described. Other species are reported from the region for the first time and the synonymy of several taxa is resolved. All species are traditionally included within the Physciaceae. Recent molecular evidence suggests that calicioid lichens are closely related to Buellia s.l. A comparison confirms that spore wall ultrastructure is identical in both groups. Iodine staining of the ascus in the light microscope corresponds well with the ascus ultrastructure. This Bacidia-type ascus is variable but common to all species. The prototunicate asci of calicioid Physciaceae may have evolved by reduction from this type. Spore ontogeny of Buellia s.l. is a highly dynamic process. Four stages can be distinguished: (1) immature, (2) premature, (3) mature and (4) overmature. During these stages wall differentiation, septum formation and pigmentation develop independently. Wall thickenings result from endospore differentiation. They are most obvious in premature and mature spores, but usually become reduced with age. Exciple anatomy of Buellia s.l. is diagnostic and corresponds to some degree with pigmentation. Only B. mamillana has a thalline exciple, which soon becomes excluded. Other species lack a distinct thalline margin as observed in Rinodina s.l. Several inconspicuous species show some degree of substrate penetration but only Buellia sequax and B. vilis are chasmolithic. Pycnidia of all species are similar. A general transfer of taxa with filiform iii conidia into Amandinea is therefore not supported. Amandinea needs to be revised to include only species closely related to the type, Amandinea coniops. A phylogenetic analysis of eighty species demonstrates that most generic segregates of Buellia s.l. are presently not well supported. Groups like Diplotomma s.str. or Hafellia may be recognized but even these are not strongly supported. The low support is inherent to the small amount of characters available. As a consequence, genera should only be accepted if classical concepts are re-evaluated with molecular tools. iv ACKNOWLEDGEMENTS First and foremost I am grateful to my committee members: Dr. Thomas H. Nash III has been most influential as my Doktorvater. He has guided me and provided much helpful support whenever I needed it. I owe my microscopy skills to Dr. Robert W. Roberson. Dr. Donald J. Pinkava helped me deciphering the botanical code. Dr. Bruce D. Ryan has also been invaluable in this regard. His knowledge of lichens was inexhaustible. When he died, I lost a dear friend. I shared many productive discussions on the taxonomy of Buellia and Rinodina with Dr. John W. Sheard from the University of Saskatchewan. Finally, Dr. Martin F. Wojciechowski assisted with the phylogenetic analysis and was ready to sit in for Dr. Pinkava during my defense on very short notice. I will never forget the British Lichen Society workshops taught by Peter W. James. Dr. Christoph Scheidegger introduced me to his research on saxicolous species of Buellia. I also met with Dr. Rosemarie Honegger learning important details about electron microscopy of lichens. Dr. Helmut Mayrhofer and Ulrike Grube (University of Graz), were helpful correspondents, who shared their enormous knowledge about the Physciaceae. Dr. John A. Elix analyzed specimens with High Performance Liquid Chromatography. During a visit to Bayreuth, I began investigating the molecular taxonomy of the Buellia dispersa-group with Dr. Gerhard Rambold and Dr. Derek Peršoh. Dr. Alan M. Fryday supported my research during a visit to MSC. Many other lichenologists were also helpful: Julia Blaha, Dr. Christian Printzen, Dr. Matthias Schultz, Dr. Ulrik Søchting, Dr. Anders Nordin, Dr. Eva Barreno, Charis C. Bratt, Dr. Robert S. Egan, Kerry Knudsen, Dr. Shirley C. Tucker, Dr. H. Thorsten Lumbsch, Dr. Guido B. Feige, James C. Lendemer and Dr. Birgit Litterski. v This research would have been a lot more cumbersome without the excellent database at the ASU Lichen Herbarium, designed by Dr. Corinna Gries. Scott T. Bates has carefully reviewed most of my manuscripts. Other graduate students have also been very helpful: Robin T. Schoeninger, Raul Puente-Martinez, Ken G. Sweat, Karen L. Dillman and William A. Iselin. William P. Sharp, from the Electron Microscopy Facilities, was always available for microscopy questions. My research on endolithic lichens would have been impossible without Dr. Laurence A. Garvie from the Geology Department. Dr. Dennis McDaniel helped with confocal microscopy. The excellent staff at the School of Life Sciences maneuvered me through ASU as a foreign student. I have dedicated this dissertation to my wife, Frauke Ziemmeck. My parents, my sister and my friends all understand my passion for lichenology. I am unable to list many more individuals here. I nevertheless thank everyone, even without naming them. My research received financial support from: The Sonoran Lichen Flora Project (National Science Foundation awards DEB-0103738, DEB-9701111), a Research Grant in Plant Systematics from the International Association of Plant Taxonomists, a Research Grant of Sigma Xi chapter at ASU, several travel grants and other department funds. The following herbaria contributed specimens for my research: ASU, B, BM, C, CANL, CAS, COLO, FH, G, GZU, H, hb. Kalb, hb. Knudsen, hb. Lendemer, hb. Nordin, hb. Printzen, hb. Scheidegger, hb. van den Boom, L, M, MEXU, MICH, MIN, MSC, MU, OMA, PH, S, SASK, SBBG, STU, TUR, UC, UPS, US, W. vi TABLE OF CONTENTS Page LIST OF TABLES ......................................................................................................... xiv LIST OF FIGURES ....................................................................................................... xv INTRODUCTION ............................................................................................................ 1 MATERIAL AND METHODS ........................................................................................ 4 Specimens examined .................................................................................................... 4 Methods ........................................................................................................................ 4 LIGHT MICROSCOPY ............................................................................................. 4 SCANNING ELECTRON MICROSCOPY ..................................................................... 5 TRANSMISSION ELECTRON MICROSCOPY .............................................................. 5 SPOT TESTS ........................................................................................................... 5 THIN-LAYER CHROMATOGRAPHY ......................................................................... 5 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY ................................................ 6 CLADISTICS ........................................................................................................... 6 Parsimony Analysis ........................................................................................ 6 Character Coding .......................................................................................... 7 TAXONOMIC CONCEPTS ........................................................................................... 10 Taxonomic Concept of Buellia within the Physciaceae ............................................. 10 Taxonomic Concept of the Genus Buellia De Not. .................................................... 12 MORPHOLOGY AND ANATOMY ............................................................................. 16 Thallus Morphology ................................................................................................... 16 vii Page Thallus Anatomy ........................................................................................................ 19 Apothecia ................................................................................................................... 20 ANATOMY OF THE EXCIPLE AND HYPOTHECIUM ................................................. 20 The aethalea-type exciple ............................................................................. 25 The dispersa-type exciple ............................................................................. 30 The leptocline-type exciple .......................................................................... 30 The mamillana-type exciple ......................................................................... 31 The trachyspora-type exciple ........................................................................ 31 The vilis-type exciple ................................................................................... 32 Asci and Ascospores .................................................................................................. 32 ASCUS TYPE ........................................................................................................ 32 ASCOSPORES ....................................................................................................... 38 Spore Ultrastructure .................................................................................... 38 Spore Ontogeny and Spore Types ................................................................ 46 Towards a more dynamic concept of spore ontogeny in the Physciaceae ... 48 Conclusions .................................................................................................. 50 Pycnidia ...................................................................................................................... 50 CHEMISTRY ................................................................................................................. 56 Orcinol depsides ......................................................................................................... 58 Orcinol depsidones ..................................................................................................... 59 β-Orcinol depsides ..................................................................................................... 59 β-Orcinol depsidones ................................................................................................. 60 viii Page Xanthones ................................................................................................................... 60 Usnic acid related substances ..................................................................................... 60 Anthraquinones .......................................................................................................... 61 Other substances ......................................................................................................... 62 DISTRIBUTION AND ECOLOGY ............................................................................... 64 Distribution ................................................................................................................ 64 Ecology ...................................................................................................................... 79 PHYLOGENY ................................................................................................................ 82 Results of the Analysis ............................................................................................... 82 Discussion: Segregates from the Genus Buellia and Taxonomic Implications ......... 85 Amandinea M. Choisy ex Scheid. & M. Mayrhofer .................................... 87 Aplotomma A. Massal. ex Beltr. .................................................................. 90 Baculifera Marbach & Kalb ......................................................................... 91 Dimelaena Norman ...................................................................................... 92 Diploicia A. Massal. .................................................................................... 93 Diplotomma Flot. ......................................................................................... 93 Gassicurtia Fée ............................................................................................ 94 Hafellia Kalb, H. Mayrhofer & Scheid. ....................................................... 95 Karschia Körb. ............................................................................................. 96 Mattickiolichen Tomas. & Cif. .................................................................... 97 Melanaspicilia Vain. .................................................................................... 97 Monerolechia Trevis. ................................................................................... 98 ix Page Rinodina (Ach.) Gray ................................................................................. 101 Tetramelas Norman ................................................................................... 102 Buellia species with xanthones .................................................................. 102 Conclusions .............................................................................................................. 103 PUBLICATIONS .......................................................................................................... 105 PUBLICATION 1 (Bibliotheca Lichenologica 82, 2002): Buellia dispersa A. Massal., a variable lichen species from semi-arid to arid environments of North America and Europe ................................................................................................................. 105 MATERIALS AND METHODS .............................................................................. 108 RESULTS ........................................................................................................... 111 DISCUSSION ...................................................................................................... 124 ACKNOWLEDGEMENTS ...................................................................................... 133 LITERATURE CITED ........................................................................................... 134 PUBLICATION 2 (The Bryologist 107, 2004): Buellia turgescens is synonymous with Buellia badia and must not be included in Amandinea ....................................... 137 METHODS ......................................................................................................... 138 RESULTS AND DISCUSSION ................................................................................ 139 ACKNOWLEDGMENTS ........................................................................................ 153 LITERATURE CITED ........................................................................................... 153 PUBLICATION 3 (The Bryologist 107, 2004): Buellia saurina belongs to the genus Rhizocarpon (Rhizocarpaceae, Lichenized Ascomycetes) .................................. 156 ACKNOWLEDGMENTS ........................................................................................ 164 x Page LITERATURE CITED ........................................................................................... 164 PUBLICATION 4 (Bibliotheca Lichenologica 88, 2004): Buellia subalbula (Nyl.) Müll. Arg. and B. amabilis de Lesd., two species from North America with one-septate ascospores: A comparison with Buellia [“Diplotomma”] venusta (Körb.) Lettau. ... .............................................................................................................................. 166 METHODS ......................................................................................................... 169 RESULTS ........................................................................................................... 170 DISCUSSION ...................................................................................................... 189 CONCLUSIONS ................................................................................................... 195 ACKNOWLEDGEMENTS ...................................................................................... 195 LITERATURE CITED ........................................................................................... 196 PUBLICATION 5 (Canadian Journal of Botany, in press): Morphology and anatomy of chasmolithic versus epilithic growth: a taxonomic revision of inconspicuous saxicolous Buellia species from the Sonoran Desert Region generally ascribed to the “Buellia punctata”-group ............................................................................... 201 METHODS ......................................................................................................... 203 RESULTS ........................................................................................................... 204 Key to the species: ...................................................................................... 204 DISCUSSION ...................................................................................................... 239 ACKNOWLEDGEMENTS ...................................................................................... 257 LITERATURE CITED ........................................................................................... 258 xi Page PUBLICATION 6 (The Bryologist, submitted February 2004): The Buellia aethaleaGroup in the Greater Sonoran Desert Region with some reference to similar species known from North America ................................................................................ 265 METHODS ......................................................................................................... 267 KEY TO THE SPECIES ......................................................................................... 302 DISCUSSION ...................................................................................................... 305 ACKNOWLEDGMENTS ........................................................................................ 311 LITERATURE CITED ........................................................................................... 312 PUBLICATION 7 (The Bryologist, submitted February 2004): The genus Buellia s.l. in the Greater Sonoran Desert Region: saxicolous species with one-septate ascospores containing xanthones ........................................................................................... 316 METHODS ......................................................................................................... 318 SPECIES DESCRIPTIONS ...................................................................................... 318 KEY TO THE SPECIES ......................................................................................... 356 DISCUSSION ...................................................................................................... 357 ACKNOWLEDGMENTS ........................................................................................ 364 LITERATURE CITED ........................................................................................... 364 PUBLICATION 8 (Mycotaxon, submitted March 2004): New and previously not recorded saxicolous species of Buellia s.l. with one-septate ascospores from the Greater Sonoran Desert Region ............................................................................ 368 INTRODUCTION ................................................................................................. 369 METHODS ......................................................................................................... 370 xii Page TAXONOMIC DESCRIPTIONS .............................................................................. 371 KEY .................................................................................................................. 418 ACKNOWLEDGEMENTS ...................................................................................... 429 LITERATURE CITED ........................................................................................... 429 OTHER SPECIES DESCRIPTIONS: Buellia paniformis, B. subdisciformis, B. tesserata and B. uberior ............................................................................................................. 436 TAXONOMIC PERSPECTIVES ................................................................................. 455 REFERENCES ............................................................................................................. 457 APPENDIX I: LIST OF SPECIMENS EXAMINED WITH HPLC ............................ 480 APPENDIX II: CHARACTER CODING FOR THE PHYLOGENETIC ANALYSIS ..... ........................................................................................................................................ 484 BIOGRAPHICAL SKETCH ........................................................................................ 488 xiii LIST OF TABLES Table Page 1. Secondary metabolites in Buellia s.l. ....................................................................... 63 2. Data matrix used for the cladistic analysis .............................................................. 83 3. Thallus characteristics of morphotypes of Buellia dispersa .................................. 113 4. Diagnostic differences in B. amabilis, B. subalbula and B. venusta ..................... 177 5. Diagnostic differences of Buellia christophii, B. prospersa, B. pullata, B. ryanii, B. sequax and B. tergua ......................................................................................... 206 6. Key characters of the Buellia aethalea-group ....................................................... 304 7. References to detailed descriptions of all species known from the Sonoran Desert Region .................................................................................................................... 401 xiv LIST OF FIGURES Figure Page 1. Line drawings of different exciple types I ............................................................... 27 2. Line drawings of different exciple types II .............................................................. 29 3. The ascus type in Buellia s.l. ................................................................................... 35 4. Comparison of ascospore ultrastructure in the Physciaceae .................................... 41 5. Ultrastructure of paraphyses, hyphae and ascospores in the Physciaceae ............... 43 6. Pycnidia, conidiophores and conidia of Buellia s.l. ................................................ 55 7. Distribution of Buellia aethalea, B. argillicola, B. badia, B. christophii, B. concinna and B. dispersa ........................................................................................................ 67 8. Distribution of Buellia eganii, B. halonia, B. lacteoidea, B. lepidastroidea, B. mamillana and B. nashii .......................................................................................... 69 9. Distribution of Buellia navajoensis, B. paniformis, B. prospersa, B. pullata, B. regineae and B. ryanii ............................................................................................. 71 10. Distribution of Buellia sequax, B. sheardii, B. spuria, B. stellulata, B. subaethalea and B. subalbula ...................................................................................................... 73 11. Distribution of Buellia subdisciformis, B. tergua, B. tesserata, B. trachyspora, B. tyrolensis and B. uberior ......................................................................................... 75 12. Distribution of Buellia vilis ..................................................................................... 77 13. Strict consensus tree of one thousand equally most parsimonious trees ................. 89 14. Typical morphotype (morphotype I) of Buellia dispersa ...................................... 115 xv Figure Page 15. Morphotype II and morphotype III of B. dispersa ................................................ 117 16. Light micrographs of thallus, ascospores and ascus .............................................. 119 17. Light, transmission electron and scanning electron micrographs of Buellia dispersa ................................................................................................................................ 121 18. Thallus morphology of Buellia badia .................................................................... 145 19. Anatomy of Buellia badia ..................................................................................... 147 20. Morphology and anatomy of Rhizocarpon saurinum ............................................ 163 21. Buellia subalbula ................................................................................................... 179 22. Buellia amabilis ..................................................................................................... 181 23. Buellia venusta ...................................................................................................... 183 24. Powder X-ray diffraction spectrum of the thallus of B. venusta ........................... 185 25. Buellia christophii and B. pullata .......................................................................... 209 26. Buellia prospersa and B. sequax ........................................................................... 213 27. Buellia ryanii and B. tergua .................................................................................. 225 28. Chasmolithic growth in Buellia sequax on various substrates .............................. 231 29. Variation of epilithic growth in Buellia christophii and B. pullata ....................... 235 30. Ascospore ontogeny of Buellia species with inconspicuous saxicolous thalli from the Sonoran Desert ................................................................................................. 237 31. Range of conidial length in various saxicolous species of Buellia ........................ 241 32. Buellia aethalea ..................................................................................................... 273 33. Buellia eganii ......................................................................................................... 277 34. Buellia lacteoidea .................................................................................................. 283 xvi Figure Page 35. Buellia spuria ........................................................................................................ 289 36. Buellia stellulata .................................................................................................... 297 37. Buellia concinna .................................................................................................... 341 38. Buellia halonia ...................................................................................................... 343 39. Buellia mamillana .................................................................................................. 345 40. Buellia subaethalea ............................................................................................... 347 41. Buellia trachyspora ............................................................................................... 349 42. HPLC chromatograms of Buellia concinna and Buellia halonia .......................... 351 43. HPLC chromatograms of Buellia mamillana and Buellia subaethalea ................ 353 44. HPLC chromatogram of Buellia trachyspora ....................................................... 355 45. Light micrographs of Buellia argillicola ............................................................... 403 46. Buellia lepidastroidea ............................................................................................ 405 47. Buellia nashii ......................................................................................................... 407 48. Buellia navajoensis ................................................................................................ 409 49. Buellia regineae ..................................................................................................... 411 50. Buellia sheardii ...................................................................................................... 413 51. Buellia tyrolensis ................................................................................................... 415 52. Buellia vilis ............................................................................................................ 417 53. Buellia paniformis ................................................................................................. 439 54. Buellia subdisciformis ........................................................................................... 443 55. Buellia tesserata .................................................................................................... 445 56. Buellia uberior ....................................................................................................... 453 xvii INTRODUCTION The genus Buellia De Not. is a large, poorly understood crustose lichen genus in the Physciaceae. Both genus and species concepts are currently subject to much controversy and recent molecular data have even challenged the classical concept of the family (Wedin et al. 2000; Wedin et al. 2002; Helms et al. 2003). Saxicolous species of Buellia with one-septate ascospores are still the least investigated group of the genus. The only concise modern treatment is a publication by Scheidegger (1993), who examined the European species. In North America Imshaug (1951) provided the last extensive treatment of Buellia s.l. Specimens from the Southwest are underrepresented in his dissertation and the Sonoran Desert Lichen Flora Project (Nash et al. 2002) provides an excellent opportunity to study this group in more detail. The region offers a wide range of diverse rock habitats. Many taxa described from the North American Southwest are closely related to European ones and the North American material can thus be compared with European specimens. Lichen taxonomy is still based predominantly on classical data but traditional concepts become more and more challenged, as molecular techniques become applied. Studies of classical data are nevertheless important. Without a thorough understanding of lichen morphology, anatomy, spore ultrastructure and secondary chemistry, molecular information bears little meaning. Classical characters of saxicolous lichens are notoriously difficult to interpret and frequently subject to misinterpretation. With sophisticated technologies it is possible to examine structures more thoroughly. This study investigates the anatomical and 2 morphological characters of a subset of the genus Buellia with advanced microscopy. Important characters like exciple and pycnidium anatomy, and ascus and ascospore structure are examined here in detail. Many saxicolous species are inconspicuous and to some extent hidden within their rock substrate. Thallus characteristics of these species have been examined with techniques developed to study endolithic growth. Lichen secondary metabolites have also been analyzed using standardized thin-layer chromatography (TLC). In addition some specimens were selected and analyzed with high performance liquid chromatography (HPLC). Despite the focus on a select subset of Buellia it is anticipated that this dissertation will have an impact on the taxonomy at both the genus and species level. Based on the results presented here, classical taxonomic concepts of the genus need to be re-evaluated and carefully revised. This dissertation advocates a broad genus concept for species of Buellia s.l. Marbach (2000) recently suggested several new genera for corticolous species. He argued that his reorganization had few consequences for saxicolous taxa. There is little doubt that Buellia s.l. constitutes an amalgam of not necessarily very closely related species. Marbach's (2000) concept that the saxicolous species form a distinct group, is nevertheless problematic. Most segregates of the genus are currently based on very little evidence and should only be accepted if they can be confirmed with molecular tools. The research presented here has not only taxonomic implications. Few studies have used both light and electron microscopy to study lichens. The ultrastructure of the ascus and ascospores of most lichen taxa is still poorly known and spore types are largely based 3 on observations with the light microscope. For example, several modern treatments still refer to ascospores of the Physciaceae as polarilocular even though ultrastructural studies clearly reveal that the spore cells are not divided into separate locules. Ascus types in the Physciaceae are based on studies with the light microscope and little information is available about the corresponding ultrastructure. Finally saxicolous species are usually segregated into epilithic and endolithic, but the studies here present evidence that all saxicolous lichens demonstrate some degree of endosubstratic growth and a more refined concept is discussed. Most results presented in this dissertation have already been published or at least have been submitted for publication. These publications and manuscripts are presented in separate chapters. 4 MATERIAL AND METHODS Specimens examined Representative specimens are listed within publications 1 to 8 and the descriptions of additional species. Detailed information for all ASU specimens is available online at http://seinet.asu.edu/collections/selection.jsp. Specimens on loan from the following herbaria were also examined and annotated: B, BM, C, CANL, CAS, COLO, FH, G, GZU, H, hb. Kalb, hb. Knudsen, hb. Lendemer, hb. Nordin, hb. Printzen, hb. Scheidegger, hb. van den Boom, L, M, MEXU, MICH, MIN, MSC, MU, OMA, PH, S, SASK, SBBG, STU, TUR, UC, UPS, US, W. Methods LIGHT MICROSCOPY All specimens were examined with light microscopy using hand- and cryosections. The cryosections were prepared in frozen water on a Reichert-Jung HistoStat 855. Specimens were examined on a Leica/Wild M420 dissecting microscope and a Leica DMLB compound microscope. Conventional bright field microscopy (BF), phase contrast (PhC) and differential interference contrast (DIC) were used. Asci stained with 0.05% aqueous Calcoflour White were examined in a Leica TCS NT Scanning Confocal Laser Microscope in the ASU Keck Laboratory. 5 SCANNING ELECTRON MICROSCOPY Specimens were prepared according to Bungartz et al. (2002, Publication 1) and observed in a Jeol JSM-840A scanning microscope at accelerating voltages between 7 and 12 kV TRANSMISSION ELECTRON MICROSCOPY Specimens were prepared according to Bungartz et al. (2002, Publication 1). To improve dehydration and infiltration, this protocol has been modified according to Bungartz & Nash (2004a, Publication 4). Specimens were observed in a Philips EM 201 transmission electron microscope and a Philips CM 12 scanning transmission electron microscope at accelerating voltages of 60 and 80 kV SPOT TESTS All specimens were spot tested using conventional spot test reagents (Nash III et al. 2002). THIN-LAYER CHROMATOGRAPHY All specimens were routinely examined with standardized thin-layer chromatography (Culberson & Kristinsson 1970; Culberson & Johnson 1982; White & James 1985; Orange et al. 2001). TLC-plates were interpreted with the computer program WINTABOLITES (Mietzsch et al. 1994), and scanned for permanent record (Egan 2001). 6 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY A subset of the specimens analyzed with TLC were more closely examined by Dr. John A. Elix (Australian National University, Canberra) with standardized high performance liquid chromatography (HPLC, Elix et al. 2003). APPENDIX I is a list of all specimens examined with HPLC. CLADISTICS Parsimony Analysis Parsimony analyses were conducted using the program PAUP* version 4.0b10 (Swofford 2002). The morphological character dataset is presented in table 2; a description of each character and the coding scheme is included in APPENDIX I. Most characters were the same as the ones used for a similar analysis described by Nordin (2000). The Sonoran species and a few corticolous taxa with one-septate spores were added to evaluate their taxonomic position and to re-examine Nordin's (2000) conclusions. As in Nordin’s previous analysis all characters were equally weighted and multistate characters were not ordered. Tree searches were conducted using heuristic search options that included random addition sequences (1000 replicates) holding 1 trees per replicate, followed by tree-bisection-reconnection (TBR) branch swapping, with retention of multiple parsimonious trees (MAXTREES = 1000). The most parsimonious trees were rooted using Rinodina as the outgroup, and summarized as strict, semistrict and 50% majority rule consensus trees. Non-parametric bootstrapping (Felsenstein 1985) was used to evaluate clade support; bootstrap proportions were estimated from 1000 7 bootstrap replicates incorporating the same heuristic parsimony search options as using the standard parsimony analysis. Bremer support was not evaluated because it only measures stability as judged by tree length for a single data set and the support values would thus not have been directly comparable to the original data set examined by Nordin (2000). Character Coding To include saxicolous taxa from the Sonoran Desert Region a few modifications to Nordin's (2000) original character set were necessary. The coding of polymorphisms was avoided. A character, which is present only in a subset of a species, is not necessarily a polymorphism. True polymorphisms are characters, which are exclusive, but both occur in two different populations of the same taxon. For example, a petal of a flower cannot be yellow and red at the same time, it is either yellow or red. Nevertheless it is conceivable that the same plant species consists of populations with either yellow or red flowers, even a single plant could have flowers, which have red or yellow petals. This situation is different for a species, which has the capability to synthesize red petal color, but nevertheless has some white petals because these are lacking the red pigment. Analogous to this example, it does not seem justified to code the presence or absence of a secondary lichen metabolite as a polymorphic character. Nordin (2000) suggested that B. alboatra was polymorphic, because both chemotypes with or without norstictic acid can be found. However, there may be several reasons why some populations of B. alboatra do not synthesize norstictic acid. It cannot necessarily be assumed that these specimens entirely lack the ability to produce the secondary metabolite. The fact that 8 other populations are capable of synthesizing norstictic acid is evidence that the character should not be coded as polymorphic. Nordin (2000) also coded some anatomical characters as polymorphic. For example, some populations of B. pharcidia apparently have spores without longisepta but others form one or two longisepta. It is not conceivable why some specimens do not form longisepta, but longisepta are present at least in some of the specimens. The species should therefore not be coded as polymorphic, but as a species which can produce up to two longisepta. Most other instances, which Nordin (2000) coded as polymorphisms, represent similar situations. Characters, which are represented by a multitude of polymorphisms, may not be suitable for the analysis and might better be excluded. For some of the species little or no information was available and character states were then coded as “?”, i.e. not known. In other instances data were insufficient for a character to be included. Therefore, the following characters used by Nordin (2000) were not taken into account: Width of paraphysal tips (character no. 15 in Nordin 2000).—Paraphyses in immature apothecia are often very narrow. The width also varies among paraphyses of the same apothecium. Finally the amount of carbonization also contributes to this variability. Strongly pigmented cells usually appear more distinctly swollen than moderately pigmented cells. As a result of this variability, Nordin (2000) coded three species as polymorphic. The character was excluded from the analysis because it was deemed too variable. 9 Carbonized cells in the exciple (character no. 11 in Nordin 2000).— The amount of pigmentation and thus carbonization of the exciple cells is also variable. Generally the outermost exciple cells are more heavily pigmented than inner exciple cells. Cells are often entirely carbonized in mature or overmature apothecia. Nordin (2000) suggested that only cells which are entirely pigmented should be referred to as carbonized. This definition is problematic because the pigmentation of cells is a gradual process and there is no distinct way to decide when a cell is completely carbonized. The character was therefore not included in the analysis. Secondary lichen metabolites in the exciple (character no. 12 in Nordin 2000).— Thallus and exciple chemistry were not examined separately for most of saxicolous specimens. Too little information was available to include this character. Absence or presence of a brown K+ yellow pigment within the exciple (character no. 38 in Nordin 2000)—A yellow color reaction caused by secondary metabolites cannot reliably be distinguished from a similar reaction caused by non-acetone soluble pigments. The character was therefore not included in the analysis. Enclosure of young apothecia (character no. 4 in Nordin 2000)—The character was not examined in all species and data, if included, would therefore not have been reliable. Nine characters were added to Nordin's (2000) analysis. A detailed list of all characters used in the analysis is included as APPENDIX II. 10 TAXONOMIC CONCEPTS Taxonomic Concept of Buellia within the Physciaceae The family Physciaceae was first introduced by Zahlbruckner (1898) for foliose and fruticose species. Zahlbruckner (1926) included crustose genera, like Buellia and Rinodina, in the Buelliaceae. Poelt (1973) unified the two families. This concept remained unchanged, until molecular data demonstrated that the Caliciales could not be maintained as a distinct order but instead nested within the Physciaceae (Wedin et al. 2000; Wedin et al. 2002). The family name Caliciaceae is older than Physciaceae and thus has taxonomic priority. However, Caliciaceae traditionally refers to a well confined group of species characterized by prototunicate asci disintegrating into a mazaedium whereas Physciaceae include more diverse taxa. Wedin & Grube (2002) therefore suggested conserving Physciaceae against Caliciaceae and to include the calicioid species within an emended concept of the Physciaceae. The Physciaceae sensu Wedin & Grube (2002) would thus be characterized by two large clades: (1) the Physcia- (or Rinodina) clade including all foliose and fruticose taxa and Rinodina s.l., and (2) the Buellia-clade with genera closely related to Buellia s.l. and the calicioid taxa. Helms (2003) argued that this obvious division into two major clades should preferably be used to distinguish two families. Instead of including the Caliciaceae within the Physciaceae, Helms, Rambold and Friedl in Helms (2003) therefore emended the family concept for both Physciaceae and Caliciaceae. Their concept divides the traditional family Physciaceae excluding Buellia and related taxa, which are transferred 11 into the Caliciaceae. Accordingly, the Physciaceae sensu Helms (2003) are more narrowly confined, including only Rinodina and related foliose and fruticose taxa. Unfortunately the classical characters used to define the two emended families do not correlate as clearly with the two clades as Helms (2003) suggests. In his table 3.5 (p. 29, Helms 2003) he lists 11 taxa with characters deviating from the emended family concepts. Several species of Buellia s.l. from the Sonoran Desert Region could be included within this list. They have distinct wall thickenings and at least two species (Buellia aethalea, B. lacteoidea) have a hypothecium which is not consistently pigmented. Nevertheless, Helms (2003) argued that only Australienea seriously conflicts with his concept because it is the only species in the Buellia clade with a Lecanora-type ascus. Several species in Buellia s.l. have an unusually wide central tholus not staining in iodine, e.g., B. oidalea or B. oidaliella (Nordin 2000) and B. badia. These species, nevertheless, all have an ascus with a distinct, narrow, I+ blue band at the ascus apex and it can therefore be argued that these asci thus belong to the Bacidia-type, even though they superficially resemble the Lecanora-type. Genera like Pyxine or Dirinaria, which have a Bacidia-type ascus, were generally believed to be closely related to foliose genera of the Rinodina-clade (Nordin & Mattsson 2001). Helms (2003), however, demonstrated that these genera are more closely related to Buellia s.l. The emended family concept by Helms (2003) has formal nomenclatural advantages that avoid the conservation of one family name against another one. However, it does not address the concern of Wedin & Grube (2002) that Caliciaceae traditionally refers to a much more narrowly confined group. As an alternative Helms (2003) therefore suggested 12 resurrecting Zahlbruckner's (1926) Buelliaceae and conserving that name against Caliciaceae. At this stage it appears to be a matter of personal preference which family concept is adapted: two newly emended families Caliciaceae and Physciaceae sensu Helms (2003) or a single conserved family Physciaceae sensu Wedin & Grube (2002). In this dissertation the concept of Wedin & Grube (2002) is adopted and the “Caliciaceae” are thus referred to as calicioid Physciaceae. Taxonomic Concept of the Genus Buellia De Not. The lichen genus Buellia was described by De Notaris (1846) to accommodate crustose species with one-septate, brown ascospores and black apothecia. De Notaris named the genus in honor of his friend Esuperanzo Buelli (Leunis 1877). The name soon became accepted by early lichenologists (Massalongo 1852; Körber 1855; Fries 1860) and many new species were subsequently assigned to the genus. Flotow (1849b) segregated Diplotomma from Buellia based on a “false” thalline exciple. Massalongo (1852) refined this concept adding the pluriseptate spores as the major character of the genus. Fries (1861), however, emended the genus concept of Buellia to include species with three-septate ascospores. As a consequence, Diplotomma was not universally accepted, and species with pluriseptate spores were either described as Buellia or Diplotomma. Nordin (1996; 2000) provided an excellent account of the species with pluriseptate spores and demonstrated convincingly why the majority of species with pluriseptate spores must not be included in Diplotomma s.str. Zahlbruckner (1926) was the first author to confine Buellia to species with lecideine apothecia, and thus distinguished the genus from Rinodina. This concept was essentially 13 used with very little refinement until the 1980’s. Hafellner et al. (1979) emphasized the thin walled spores of Buellia as an additional character segregating the genus from Rinodina, until Scheidegger (1993) discovered species with ± distinct thickenings of the spore septum. Kalb (1986) introduced the genus Hafellia, to segregate species with an oil inspersed hymenium and lateral spore wall thickenings. Scheidegger (1993) validated the genus Amandinea, a genus proposed by Choisy (1950) to accommodate species with filiform conidia. Among lichen taxonomists there is currently very little agreement about which segregates of Buellia must be accepted as distinct genera, and a widely accepted species concept of the genus does not currently exist. Most recently Marbach (2000) published an extensive account on corticolous species from the tropics and subtropics, and suggested that none of these species belong to Buellia s.str. Instead Marbach (2000) accepted thirteen genera, nine newly described in his treatment. In contrast, Nordin (2000) did not adopt Marbach's (2000) approach. Nordin (2000) argued that “….Segregation of new taxa should not be taken too lightly…” (p. 10, Nordin 2000) and included even species with pluriseptate spores within Buellia s.l. Several reasons are responsible for these vastly different approaches: (1) The nomenclature of Buellia and its formal typification remains presently unresolved. The current taxonomic code (Greuter et al. 2000) lists Buellia de Not. as a conserved genus with the type Buellia disciformis, but a proposal by Moberg et al. (1999) to change this listed type to Buellia aethalea is pending. 14 (2) No monograph for all taxa exists. Various modern treatments have been confined to particular subsets, like geographic groups (Imshaug 1951; Scheidegger 1993; Marbach 2000), saxicolous vs. corticolous species (Scheidegger 1993; Marbach 2000) or species with pluriseptate vs. one-septate spores (Scheidegger 1993; Nordin 2000). (3) Data for classical characters such as morphology, anatomy, chemistry and spore ultrastructure, are still fragmentary for the majority of taxa. Especially saxicolous species with one-septate ascospores are still largely unknown, and this dissertation is a first attempt to provide a detailed analysis of these taxa. (4) The sheer number of species originally described as Buellia is daunting. The Index Fungorum (http://www.indexfungorum.org/) lists more than 4000 species names! Most names are clearly synonyms of species described multiple times. Reasonable estimates suggest that ca. 400 species belong to the genus if a generous genus concept is adopted (Hawksworth et al. 1995; Kirk et al. 2001). (5) Virtually no attempts have been made to evaluate classical data with molecular tools. Molecular studies confirm two major clades of the Physciaceae (Wedin et al. 2000; Wedin et al. 2002; Wedin & Grube 2002; Helms 2003; Helms et al. 2003). Nevertheless, all genera are currently based only on classical data (e.g. Kalb 1986; Scheidegger 1993; Marbach 2000). It has been a common experience in lichen taxonomy that classical groups have to be revised drastically based on molecular evidence. For example, the entire order Caliciales was found to be a clade nested within the Physciaceae (Wedin et al. 2000; Wedin et al. 2002; Wedin & Grube 2002). Classical data, therefore, must be carefully evaluated with molecular tools. Only a combined approach will help to resolve 15 the phylogeny of the group. New genera should only be accepted, if they are well supported both by classical and molecular evidence. Genera should be recognized as evolutionary units of closely related species based on their phylogeny. Segregating new genera solely on the basis of a few classical characters is not a desirable approach to modern taxonomy. A broad concept of Buellia s.l. is adopted in this dissertation even though many species are not necessarily closely related. The genus Buellia s.l., as accepted here, can be characterized by its (1) crustose growth, (2) predominantly lecideine apothecia with a dark pigmented hypothecium, (3) Bacidia-type ascus, and (4) one-septate to pluriseptate ascospores with or without moderate wall and septum thickenings. 16 MORPHOLOGY AND ANATOMY Thallus Morphology Major differences in thallus morphology are the result of a different thallus ontogeny. Thalli either develop from a continuous crust or as separate areoles (Sheard 1964). Continuous thalli soon become fissured. These fissures are probably a result of drying and re-wetting. The thalli are rimose if these fissures remain few and rarely merge with one another (e.g., B. subalbula). Some species are rimose areolate but their “false areoles” are derived secondarily from fissures because their edges are not lined with a cortex (e.g., B. mamillana or B. trachyspora). In contrast, distinctly areolate species develop distinctly separate areoles, which are often widely dispersed and frequently become subsquamulose (e.g., B. dispersa or B. nashii) or even squamulose (e.g., B. badia). These areoles are usually lined by a ± intact cortex even if they are growing closely together. Sometimes true areoles become fissured and in these instances it is difficult to distinguish rimose-areolate from areolate thalli. Most thalli establish on a non-lichenized weft of fungal hyphae referred to as prothallus. Hyphae from the prothallus are characteristically strongly pigmented and thus appear black. The pigmentation is usually stronger towards the thallus margin. Thus, a prothallus can be distinguished at least as a dark outline delimiting separate thalli from one another. If the prothallus becomes more extensively developed, extending below and between the areoles, it may be referred to as a hypothallus. In most species with a distinct hypothallus like B. stellulata or B. spuria, hyphae aggregate closely and single strands of hyphae cannot be distinguished macroscopically. In contrast, the prothallus 17 typically developing in B. tesserata appears fimbriate, i.e. composed of thin, radiating hyphae. In B. ryanii, hyphae are bundled into distinct, arachnoid strands. Several species, which develop from independent areoles, have no distinct prothallus, e.g. B. dispersa, B. lepidastroidea, B. nashii and B. paniformis. Other species, which are characterized by a prothallus, nevertheless are areolate (e.g., B. ryanii). The thallus morphology of crustose lichens thus can be quite variable, and it is sometimes argued that morphological characters have little taxonomic significance because they are subject to environmental modification (e.g., Weber 1968; Weber 1977). Evaluating thallus morphology in the context of environmental modification is very difficult, because complex environmental factors interact with the thallus (Hawksworth 1973). Specimens of Buellia s.l. on hard, siliceous substrates surfaces are usually distinctly epilithic, frequently have a distinct prothallus and usually do not contain calcium oxalates. In contrast, specimens on soft, porous or slightly calcareous substrates usually show some degree of substrate penetration and are often filled with large amounts of calcium oxalate or other mineral crystals. Thalli growing on smooth, hard surfaces are generally more continuous than on porous substrates with a coarse crystalline surface. Shaded forms are generally paler than their exposed counterparts. Sheltered specimens have a smooth surface. If specimens grow in exposed microhabitats their surface becomes eroded. This variation can be interpreted in two ways: (1) modifications of the thallus morphology caused by the environment, or (2) as a specific adaptation to a distinct microhabitat. Species with a wide ecological amplitude should display more thallus 18 variation than species with a narrow amplitude. If species consistently show the same anatomical, chemical and ultrastructural characters but display large variation in thallus morphology, it can be argued that this variation is a result of environmental modification and not adaptation. Buellia dispersa is a good example of a species, which best illustrates this extreme (Bungartz et al. 2002, Publication 1). Thallus variation observed in this species is enormous, but all specimens have very consistent anatomical and chemical characters, and the different morphotypes have therefore been treated as a single species (Bungartz et al. 2002, Publication 1). However, preliminary molecular data suggest that some of the morphotypes may have to be recognized as separate species. Thus the variation may be less extreme than initially assumed. At least B. nashii has been recognized here as a separate species closely related to B. dispersa but with a different chemistry, exciple pigmentation and distribution (Bungartz 2004, Publication 8). Some species display very little morphological variation. Buellia subalbula, for example, is restricted to coastal, ± maritime and calcareous substrates. The large amounts of calcium oxalates in the thallus are not merely the result of environmental modification, as the species only grows on calcareous rocks. Even species, which can be found in several different habitats, do not necessarily display much thallus variation. These species maintain a similar morphology, even though they are common, widely distributed and usually grow in a variety of different microhabitats. They are usually easily recognized by morphological characters. The yellowish green, distinctly areolate B. halonia is a good example. The species is confined to coastal habitats but grows in sheltered as well as exposed, shaded as well as sunny 19 habitats. Even though it can be found across these different microhabitats, it is always easily recognized by its thallus morphology. In the past species with inconspicuous, thin and dark thalli were often confused and lumped for convenience. It can, however, be demonstrated that even these inconspicuous species are distinguished by distinct and diagnostic characters (Bungartz et al. 2004b, Publication 5). Thallus Anatomy The genus Buellia shows little variation in thallus anatomy. All species are phenocorticate, i.e. they have no cellular cortex. Instead hyphae extending from the medulla and photobiont layer are loosely aggregated and disintegrate towards the thallus surface. A residue of dead hyphae forms an epinecral layer on the surface. In some species, a false pruina can be observed from flaky, disintegrating hyphae, but in most species with pruina crystals of calcium oxalate or secondary metabolites (e.g., xanthones) are interspersed with the false pruina. Species with a smooth surface are usually characterized by a strong layer of mucilage binding together the epinecral layer. Such a species is B. paniformis, where even the immature apothecia are usually covered by an epinecral layer, which extends across the surface (Fig. 53D). Thallus pigmentation is a result of the pigment present on the cortex, the thickness of the epinecral layer and the thickness of the photobiont layer below. Buellia dispersa and B. nashii show extensive variation. Their thalli are pale to deep brown, if they have a thin epinecral layer, a distinctly pigmented phenocortex and a moderately thickened photobiont layer. Specimens with a thick epinecral layer are much paler or even whitish, 20 and specimens with a thick photobiont layer are dark gray or olive gray (Bungartz et al. 2002, Publication 1). Apothecia Morphology and anatomy of the apothecium provides some of the most important diagnostic characters of Buellia s.l. The ascus type, spore septation, ontogeny and ultrastructure, the hypothecium pigmentation, the hymenium inspersion and the exciple structure have all been used to distinguish different species as well as to segregate new genera. These characters are thus also important for the taxonomic concept of the Physciaceae. ANATOMY OF THE EXCIPLE AND HYPOTHECIUM Zahlbruckner (1926) emended the genus concept of Buellia, emphasizing the lecideine apothecia as the main diagnostic character of the genus. Scheidegger (1993), however, referred to apothecia of Buellia as cryptolecanorine, zeorine, biatorine or even lecanorine. This concept is problematic in several respects: Immersed apothecia are not necessarily lecanorine if they have a distinct, proper exciple but no indication of a thalline margin. The term “zeorine” is obsolete, because it refers to apothecia with both a proper and a thalline exciple. All lecanorine apothecia have some reduced inner hyphae, which represent a reduced proper exciple, thus “zeorine” apothecia are anatomically identical with lecanorine apothecia. The term biatorine refers to apothecia with an exciple, which is not carbonized throughout. All species of Buellia s.l. have a proper exciple, and at least the outermost cells of the exciple 21 as well as paraphysal end cells are usually strongly carbonized by a brown pigment cap (Fig. 5A,B). There is no clear distinction between pigmentation and carbonization. Carbonized cells are strongly pigmented cells. An exciple entirely composed of strongly pigmented cells is more brittle (“carbonlike”) than plectenchyma with less pigmented cells. The amount of carbonization is generally quite variable in most species. Nordin (2000) suggests that only cells, which are completely pigmented throughout, are carbonized. This concept is too narrow to accommodate carbonization as a dynamic process beginning with pigmentation of the cell walls and ending with a cell completely filled with pigment (Fig. 5). Very rarely exciple hyphae are completely and solidly carbonized throughout, and these cells are probably dead (e.g., the outer exciple of B. trachyspora, Bungartz et al. 2004c, Publication 7). However, most exciple cells probably continue to grow, although they have a pigment cap. Especially young apothecia are usually less strongly pigmented and not brittle. These apothecia could be referred to as biatorine. The term is not used here, because truly biatorine apothecia are usually faintly pigmented even with age. In several species of Buellia s.l. part of the thallus remains attached to the proper exciple for considerable time after apothecia emerge from the thallus. These thallus remains must not be confused with a distinct thalline exciple, and the apothecia should not be referred to as lecanorine. In Buellia halonia, large necrotic fragments of thallus material often remain attached to the exciple (Bungartz et al. 2004c, Publication 7). In Buellia stellulata or B. spuria the young exciple is characteristically covered by a thin, epinecral layer, here referred to as thalline veil. Buellia alboatra and B. venusta regularly 22 have a thalline collar, i.e. an irregular ring of undifferentiated thalline material attached to the exciple (also see Nordin 2000). The only species which clearly has a thalline exciple is B. mamillana. However, the thalline exciple soon becomes excluded in this species, because the inner, proper exciple expands and pushes the thalline exciple below the apothecium (Bungartz et al. 2004c, Publication 7). Scheidegger (1993) described several distinct exciple types but his concepts were not adopted generally by other lichenologists, because the distinction of the different types relies on cumbersome examination of the hyphal arrangement throughout the exciple. Only thin microtome sections allow this differentiation of the exciple structure to be observed. In thick sections hyphae are obscured by the strong pigmentation. In contrast to Scheidegger (1993), Marbach (2000) and Nordin (2000) both emphasized pigmentation of the exciple and did not describe the structure in detail. Their different types were based on pigmentation, but nevertheless largely correspond to Scheidegger’s (1993) structural types. Scheidegger (1993) also argued that certain exciple pigments were characteristic for a particular exciple structure. He distinguished five different pigments (pigment A–E) based on their color and reactions with 10% KOH and concentrated HNO3. Meyer & Printzen (2000) introduced a standardized nomenclature of exciple pigments based on a series of color tests applied: (1) H2O → KOH (10-20%) → HCl (10-20%), and (2) H2O → HNO3 (50%) → KOH (10-20%) → HCl (10-20%). Most of Scheidegger’s (1993) pigments easily translate into the nomenclature introduced by Meyer & Printzen (2000), but not all pigments described by Scheidegger (1993) and Meyer & Printzen (2000) can be confirmed here. Some color reactions are misleading because the pigments do not 23 occur as pure colors, but are “superimposed”, i.e. several pigments are usually mixed in different concentrations. In addition, some colorless lichen substances like atranorin and norstictic acid react with KOH and further obscure the observations. Pigment A is equivalent to cinereorufa-green, an aeruginose, distinctly HNO3+reddish violet pigment. The pigment is not restricted to the aethalea-type, but also occurs in the leptocline-type (Buellia halonia) and the dispersa-type (Buellia nashii). The pigment always occurs on the cell surface as a diffuse pigment and eventually dissolves with repeated application of test reagents. Species which are characterized by the presence of this exciple pigment, also have pycnidial ostioles colored with this pigment. The pigment is easily recognized, if it occurs in strong concentrations throughout the entire exciple and across the epihymenium. In Buellia nashii and Buellia lepidastra (and to some extend in B. sheardii) it is, however, restricted to the outermost exciple cells. The pigment also frequently occurs in low concentrations and is then overshadowed by the presence of brown pigments in the cell wall. Pigment B is probably identical with elachista-brown. This dull brown pigment changes little in color, if the tests of Meyer & Printzen (2000) are applied. It is characteristic for outer exciple cells and the paraphysal caps of all species and occurs inside the cell wall (Fig. 5A,B). Only Buellia vilis does not have this pigment. Pigment C is identical with atra-red and occurs only in Buellia vilis. It appears to be located in the cell wall, but washes out with repeated applications of HNO3. In addition to the unique pigment, Buellia vilis can be further characterized by a structurally unique exciple (see vilis-type below). 24 Pigment D is a deeper, more reddish brown pigment identical with leptoclinoidesbrown. It is typically restricted to cell walls of the hypothecium and occurs in most species. In Buellia aethalea the pigment can be present in very low concentrations, and the hypothecium may thus be very pale brown or even hyaline. The pigment leptoclinoides-brown is particularly characteristic for the dispersa-type, where it extends far into the inner exciple of mature apothecia. The color reactions of pigment E are identical with leptocline-brown. Meyer & Printzen (2000) based this pigment on Buellia leptocline. The reference collection in M (Lich. Helv. Exs. 311, Schaerer & Hepp 839., M-0061325) shows a distinctly fleeting K+ yellow color reaction, but the reaction with HCl is not reddish brown. Atranorin is present in the exciple of this collection, and it is not possible to distinguish the K+ yellow reaction caused by atranorin from a reaction possibly caused by a separate brown pigment. Several other reports of pigment E (leptocline-brown) may also be based on the presence of colorless, K+ yellow lichen metabolites like atranorin or norstictic acid within a brown exciple. Nordin (2000), however, emphasized that he was able to observe a K+ yellow reaction even in species, which did not contain any secondary metabolites in their exciple. Both Nordin (2000) and Marbach (2000) mention additional pigments, especially yellow or reddish orange anthraquinones, which have not been found in any saxicolous specimens examined here. 25 The aethalea-type exciple The aethalea-type is the most common exciple-type in Buellia s.l. It is characterized by thin, more or less parallel hyphae, similar in orientation and structure to the paraphyses (Fig. 1 A,B). The proper exciple of an apothecium is essentially the same layer as the hypothecium (the inner envelope) and all excipular hyphae are thus connected with the hypothecium. This connection is often not very distinct in the aethalea-type, because the excipular hyphae have the same upright orientation as the paraphyses, and thus often appear to form a separate layer outside the hypothecium. In species with apothecia remaining immersed in the thallus, the aethalea-type exciple is more strongly reduced to a few, narrow hyphae (e.g. Buellia aethalea, Buellia eganii, Buellia lacteoidea, Bungartz & Nash 2004c, Publication 6). The inner excipular hyphae of the aethalea-type are leptodermatous to mesodermatous, narrow and parallel and thus correspond to textura oblita. The outermost cells are typically swollen and pigmented by a brown pigment cap. They are structurally similar to the inner exciple and most appropriately also referred to as textura oblita. In species with distinctly sessile apothecia, the outer part often becomes more prominent with age and the outer cells then appear distinctly swollen (Fig. 1B). Scheidegger (1993) referred to these swollen outer cells as textura angularis or prismatica. Buellia christophii is a good example of a species with an aethalea-type exciple and a distinctly thickened outer layer. Scheidegger (1993) suggests that the aethalea-type is generally characterized by cinereorufa-green, but nevertheless includes many species which lack that diffuse, 26 aeruginose pigment. Only the outermost cells of the aethalea-type are carbonized and the type thus corresponds ± with Nordin's (2000) description of an exciple, which is distinctly darker in the outermost part. It also corresponds to some extent with Marbach's (2000) type 2. It is not, however, structurally identical with the exciple of Endohyalina circumpallida (Marbach 2000). → FIGURE 1. Line drawings of different exciple types I.—A. The aethalea-type (immersed apothecium of Buellia aethalea).—B. The aethalea-type (sessile apothecium of Buellia spuria).—C. The dispersa-type (Buellia nashii).—D. The leptocline-type (Buellia halonia). 27 28 → FIGURE 2. Line drawings of different exciple types II.—A. The mamillana-type (immature apothecium of Buellia mamillana with distinct thalline exciple).—B. The mamillana-type (later stage of the apothecium ontogeny of B. mamillana with expanding proper exciple).—C. The trachyspora-type (Buellia trachyspora).—D. The vilis-type (Buellia vilis). 29 30 The dispersa-type exciple The dispersa-type can be differentiated into three distinct zones (Fig. 1C). The innermost hyphae radiate from the hypothecium towards the outer exciple. In young apothecia the deep reddish brown pigmentation does not extend very far into the central exciple, but with age the exciple becomes more evenly pigmented throughout. The inner hyphae of the central exciple are mesodermatous, ± parallel (textura oblita) and structurally similar to hyphae of the hypothecium. The outermost exciple cells are distinctly swollen, shorter and more isodiametric and have been referred to by Scheidegger (1993) as textura angularis even though they are not distinctly paraplectenchymatous. This exciple ± corresponds to Nordin's (2000) description of an exciple, which has a distinctly paler middle part. Some species, which Marbach (2000) assigned to his type 2, may also belong here. The leptocline-type exciple The leptocline-type is evenly pigmented throughout by a dull brown pigment (Fig. 1D). In B. halonia, the aeruginose pigment cinereorufa-green is also present and the exciple thus appears fuscous brown to distinctly aeruginose. Several species assigned to this type by Scheidegger (1993) have atranorin within their exciple, which is most likely the cause for the KOH+ yellow diffusing color reaction. This reaction is not necessarily indicative for a separate brown pigment (see comments about pigment E, leptoclinebrown). Nordin (2000) also recognizes this evenly pigmented exciple, but does not describe the exciple structure in detail. 31 The hyphae of the leptocline-type are little differentiated throughout the exciple. They are mesodermatous with a distinctly pigmented wall, initially ± parallel but with age become more and more intricately interwoven (i.e. they form a dense textura intricata). The outer cells do not form a distinctly separate zone from the inner exciple. Scheidegger (1993) refers to them as textura oblita because they sometimes appear less interwoven than hyphae of the inner exciple. The mamillana-type exciple The exciple of Buellia mamillana does not correspond with any other species (Bungartz et al. 2004c, Publication 7). It remains pale during most of the ontogeny of the apothecium (Fig. 2A,B). In young apothecia hyphae from the proper exciple are almost absent and a distinct thalline margin is developed (Fig. 2A). Emerging from the thallus this thalline margin is increasingly replaced by expanding hyphae from the proper exciple (Fig. 2B). The pigmentation of the outermost cells is caused by a dull brown pigment, which is also present in the paraphysal caps of the epihymenium. This pigmentation is probably identical with the dull brown pigment cap of other Buellia species but the pigment is less concentrated, and the disc and exciple of young apothecia thus appear dark brown rather than black. The trachyspora-type exciple The trachyspora-type (Fig. 2C) is characteristic for Buellia trachyspora, a tropical to subtropical species (Bungartz et al. 2004c, Publication 7). It is characterized by a strongly blackened (carbonized) outer exciple of small, globular cells (textura globularis) and a 32 reddish brown, large-celled and distinctly paraplectenchymatous inner exciple (textura angularis). The inner exciple pigment is ± identical to leptoclinoides-brown. The outer hyphae are blackened as a result of an extremely strong pigment concentration throughout the small globular cells. The pigment shows the same color reactions as pigment B (cf. elachista-brown). The vilis-type exciple The vilis-type is unique (Bungartz 2004, Publication 8). It is only known from Buellia vilis (Fig. 2D). This species has a hyaline hypothecium of thin strongly interwoven hyphae which extend throughout the inner exciple and react very strongly iodine-blue. The outermost exciple cells are very deeply pigmented, i.e. strongly carbonized, and can barely be distinguished from one another. The blackish red pigment reacts deep purple with HNO3, a very diagnostic reaction (distinctly different from the HNO3+ reddish violet reaction of cinereorufa green). Asci and Ascospores The current concept of the Physciaceae (Eriksson et al. 2003) is unified by ascus anatomy and spore structure. It can be demonstrated that an emended concept of the Physciaceae s.l. (Wedin & Grube 2002) including the former Caliciales is well supported by the ultrastructure of the spores. ASCUS TYPE The crustose lichen genera Buellia s.l. and Rinodina s.l. were treated by Zahlbruckner 1926) within the Buelliaceae. Poelt (1973), Henssen & Jahns (1974) and Hafellner (1979) 33 included the Buelliaceae in the Physciaceae, reflecting a general shift of lichen taxonomy to emphasize microscopic anatomical characters as more reliable than variable morphological features. Luttrell's (1951; 1955) classification of the ascomycetes based on apical ascus structures was highly influential, and the unification of the Buelliaceae with the Physciaceae by Poelt (1973) was largely based on the identical ascus type of the crustose, foliose and fruticose genera. The ascus type of the Physciaceae is a typical lecanoralean ascus, i.e. it is functionally arrested-bitunicate with a semi-fissitunicate dehiscence. The majority of lichenized fungi have this ascus type in common (Lecanoromycetidae sensu P.M. Kirk, P.F. Cannon & J.C. David; Lecanoromycetes sensu Myconet). Recent molecular studies suggest that prototunicate asci independently derived several times, probably as a reduction of the lecanoralean ascus (Helms 2003; Schultz 2003). Wedin et al. (2000) therefore no longer accepted the “Caliciales” and suggested the inclusion of the “Caliciaceae” within the Physciaceae. The “Caliciaceae” have the same spore type as the Physciaceae, but their prototunicate asci soon disintegrate into a mazaedium (Fig. 4A,B). Within the Physciaceae, two major clades have been recognized: the Rinodina (or Physcia)-clade (Clade A) and the Buellia-clade (Clade B) (Grube & Arup 2001; Wedin et al. 2002; Helms 2003; Helms et al. 2003). Ascus structure is one of the most important 34 → FIGURE 3: The ascus type in Buellia s.l. (A-C. Light micrographs, D.–E. Transmission electron micrographs).—A. Typical Bacidia-type ascus stained with Lugol’s iodine (B. sheardii, Nash 10086, ASU–holotype).—B. Modified Bacidia-type ascus with a wide, unstained inner cone (c) with parallel I+ blue flanks (f) and a narrow iodine+ blue band (arrow) at the ascus apex (Buellia badia, Arnold, Lich Esc. no. 1505, UPS–neotype).—C. Light micrograph of the Bacidia-type ascus stained with calcoflour and observed in the scanning confocal microscope: the spore walls show a bright fluorescence, but the tholus is dark throughout (B. tesserata, Marsh 7125, ASU).—D. Immature ascus of B. spuria (Nash 35879, ASU). E. Ascus apex of Buellia capitis-regum (Bungartz 1854, hb. Bungartz; for designation of the different layers see Bellemère 1994): The a- and b-layer (ab) are barely visible and cannot reliably be distinguished (possibly as a result of fixation artefacts); (c) outer electron opaque c-layer; (d1) d1-layer, i.e. the outer tholus, which is distinctly laminated (in light microscopy this outer part stains deep blue with Lugol’s iodine); (d2) d2-layer, i.e. the inner tholus, which is not layered and ± homogeneous (not staining in Lugol’s iodine); (oc) ocular chamber. 35 36 characters segregating the two clades (Helms 2003). The two major types in the Physciaceae are predominantly based on iodine reactions of the ascus apex but correspond well with the ascus ultrastructure. Marbach (2000) suggested that the variation in iodine staining had little diagnostic relevance and combined both as the Physciaceae-type ascus. This concept should not be adopted, because it ignores valuable information. The Rinodina-clade has a Lecanora-type ascus with a central cone not staining in Lugol’s iodine and two broad I+ blue tholus flanks. The central, I– cone extends throughout the entire tholus and sometimes even expands towards the ascus tip. In contrast, the Buellia-clade has a Bacidia-type ascus with a parallel or ± converging, I– cone not reaching the tip and I+ blue flanks merging at the ascus tip (Fig. 3A,B). The “Caliciaceae” are part of the Buellia-clade, and their thin-walled, “prototunicate” ascus has been interpreted as a reduction of the Bacidia-type (Helms 2003). Iodine reactions of the ascus apex correspond very well with the ascus ultrastructure and are still the most convenient way to distinguish the two ascus types. In the light microscope the functionally arrested-bitunicate ascus is characterized by a rigid exoascus and an expanding endoascus. The exoascus ruptures during dehiscence and the tholus, which is part of the endoascus, expands, shooting out of the ascus and forming a short rostrum. In the Physciaceae only the endoascus and the mucilaginous surface of the ascus stain with iodine. The exoascus remains unstained and is a thin, uniform layer. Both the endo- and exoascus are functional rather than distinct structural layers of the ascus. Honegger 1978, 1987) examined the ultrastructure of some lecanoralean asci in detail. 37 The transmission electron microscope reveals that the structure of the ascus wall is more complex, and up to five layers can be distinguished during the ascus ontogeny (Fig. 3D-E; labeled a, b, c, d1 and d2 according to Bellemère 1994). In some asci the a- and blayer function as the exoascus, in other asci the a-, b- and part of the c-layer function as the exoascus (Bellemère 1994). The a- and b-layer were usually not well preserved in specimens of Buellia prepared for electron microscopy and it may be possible that the exoascus is mostly formed by the c-layer (Fig. 3E). The d-layer differentiates during ascus ontogeny to form the tholus. The outer I+ blue ascus flanks of the Bacidia-type in Buellia s.l. corresponds with the d1-layer and the central, unstained part is the d2-layer (compare Fig. 3A with 3E). The different layers are not easily distinguished in the TEM because they do not stain very differently with conventional stains like OsO4, uranyl acetate or lead citrate. Software programs like Adobe Photoshop allow subtle changes in contrast and help reveal the different structures of the layers. Only the outermost layers, the a and b layer are distinctly paler than the inner c and d layers. The outer part of the tholus, the d1-layer can be distinguished from the c-layer because its a fibrose to lamellate structure. In contrast, the innermost part, the d2 layer, is homogenous throughout. A very distinct ocular chamber is developed in young asci but disappears with maturity. Iodine staining corresponds very well to these structural observations, but it needs to be emphasized that the ascus is subject to a dynamic development, which considerably affects the structure of the tholus. The distinct differences in the tholus of the Lecanoraand Bacidia-type are best seen in asci with immature, unstained spores. At this early 38 stage, the spores are small and the tholus is not subject to much pressure. With maturity spores become larger, the ascus swells, and the tholus becomes flattened by increasing pressure. The tholus flanks and the central, non-amyloid cone thus often become deformed in mature asci. Asci are best examined from squash preparations of thin apothecial sections, but excessive pressure can result in considerable artifacts and great care must be taken to examine only intact asci. Fluorescent stains show great promise for revealing internal ascus structure without pressure artifacts, but experiments with various stains have so far failed to reveal different parts of the tholus. Some stains like calcoflour white react with the outer ascus wall and part of the ascospore walls, but do not differentiate separate layers within the tholus (Fig. 55C). ASCOSPORES All Physciaceae, including the calicioid taxa (Wedin & Grube 2002), have very distinctly layered and deeply pigmented ascospores (Fig. 4A-H). Spores of other lecanoralean lichens are usually less differentiated (Fig. 4I). Spore septa commonly develop soon during the spore ontogeny, but are not present in all taxa, especially some calicioid Physciaceae. Especially some calicioid Physciaceae also have non-septate ascospores. Spore Ultrastructure The spore ultrastructure in the Physciaceae is perhaps the most diagnostic character for an emended family concept which includes calicioid taxa (Wedin et al. 2000). Spore formation in ascomycetes is a unique process referred to as free-cell formation (Harper 39 1897). During this process ascoplasm becomes cleaved into vesicles by an enveloping membrane system (Czymmek & Klomparens 1992). These vesicles are initially open but soon enclose the new spore cells. Each vesicle is surrounded by two membranes (Beckett 1981). The outer membrane is referred to as the spore investing membrane. It segregates the epiplasm from an inter-membrane space. The inner membrane is called the spore plasma membrane, which separates the sporoplasm from the inter-membrane space. During spore wall ontogeny the inter-membrane space becomes converted into several distinct layers, which form the spore wall. Both the “investing” and the spore plasma membrane apparently contribute to this formation of the cell wall. The process is probably shared by all ascomycetes. Both membranes are not easily distinguished in Physciaceae spores prepared for the TEM. The outer membrane probably disintegrates as soon as the spore wall is fully matured. All inner membranes including the spore plasma membrane and the membranes of the cell organelles are rarely well preserved, because the thick spore wall is a considerable obstacle to rapid diffusion into the cell by chemical fixatives. Nordin (1997) pioneered the research on spore ultrastructure in the Physciaceae. He distinguished four different layers: a perispore, an intermediate layer, a proper spore wall and an endospore. In a revised concept Nordin & Mattsson (2001) confined the perispore to the “persistent part of the outermost spore wall”, distinguished from the mucilaginous sheath, which may sometimes be distinguished in the light microscope as a thin halo. In mature spores, most of these layers can usually be distinguished but not all of the layers are present in all species. 40 → FIGURE 4. Comparison of ascospore ultrastructure in the Physciaceae (calicioid and non-calicioid) and the Lecanoraceae [A-D. Thelomma mammosum (calicioid Physciaceae, Bungartz 3125, hb. Bungartz), E-H. Buellia navajoensis (non-calicioid Physciaceae, Nash 1577, ASU–holotype), I. Lecidella asema (Lecanoraceae, Nash 20954, ASU)].—A. Mazaedium of Thelomma: the asci disintegrate and form a mass of spores and paraphyses.—B. Close-up of the mazaedium: (s) spores, (p) paraphyses.—C. Ascospore of Thelomma.–D. Wall layers of a mature ascospore: (1) perispore, (2) intermediate layer, (3) proper wall, (4) endospore.—E. Immature ascospore of Buellia: (1) primary wall. (2) secondary wall.—F: Wall layers of an immature ascospore: (1) primary wall (perispore and mucilaginous sheath not distinctly differentiated); (2) secondary wall beginning to differentiate into the proper spore wall (p) and the endospore (e).—G. Mature ascospore of Buellia.—H. Wall layers of a mature ascospore: (s) mucilaginous sheath, (1) perispore, (2) intermediate layer, (3) proper wall, (4) endospore.—I. Mature ascospore of Lecidella: (1) primary wall, (2) secondary wall. 41 42 → FIGURE 5. Ultrastructure of paraphyses, hyphae and ascospores in the Physciaceae.— A. The epihymenium of Buellia sequax (Nash 32187, ASU): Several branching paraphyses with distinct pigment caps.—B. Paraphysal end cell with a distinctly layered, electron-dense pigment cap (Buellia sequax, Nash 32187, ASU).—C. Septal pore of an ascospore in Buellia dispersa (Nash 40593, ASU): the proper septum is formed by the proper wall (1) and the endospore (2); a canal (c) connects the two cell lumina but is blocked by a simple septal pore plug (p).—D. Ascospore of Buellia tergua (Nash 38412, ASU): a proper septum (s) with a septal pore (arrow) separates two distinct spore cells.— E. Somatic hypha of Buellia halonia (Nash 32718, ASU): (c) cell wall, (s) septum, (p) pore plug.—F. Septum of a somatic hypha (Buellia halonia, Nash 32718, ASU): the pore plug (p) is a simple, electron-dense structure, it appears to be delimited on the outside by a single membrane (arrow). 43 44 Differentiation of the cell wall begins with a differentiation into an inner secondary wall and an outer primary wall, both separated by an intermediate layer (Fig. 4E,F). At the early stages of the wall ontogeny this intermediate layer is barely visible, but it usually becomes distinct in mature spores (Fig. 4G). The outer primary wall differentiates into a mucilaginous sheath from the inner primary wall, which forms the perispore (Fig. 4F,H). Most species of Buellia s.l. form a distinct perispore. With maturity the perispore can become finely fissured or coarsely fractured, thus forming an ornamentation often visible in the light microscope. However, Nordin (2000) mentioned that in some species the perispore can be very thin or even absent. Buellia regineae is a species from the Sonoran Desert Region with a very thin perispore (Fig. 49F). In contrast, B. trachyspora has a considerably thickened and very strongly fractured perispore (Fig. 41F). The secondary cell wall matures into the proper spore wall. In the Physciaceae an endospore differentiates from this inner wall. The endospore development is quite dynamic throughout the development of the spore and responsible for the formation of distinct shapes of the cell lumina. In general, endospore thickenings correspond well to the wall and septal thickenings observed with the light microscope. However, some species show considerable endospore thickenings (e.g. B. badia), which have not been observed with the light microscope. Spores in the Physciaceae were frequently referred to as polarilocular if a distinct septum thickening is present and a connection of the two spore cells is visible (Sheard 1967; Øvstedal & Lewis Smith 2001; Kalb 2002; Nimis & Tretiach 2002). It might be suggested that these spores resemble the polarilocular ascospores of the Teloschistaceae, 45 and indeed Poelt (1973) included the Teloschistaceae and the Physciaceae within the suborder Buelliineae. This assessment is not supported by the ultrastructure of the spores. In all Physciaceae spore septa form from both the endospore and the proper spore wall. These septa therefore separate two distinct cells. Like the somatic cells of ascomycete hyphae, the cell lumina of the spore cells remain connected by a septal pore (Fig. 5C-F). In Buellia this pore is soon blocked by an electron dense, undifferentiated plug (Fig. 5C,D). Swelling of the septum is generally confined to the endospore and a distinct pore canal develops in spores with a thickened septum (Fig. 5C). Spore septa of the Teloschistaceae form only from an endospore swelling, and the proper wall does not become part of the septum (Bellemère & Letrouit-Gallinou 1982). The endospore swelling of these spores initially does not separate two cells, but divides the cell lumen of one single cell into two locules. The two locules of the cell remain connected by an isthmus during much of the ontogeny. A new cell membrane with a pore plug is only inserted at the very last stage of the septum formation. Spores of the Teloschistaceae with an endospore swelling are therefore correctly referred to as polarilocular (i.e. one-celled with several locules), whereas the Physciaceae clearly have two-celled, non-polarilocular spores. Spores of the Physciaceae are sometimes referred to as “mischoblastiomorphic” (Kalb 2002) but the term may be confused with the Mischoblastia-type ascospores of Rinodina. The Dictionary of the Fungi (Hawksworth et al. 1995, p. 281; Kirk et al. 2001, p. 325) repeatedly defined “mischoblastiomorph” as “… similar to the polarilocular type but either without a septum or only with an incomplete septum.”. The ultrastructure of the 46 spore septum in the Physciaceae does not conform with this definition! The presence of a septal pore does not indicate that septum development is “incomplete”. Purvis (1992) suggested that Diplotomma could be distinguished from Buellia by the formation of distosepta. Nordin (1997) examined the ultrastructure of ascospores in Buellia s.l. and compared species with pluriseptate and one-septate spores. He demonstrated that even in species with pluriseptate spores most septa were formed both by the proper wall and the endospore. Luttrel (1963) first introduced the term “distoseptum” to describe conidial septation of non-lichenized fungi. Hawksworth et al. (1995) later also used the term referring to endospore septa in Caloplaca. Nordin (1997), however, argued that the ultrastructure of “distosepta” formed within conidia was not necessarily analogous to the ultrastructure of septum formation in ascospores. The studies presented here agree well with Nordin's (1997) assessment (Fig. 5C,D). Spore Ontogeny and Spore Types Ascospores of many Physciaceae develop conspicuous wall thickenings during some stages of their spore ontogeny. These septal, apical and lateral thickenings have been used to define many different spore types and they are extensively used, especially in the taxonomy of Rinodina s.l. (Mayrhofer & Poelt 1979; Mayrhofer 1982, 1984b; Giralt & Mayrhofer 1994b; Giralt 2001). Classical concepts of Buellia have long ignored wall or septum thickenings because they are usually less prominent. Most species of Buellia thus have Beltraminea (= Buellia)-type ascospores. Scheidegger (1993), however, emphasized septum thickenings in some species of Buellia and related genera. He assigned these spores to the Physconia-type, a type also present in Rinodina. Lateral wall thickenings of 47 the Callispora-type are characteristic for “Hafellia” (Scheidegger 1993). Some species of “Amandinea” with pronounced septal thickenings might be referred to the Orculariatype (Mayrhofer et al. 1999; Bungartz et al. 2004b, Publication 5). Unfortunately, these type-concepts are quite static. The spore types are essentially based on a hypothetical “representative” stage during the ascospore ontogeny. This stage is usually referred to as the mature spore and is deemed to be characteristic for a particular type. The types may be convenient for identification of taxa, but do not necessarily agree with the phylogeny of the species. Thus types like the Physconia-type must be considered homoplastic (Helms 2003) because the characteristic septal thickening obviously occurs independently, both in the Buellia-clade (clade B sensu Helms 2003) and in the Physcia-clade (Rinodina-clade, clade A sensu Helms 2003). To accommodate the dynamics of the spore ontogeny Giralt & Mayrhofer (1994a; 1995) distinguished two basic ontogenies based on their septum formation. Ascospores display early septum formation (A-ontogeny), if their septa form before apical thickenings are visible. During late septum formation (B-ontogeny) apical thickenings can be observed prior to septum insertion. However, spores in Buellia s.l. often lack apical thickenings. The concept must therefore be modified if it is applied to Buellia. Bungartz et al. (2004b, Publication 5) suggest that spore wall pigmentation may correlate with the ontogeny of the septum formation. Thus, spore septa either develop before the spore walls become pigmented (A-ontogeny) or later, when the wall is at least faintly pigmented (B-ontogeny). Even this concept does not appropriately describe the complex 48 ontogeny of all ascospores and in some species spores cannot be clearly assigned to either A- or B-ontogeny. Towards a more dynamic concept of spore ontogeny in the Physciaceae At least four stages of a dynamic spore ontogeny can be disinguished in the Physciaceae (Bungartz et al. 2004b, Publication 5): (1) Immature ascospores are generally unpigmented and no ornamentation can be observed, they often have no septum (A-ontogeny) or a thin septum becomes visible at the end of this stage (B-ontogeny). Apical wall thickenings can sometimes be observed at this stage. (2) Premature ascospores become increasingly pigmented from faint green to dark olive, and in some species ornamentation becomes visible. The septum is usually present at this stage and characteristic wall thickenings often begin to develop. (3) Mature ascospore usually have a brown to deep brown pigmentation and wall ornamentation is usually distinct at this stage, although some spores lack spore ornamentation during their entire ontogeny. Characteristic wall thickenings are most characteristic at this stage, although septum thickenings are often more pronounced in the previous, premature spores. (4) Overmature spores are always deep brown and if any ornamentation is present it is most visible at this stage. In Buellia s.l. wall thickenings again become reduced. Many spores retain some of the thickening, but the characteristic shapes of the cell lumina are much less distinctive. If spores are not ejected from the asci they begin to deteriorate, and old overmature spores usually have damaged and disintegrating cell walls. Disintegrating 49 spore walls often have a false ornamentation, artifacts caused by the deterioration of the spore wall. The dynamics thus described have considerable implications for the distinction of spore types. Spore ornamentation, septal and wall thickenings, and pigmentation all develop independently. In some species, ornamentation can only be observed at a late stage, in others it is formed early during the ontogeny. Characteristic wall or septal thickenings are rarely persistent, and only a few species retain a ± diagnostic shape of their cell lumina for considerable periods of time. Only spores of these species can easily be assigned to a particular type, but in the majority of species wall and septal thickenings are much more dynamic. A diagnostic and distinctly thickened septum is thus often only observed during brief periods of the ontogeny. This is particularly true for spores of Buellia s.l. assigned to the Physconia-type by Scheidegger (1993), and several of the species studied here. Buellia dispersa is a good example with a very dynamic development. Septum thickenings are typically only observed during a brief period of the spore ontogeny. This is also the case in B. nashii, B. paniformis, B. navajoensis and several other species examined from the Sonoran Desert Region. In Buellia regineae lateral wall layers can only be observed very briefly and most spores are thus evenly thin-walled, even though the species is closely related to “Hafellia”. Only a few species from the Sonoran Region have a ± persistent septum thickening, e.g., B. halonia, B. lacteoidea and B. subdisciformis. In B. prospersa the septum is 50 considerably thickened during most of the spore ontogeny, but even in this species some specimens have spores with considerably less pronounced septal thickenings. Conclusions The Physciaceae sensu Wedin & Grube (2002) can thus be characterized by ascospores with distinctly pigmented, layered cell walls with a dynamic wall and septum development. They are rarely non-septate but more frequently form proper septa. In a few species additional endospore septa can be observed, but these are not confined to a particular group and even occur in some species with predominantly one-septate spores (e.g., B. argillicola, B. concinna, B. trachyspora). Septum formation, wall differentiation and pigmentation all are subject to change and are not easily categorized. Especially the differentiation of the different wall layers is a very dynamic process. In a few species the perispore remains thin or cannot be observed at all, but many species have a strongly developed perispore, often distinctly ornamented at an early or late stage of the ontogeny. The endospore is as variable as the perispore and responsible for apical, lateral and septal thickenings. These thickenings are not static but change quite dynamically during spore ontogeny. Spore types in the Physciaceae need to be revised to accommodate all stages of ascospore development. Pycnidia All conidiomata of Buellia s.l. are pycnidia (Fig. 6A,B,E). Vobis (1980) distinguished five different types of pycnidia primarily based on their ontogenies, and to some extend also on the types of conidiophores. He examined several species of the Physciaceae and 51 assigned their pycnidia to the Umbilicaria-, Lobaria-, or the Roccella-type. Interestingly, the only representative of the “Caliciaceae” examined by Vobis (1980) was also characterized by Umbilicaria-type pycnidia. The only species of Buellia examined by Vobis (1980) is Buellia stillingiana J. Stein., which he did not assign to a particular type, even though his illustrations suggest that the pycnidium is very similar to the Umbilicaria-type. Marbach (2000) argued that Buellia triphragmoides was the only species of Buellia s.l. characterized by Umbilicaria-type pycnidia. He therefore included this species within the monotypic genus Mattickiolichen. Marbach (2000) suggested that all other species of Buellia s.l. with bacilliform conidia were characterized by Lecanactis-type pycnidia. According to his concept, pycnidia of the Lecanactis-type would be distinguished by sparsely branched, parallel conidiophores, whereas the Umbilicaria-type pycnidia would be described by branched conidiophores growing in all directions. These descriptions do not conform with the types described by Vobis (1980), who distinguished pycnidia not by the orientation of their conidiophores, but by the ontogeny of the entire pycnidium. All pycnidia examined here do not develop into an open, cupulate structure with unbranched type II conidiophores. Thus, they do not belong to the Lecanactis-type sensu Vobis (1980). Instead all pycnidia in Buellia s.l. are flask-shaped, opening by a distinctly pigmented pore, and they are lined with moderately to densely branched conidiophores (Fig. 6A,B,E). The orientation of the conidiophores is primarily a result of the branching pattern. Thus, moderately branched conidiophores are predominantly parallel, whereas conidiophores, which are more abundantly branched, are not as clearly oriented. Even 52 within a single pycnidium, some conidiophores are often less branched and more parallel than others. Scheidegger (1993) first recognized that some species of Buellia s.l. (and Rinodina s.l.) were characterized by long, filiform conidia (Fig. 6F). He suggested that these filiform conidia were borne on conidiophores different from Buellia s.str., and are characterized by short, bacilliform conidia (Fig. 6C,D). Scheidegger (1993) considered these differences sufficient to provide a valid description of Amandinea, a genus previously introduced by Choisy (1950). Several species were subsequently transferred into this new genus (Sheard & May 1997; Marbach 2000) and the transfer was mostly justified by occurrence of filiform conidia. Marbach (2000), however, also included several species without knowing whether these had filiform conidia. He argued that Sheard & May (1997) had provided an emended genus concept, which justified this decision. It can be demonstrated, however, that at least the transfer of B. turgescens is not justified, because that species is synonymous with Buellia badia, which is characterized by short, bacilliform conidia Bungartz & Nash (2004b, Publication 2). In the Sonoran Desert Region, many inconspicuous saxicolous species were often assigned to Buellia (= Amandinea) punctata even though most of them have bacilliform conidia (Bungartz et al. 2004b, Publication 5). Measurements of conidial length considerably overlap (Fig. 31). It is therefore problematic to distinguish conidia only by their length. If the shape is also taken into account, bacilliform, fusiform and filiform conidia might be distinguished. In general 53 conidiophores are more distinctly branched if they bear short, bacilliform conidia. They are less frequently branched in species with fusiform conidia and almost entirely unbranched in species with filiform conidia. None of these differences can be considered very significant. Long, filiform conidia are necessarily formed on short, unbranched conidiophores, otherwise they would become entangled within the branches and could not be released through the ostiole. Short, bacilliform conidia can be produced in larger quantities, if these are not only formed from terminal cells but also from side branches or bayonet-like projections. Pycnidia with filiform conidia are therefore not qualitatively different from ones with bacilliform conidia (Fig. 6). They are all flask-shaped, opening with a distinct ostiole, densely lined with relatively short, sparsely branched conidiophores terminally forming fusiform or filiform conidia, or moderately branched, forming conidia from terminal cells, side branches and bayonet-like projections. With increasing length of the conidia, conidiophores become less elaborately branched, the bayonet-like projections are less frequent and the length of the conidiophores is reduced. Pycnidia in Buellia s.l. therefore form a natural progression from the Umbilicaria-type to the Roccella-type. In its most recent circumscription several very different species are unified within the genus Amandinea because of their filiform conidia. Some of theses species have a thalline exciple, others lack a thalline exciple; some species have hyaline, others a pigmented hypothecium;some species have spores with septal thickenings, others have spores without septal thickenings; some species contain lichen substances, others entirely lack secondary metabolites. It is not conceivable that Scheidegger (1993) had anticipated 54 that this new genus soon would be as heterogeneous as Buellia s.l. and the current concept of Amandinea is not very convincing. → FIGURE 6. Pycnidia, conidiophores and conidia of Buellia s.l.—A. Umbilicaria-type pycnidium of B. tergua (Nash 38368, ASU–holotype).—B. Umbilicaria-type pycnidium of B. christophii (Nash 33979, ASU–holotype).—C. Conidiophores of B. ryanii with bacilliform conidia (Nash 32448b, ASU).—D. Conidiophores of B. tergua with bacilliform conidia (Nash 38368, ASU–holotype).—E. Roccella-type pycnidium of B. pullata (Nash 41227, ASU).—F. Conidiophores of B. prospersa with filiform conidia (Wetmore 71896, COLO). 55 56 CHEMISTRY As part of this research almost all specimens were routinely examined with TLC and some then selected for a more detailed analysis with HPLC (APPENDIX I). Results of these analyses reveal a diverse chemistry of secondary metabolites (Table 1). Like all other characters, secondary chemistry provides very valuable information, but usually does not suffice to distinguish species from one another. Most chemotypes correspond clearly with species recognized by other classical characters like thallus morphology, apothecial or pycnidial anatomy or geographic distribution. For example, B. spuria can reliably be distinguished from B. stellulata by its secondary chemistry. The two species are also distinguished by different iodine reactions and differences in distribution. Buellia spuria contains norstictic acid, has a I+ blue medulla, and is widely distributed. The medulla of B. stellulata does not react with iodine, it contains 2'-O-methylperlatolic instead of norstictic acid, and has an oceanic distribution. Another example, where the exact same substances are diagnostic, is the distinction of B. dispersa and B. nashii. Buellia dispersa is characterized by 2'-O-methylperlatolic acid and lacks the aeruginose exciple pigment cinereorufa-green. The species is very widely distributed throughout the Sonoran Region. In contrast, B. nashii is less widely distributed, is always characterized by an aeruginose outer exciple, and contains norstictic acid. Other species have traditionally been segregated as distinct species because of their chemistry, but more detailed studies show that their chemistry sometimes overlaps. Thus, 57 Scheidegger (1993) distinguished B. tyrolensis (with norstictic acid) from B. fusca (with 2'-O-methylperlatolic acid). Analysis of a larger amount of specimens, however, revealed, that specimens with both substances exist and no other morphological, anatomical or distributional characters can be used to distinguish the two taxa (Bungartz 2004, Publication 8). Consequently, B. fusca must be treated as a synonym of B. tyrolensis, even though the exact same secondary metabolites have proven very useful in distinguishing B. spuria from B. stellulata or B. dispersa from B. nashii, respectively. Several species with a similar secondary chemistry are not necessarily closely related. For example, species with xanthones display a considerable variety of different anatomical and morphological characters, and it is unlikely that they constitute a natural evolutionary group (Bungartz et al. 2004c, Publication 7). Most species are characterized by the presence of a few typical metabolites but some species show a rather complex chemistry. They contain a variety of metabolites, which are not always present in every specimen of a species. Many metabolites occur as chemosyndrome, i.e. they are often metabolically related and thus occur in characteristic combination. Usually at least one substance of a chemosyndrome is regularly and easily detected as the major substance, while other substances of the same syndrome are missing or remain undetected due to low concentrations. However, even common and characteristic, major substances are sometimes lacking or only present in low concentrations. The absence of characteristic spot test reactions is therefore not reliable to identify all specimens even though the majority of specimens display a distinct reaction. 58 Some secondary metabolites can be confined to different parts like the thallus cortex, the medulla or the apothecia. Atranorin, xanthones or usnic acids are characteristic for the cortex and probably provide a protective UV-screen especially for the photobiont layer (Nash 1996). Norstictic acid is often confined to the medulla but has also been found in the apothecia. All these substances are acetone soluble and thus detected by TLC or HPLC. Acetone insoluble apothecial pigments have been discussed in detail in the context of exciple pigmentation (for the terminology see Meyer & Printzen 2000). These pigments can be distinguished by a series of color shifts with reagents like HNO3, KOH and HCl but the distinction is not entirely reliable if acetonesoluble secondary metabolites like atranorin or norstictic acid are also present. Table 1 gives an overview of the secondary metabolites found in species with one septate ascospores in the Greater Sonoran Desert Region. In general these substances can be arranged in several substance classes: Orcinol depsides A few species of Buellia s.l. contain orcinol depsides, of which 2'-O-methylperlatolic acid, sometimes accompanied by confluentic acid, is the most common. Other orcinol depsides have only been found in a few or even a single species: Gangaleoidin and the closely related dechlorogangaleoidin and norgangaleoidin have only been found in B. regineae. Brialmontin 1 and chlorolecideoidin also occur only in this species. Nordin (2000) reported brialmontin 1 & 2 from B. muriformis, a corticolous and lignicolous species with pluriseptate spores. Marbach (2000) reports brialmontin 1 & 2 from some corticolous Hafellia-species. Buellia regineae also contains several orcinol depsidones 59 commonly associated with the genus Hafellia. Gyrophoric and lecanoric acid are two closely related orcinol depsides, which are fairly common in lichens. They occur only in a few species examined here: Buellia eganii, B. lacteoidea, B. subdisciformis and B. uberior. Buellia tesserata is the only species with 3-chlorodivaricatic acid, a chemistry shared with the closely related Dimelaena radiata. Orcinol depsidones Diploicin, fulgidin and related orcinol depsidones are restricted to three species: Buellia lepidastroidea, B. paniformis and B. regineae. These orcinol depsidones and closely related substances are characteristic for several species with oil inspersion in the hymenium, as those in “Hafellia” or the Buellia excelsa-group. Diploicin was first discovered in Diploicia, which Molina et al. (2002) suggested synonymizing with Diplotomma based on molecular evidence. Nordin's (2000) cladistic analysis did not agree with this concept. Diploicia instead grouped with other species also containing diploicin. β-Orcinol depsides The only β-orcinol depsides currently known from Buellia s.l. are atranorin and chloroatranorin. With HPLC several artifacts caused by the hydrolysis of atranorin were also detected: haematommic acid and methyl-β-orsellinate. Both atranorin and chloroatranorin are common in Buellia s.l. and also occur in many other not necessarily related lichen species. 60 β-Orcinol depsidones Both the norstictic and the stictic acid chemosyndromes are among the most common secondary lichen metabolites. Norstictic acid is typically associated with connorstictic acid. Stictic acid is part of a more complex group of secondary metabolites including constictic, hypostictic, cryptostictic and methylstictic acid. Only a few species of Buellia s.l. lack β-orcinol depsidones. Xanthones Xanthones are cortical pigments common to several species (Scheidegger & Ruef 1988; Bungartz et al. 2004c, Publication 7). They also occur as pruina on the surface of the thallus or apothecia. They typically cause a pale yellow to yellow green thallus color and can often be detected by a yellow to orange fluorescence in long-wave UV-light. This fluorescence may not be very bright in specimens with low concentrations. Many, but not all xanthones react C+ orange. The following species contain xanthones: Buellia concinna, B. halonia, B. mamillana, B. navajoensis, B. prospersa, B. subaethalea and B. trachyspora. Xanthones are also known from other genera of the Physciaceae: Dimelaena, Diploicia, Pyxine and Rinodina as well as many other lichens. Usnic acid related substances Although a very common lichen substance, usnic acid rarely occurs in the Physciaceae. It is characteristic for some species of Dimelaena and has been reported from a few species with pluriseptate spores (Nordin 2000). The closely related isousnic acid often occurs with usnic acid. Another closely related substance is placodiolic acid. 61 Unlike usnic or isousnic acid, placodiolic acid is generally a rare lichen substance. Scheidegger (1993) reported placodiolic acid from Buellia (“Hafellia”) leptoclinoides, which does not occur in the Sonoran Desert Region. The closely related B. regineae instead contains the orcinol depsidone diploicin. The saxicolous B. capitis-regum also contains placodiolic acid. The species may thus be closely related to “Hafellia”, although it has pluriseptate spores (see the discussion on phylogeny). Anthraquinones The ergochromes eumitrin x & y have been detected by HPLC in specimens of B. halonia. The have also been found in one specimen tentatively identified as B. subdisciformis. A specimen of B. christophii contains parietin but the substance was not found in any other specimen examined and is most likely a contaminant. Anthraquinones are generally not common in the Physciaceae, although B. capitisregum contains the deep orange, K+ purple anthraquinone cinnanomeic acid in its medulla. This species with pluriseptate spores is common along the coast of the Sonoran Desert Region. Marbach (2000) resurrected the genus Gassicurtia because of the presence of orange-red anthraquinones in medulla and exciple. Grube et al. (2004) reported the reddish anthraquinones eumitrin U and secalonic acid derivates from terricolous specimens. Other substances The dibenzofurane virensic acid was detected by HPLC in B. regineae but has not been reported in any other species. Scheidegger (1993) reported barbatic acid from B. 62 tesserata, but Rico et al. (2003) re-examined the type specimen and concluded that the substance had been misidentified. Instead of barbatic acid they reported 3chlorodivaricatic acid and therefore synonymized B. fimbriata with B. tesserata. 63 TABLE 1. Secondary metabolites in Buellia s.l. (saxicolous species with one-septate ascospores in the Greater Sonoran Desert Region). B. aethalea B. amabilis B. argillicola B. badia B. christophii B. concinna B. dispersa B. eganii B. halonia B. lacteoidea B. lepidastroidea B. mamillana B. nashii B. navajoensis B. paniformis B. prospersa B. pullata B. reginea B. ryanii B. sequax B. sheardii B. spuria B. stellulata B. subaethalea B. subalbula B. subdisciformis B. tergua B. tesserata B. trachyspora B. tyrolensis B. uberior B. vilis X X X X Orcinol- (x) (x) (x) X X X X Orcinol- X X X (x) depsidones (x) X X X X X depsides no substances found (x) no substances found X X (x) no substances found X X no substances found rarely with substances X rarely without substances no substances found X X X (X) no substances found X X X X X (x) (x) X X X Xanthones X X lichexanthones X X X X X X (x) X (x) X (x) X X (x) (x) X (x) X X (x) (x) (x) (x) (x) (x) (x) (x) (x) (x) (x) ȕ-Orcinol- (x) (x) (x) (x) (x) (x) (x) (x) X (x) X X X X X X X X X X (x) X X X (x) (x) X (x) X X (x) X X (x) X X X (x) X X (x) (x) (x) (x) (X)(x) X norlichexanthones X X X X X X X X others X X X X X X X X X ? X ? ? ? Anthraquinones ? ergochromes X X X X ? X 2’-O -methylperlatolic 3,5-dichloro-2'-O -methylanziaic acid 3,5-dichloro-2'-O -methylnorimbricaric 5-O -methylhiascic acid brialmontin 1 chlorolecidioidin confluentic divaricatic 3-chlorodivaricatic nordivaricatic gangaleoidin dechlorogangaleoidin norgangaloidin gyrophoric lecanoric orsellinic diploicin dechlorodiploicin 3-dechlorodiploicin 3-demethylscensidin fulgidin isofulgidin depsides atranorin chloroatranorin depsidones norstictic connorstictic stictic constictic hypostictic cryptostictic methylstictic 4-chlorolichexanthone 2,4-dichloronorlichexanthone 2,5-dichlorolichexanthone 4,5-dichlorolichexanthone 2,4,5-trichlorolichexanthone 2,4-dichloronorlichexanthone 2,5-dichloronorlichexanthone 2,7-dichloronorlichexanthone 4,5-dichloronorlichexanthone 5,7-dichloronorlichexanthone 2,7-dichloro-6-O -methylnorlichexanthone 4,5-dichloro-3-O -methylnorlichexanthone arthothelin isoarthothelin 6-O -methylarthothelin asemone thiophanic thiophaninic thuringione eumitrin x eumitrin x2 eumitrin y parietin secalonic Dibenzofurans virensic acid 64 DISTRIBUTION AND ECOLOGY Distribution The Greater Sonoran Desert Region as outlined by Nash et al. (2002) covers a large area from central Sinaloa to northern Arizona and southern California. It is centered around the Sonoran Desert as circumscribed by Shreve & Wiggins (1964). It is characterized by considerable differences in topography, climate and geology (Phillips & Wentworth Comus 2000). The elevation ranges from sea level along the coast of Southern California and the Baja California peninsula, through the Lower Colorado Basin (ca. 300-600 m) up to the Colorado Plateau (ca. 2000 m), peaked by a few isolated volcanoes like the San Francisco Peaks (3600 m). The climate along the northern coast is Mediterranean with mild, wet winters and warm, dry summers but farther south summer precipitation predominates. Towards the central part of Arizona rainfall becomes scarce and occurs in the winter, when cyclonic storms migrating from the Pacific ocean move across, and summer thunderstorms are associated with the influx of moisture from the south (Sellers et al. 1985; Ingram 2000). The northeastern Colorado Plateau has a temperate, continental climate with cold and dry winters. The Sierra Madre Occidental of Chihuahua, Sonora and Sinaloa as well as Southern Baja California are subject to more subtropical influences. with precipitation becoming more restricted to summer months. The geology of the region is dominated by siliceous rocks with little or no carbonates (Nations & Stump 1983). Volcanic, igneous rocks like basalt, rhyolites and andesites are among the most common substrates of this geologic region transversed by an active geologic fault line (Chronic 1983). Sedimentary rocks, especially sandstones, are also 65 common, especially on the Colorado Plateau, which was lifted from the bottom of an ancient sea as a large block of sediments. Among these sediments limestones are rare and gypsum is very scarce. As a result of evaporation, some secondary carbonates are present in the Basin and Range areas of the Lower Colorado Desert (McPhee 1990). At the heart of the region lies the Sonoran Desert with its floristic subdivisions (Shreve & Wiggins 1964). The region lies in the middle of the North American Desert Regions. It merges with the temperate Mojave Desert in the northwest and the subtropical Chihuahua Desert in the east. The Sonoran Desert is probably the most diverse region of these three deserts as a result of this overlap (Brown 1994). The paleontology of the Sonoran Desert suggests that the current desert vegetation is relatively recent, probably less than eight million years of age (Devender 2000). Species currently present in this area have thus had little time to adapt. The present vegetation is composed largely of species, which must have originated either in a northern, temperate or a southern subtropical climate but were able to tolerate a dry climate, when a general drying trend began fifteen to eight million years ago. Species diversity in the genus Buellia reflects the diversity of the region. Many taxa are considered strictly coastal, maritime species: Buellia christophii, B. halonia, B. lepidastroidea, B. paniformis, B. pullata, B. regineae, B. subalbula, and B. tesserata. Buellia prospersa also has a distinct maritime tendency but may also be distributed further inland. Buellia tergua and B. ryanii are probably restricted to the shore but only a few specimens of these species are currently known. 66 → FIGURE 7. Distribution of Buellia aethalea, B. argillicola, B. badia, B. christophii, B. concinna and B. dispersa in the Greater Sonoran Desert Region. 67 68 → FIGURE 8. Distribution of Buellia eganii, B. halonia, B. lacteoidea, B. lepidastroidea, B. mamillana and B. nashii in the Greater Sonoran Desert Region. 69 70 → FIGURE 9. Distribution of Buellia navajoensis, B. paniformis, B. prospersa, B. pullata, B. regineae and B. ryanii in the Greater Sonoran Desert Region. 71 72 → FIGURE 10. Distribution of Buellia sequax, B. sheardii, B. spuria, B. stellulata, B. subaethalea and B. subalbula in the Greater Sonoran Desert Region. 73 74 → FIGURE 11. Distribution of Buellia subdisciformis, B. tergua, B. tesserata, B. trachyspora, B. tyrolensis and B. uberior in the Greater Sonoran Desert Region. 75 76 → FIGURE 12. Distribution of Buellia vilis in the Greater Sonoran Desert Region. 77 78 Some species are considered oceanic, i.e. they grow close to the coast, but are not necessarily restricted to the shore. These species include B. stellulata and B. subdisciformis, and possibly B. trachyspora and B. prospersa. No species of Buellia s.l. is particularly well adapted to the arid central desert of the Lower Colorado Basin, and B. dispersa is the only species which is commonly found at low elevations. It is a very widely distributed, variable taxon, and it is possible that some of the morphotypes of B. dispersa are better adapted to the extreme conditions of the desert. The largest group of species may best be classified as montane: B. concinna, B. nashii, B. mamillana, B. lacteoidea, B. navajoensis, B. spuria, B. subaethalea, B. tyrolensis, and B. uberior are typically encountered at moderate to high elevations. In the Sonoran Region, Buellia vilis and B. aethalea appear to be restricted to higher elevations, and B. eganii has only been found at the highest, subalpine to alpine elevations. Some species, including Buellia badia, B. nashii and B. sequax, are not necessarily montane but they occur both along the coast and in the montane areas and may not be able to tolerate the lower desert extremes. Three floristic elements overlap in the Sonoran Region: temperate, Mediterranean (Madrean) and subtropical/tropical elements. Species like B. aethalea, B. concinna, B. stellulata, and B. vilis are of northern, temperate origin. These species are distributed throughout the Northern Hemisphere and are more common in colder climates than in the Sonoran Desert. Another group of species may be called Mediterranean. These species occur in Baja California and southern California and are also known from Mediterranean 79 Europe: B. dispersa, B. sequax, B. subalbula, B. tesserata, and B. tyrolensis all belong to this group. Buellia halonia, B. nashii and B. lacteoidea are also part of this group but are not known from Europe. Finally, some species clearly have a southern, subtropical to tropical origin: Buellia mamillana and B. subaethalea are subtropical species. Buellia sheardii is probably also subtropical but the distribution is currently not well known. Buellia argillicola has been described from central Mexico and very little is currently known about its distribution. Buellia trachyspora is a tropical species, first described from the West Indies. It barely extends into the southernmost part of the region. Ecology All species examined in this dissertation grow on rock. Buellia punctata, a species reported to grow on rock as well as bark (Scheidegger 1993), has not been confirmed from the Greater Sonoran Desert Region. A careful examination of all saxicolous specimens demonstrated that these specimens were generally misidentified as B. punctata and do not belong to this taxon (Bungartz et al. 2004b, Publication 5). Buellia badia is thus the only species also known from substrates other than rock, such as decorticated wood and even bark. Juvenile thalli of this species are facultative parasites on a wide range of host lichens, but with age establish independent thalli (Bungartz & Nash 2004b, Publication 2). In contrast, B. uberior is currently known only as an obligate parasite, forming gray insular patches in crustose lichens of the genus Schaereria. However, specimens from Novaya Zemlya described by Lynge (1928) as B. malmei are very similar but grow independently. From Europe B. concinna is known to 80 occur sometimes as a facultative parasite, but from the Sonoran Region only independent thalli are currently known. Most species examined here clearly prefer siliceous rock with no or only minor traces of carbonates. Only few species, like B. dispersa or B. nashii, can regularly be found on siliceous rocks with or without carbonates. Buellia amabilis, B. argillicola, B. navajoensis and B. subalbula prefer limestone substrates, or at least grow on carbonate rich sandstones. Saxicolous species with pluriseptate spores may be found more commonly on limestone than species with one-septate spores. Nordin (2000) suggested that the presence of large amounts of calcium oxalates constitutes an important taxonomic character for the Buellia alboatra-group (= Diplotomma s.str.). calcium oxalates are certainly not exclusive to this group but their presence in large amounts may suggest an adaptation to the presence of Ca carbonates in the substrate. Thalli of Buellia s.l. often contain large amounts of calcium oxalates if Ca carbonates are present within the substrate. However, some thalli contain calcium oxalates even if the substrate does not contain Ca carbonate. For example, thalli of B. dispersa and B. nashii usually contain at least small quantities of calcium oxalates, even when growing on rock reacting negative with 10 % HCl. As outlined in the previous chapter on distribution, most species from the Sonoran Region cannot be considered particularly well adapted to the desert climate. Many montane species grow in shaded canyons or narrow valleys often along ephemeral or permanent streams and small rivers. Even B. dispersa, which is found throughout the 81 entire region, does not often grow in exposed habitats. With increase in altitude, more rainfall is usually available and in the upper montane or subalpine to alpine habitats, species like B. eganii or B. aethalea can even be found in moderately exposed habitats. At lower elevations of the Sonoran Region, Buellia species are typically found on sheltered and shaded north sides of boulders, outcrops and cliffs. These habitats are usually more richly inhabited if they are located in proximity to a desert wash or stream, but they rarely occur close enough to be subject to inundation. It is not unusual for most desert lichens to prefer these sheltered habitats throughout the arid North American Southwest. Only a few soil crusts and small cyanolichens, especially the Lichinaceae, are adapted to withstand more extreme, arid conditions. Green algal lichens are capable of surviving without liquid water, but thus rely strongly on fog and dew (Nash et al.1982; Lange et al. 1990; Nash 1996). As a consequence species diversity of the genus Buellia is highest in the coastal fog deserts of Southern California and the Baja California peninsula. Species growing in this environment generally show exuberant growth. If they are growing in close proximity to the sea shore, thalli become subject to salt spray. These thalli are often considerably thicker than less exposed thalli. Sometimes extreme morphotypes of B. lepidastroidea develop an almost subfruticose appearance with “stalked” apothecia. The thallus surface of Buellia regineae becomes strongly “gnarled” and the typical, bullate thalli of B. paniformis (forming “bread-loaves”) may also reflect a morphological adaptation to the seashore habitat. 82 PHYLOGENY Results of the Analysis The cladistic analyses using morphological characters (table 2) consistently converged on a large number of equally most parsimonious trees of 385 steps. Retention, consistency and homoplasy indices of these trees were generally low. For example, tree no. 1 has a consistency index of 0.16, a homoplasy index of 0.84 and a retention index of 0.58. Only three nodes of the strict consensus tree (MAXTREES = 1000) were supported by non-parametric bootstrapping values greater than 50%, with Rinodina bischoffii and R. confragosa designated as the outgroup. Dimelaena radiata and D. oreina formed one group with high bootstrap support (86%). Buellia oidalea and B. oidaliella formed a second group with high bootsrap support (85%), and a third group of B. himalayensis and B. megaspora was weakly supported (51%). These results are comparable with the generally low support found by Nordin (2000) for a subset of the species analyzed here. The only groups supported in his analysis were the outgroup (R. bischoffii and R. confragosa), a group formed by B. oidalea and B. oidaliella and a group formed by B. alboatrior and B. submuriformis. Dimelaena radiata and Dimelaena oreina were not included in his data set. The data set examined here contained almost twice the number of taxa (n = 80) compared to Nordin’s (2000) original data set (n = 47). Nevertheless the strict consensus trees of both analyses are very similar. Although not supported by bootstrap values similar groups can be identified in both trees. 83 TABLE 2. Data matrix used for the cladistic analysis – = inapplicable, ? = unavailable character state Taxa Character numbers and states 1 11111 11112 22222 22223 33333 33334 44444 12345 67890 12345 67890 12345 67890 12345 67890 12345 Buellia aeruginosa Buellia aethalea Buellia alboatra Buellia alboatrior Buellia amabilis Buellia argillicola Buellia badia Buellia capitis–regum Buellia cedricola Buellia christophii Buellia concinna Buellia coniops Buellia disciformis Buellia dispersa Buellia eganii Buellia erubescens Buellia geophila Buellia graminicola Buellia granulosa Buellia griseovirens Buellia halonia Buellia himalayensis Buellia imshaugiana Buellia insignis Buellia jugorum Buellia lacteoidea Buellia lauricassiae Buellia lauricassioides Buellia lepidastroidea Buellia mamillana Buellia megaspora Buellia mexicana Buellia morsina Buellia muriformis Buellia nashii Buellia navajoensis Buellia oidalea Buellia oidaliella Buellia pallidomarginata Buellia paniformis 11100 11000 31010 01?00 11000 11000 30011 10000 01000 10000 10010 11200 00?00 10001 11000 01?00 20000 20?00 11000 01000 11000 00?00 01100 20?00 01101 11000 00000 01?00 11001 11000 00?00 11000 01?00 01000 10001 10000 01001 01001 01?00 10001 (cont.) 00000 11000 01011 00001 00011 01011 10001 10002 00001 00000 11001 10001 10001 10011 10000 10002 10011 10102 1?001 10102 10000 00002 1?001 00001 10000 11001 00002 10001 10011 01001 00001 10000 00001 00002 10011 11012 10002 10012 00001 10001 10111 300–0 10100 000–2 00101 10101 000–0 00101 100–1 000–1 000–1 000–1 000–0 00101 300–0 00101 00101 000–2 000–2 000–2 00111 000–2 001?0 00101 100–0 300–0 000–2 000–1 000–1 010–1 000–2 100–0 000–1 000–1 00101 00101 000–1 00101 000–1 00101 11011 00012 00011 10011 00011 00011 00011 00111 10011 00011 11011 31011 01111 20011 00010 10011 10011 10011 00011 21011 11011 31011 00111 10011 00011 00011 20011 10011 20111 31011 30011 10011 21011 10111 20011 11011 10111 10111 01011 20011 10111 20100 10110 11111 12110 111?? 10101 12100 110–1 10100 12100 10112 110–2 10101 001?? 10100 12101 12101 11112 11100 10101 13111 10??? 12101 10100 00100 10110 11112 10100 10111 13102 11100 11110 130–0 101?? 12110 130–2 12102 10100 10101 00001 10000 10001 10001 10000 10001 10000 00111 –0001 00000 10011 10010 –0100 10010 10000 10000 10001 10001 10001 10001 10010 10001 11010 10000 10000 10010 11011 11011 00010 10010 10001 10001 10001 –0001 10010 10010 –1111 00111 11011 10010 10000 00000 20000 20111 00000 00000 00000 20111 20000 00000 01010 00000 00111 01011 01010 00001 00000 00000 10000 20001 00001 30001 00001 00000 00000 01011 00001 00001 00111 01011 30001 10001 20001 31011 00001 00001 30110 20110 00111 00111 00101 11000 11000 01000 00000 11000 00000 01000 11000 11000 00111 00000 01000 01000 00000 11000 00110 00110 00110 11000 11101 11000 11000 00110 00100 11000 11000 01000 01000 11100 01000 11000 11000 01000 11000 11101 00100 00100 01000 01000 00010 00010 00000 00000 00000 00000 00000 11100 10000 00000 00000 00000 00000 00000 00000 00000 00000 00000 00000 00000 00110 00000 00010 00000 00000 00010 00000 00000 00000 00000 00000 00010 00000 11000 00010 00010 00000 00000 00000 00000 84 TABLE 2. (cont.) Data matrix used for the cladistic analysis – = inapplicable, ? = unavailable character state Taxa Character numbers and states 1 11111 11112 22222 22223 33333 33334 44444 12345 67890 12345 67890 12345 67890 12345 67890 12345 Buellia penichra Buellia pharcidia Buellia prospersa Buellia proximata Buellia pullata Buellia pulverentula Buellia regineae Buellia romoletia Buellia rubroreagens Buellia ryanii Buellia scheideggeriana Buellia sequax Buellia sheardii Buellia sorediata Buellia spuria Buellia stellulata Buellia subaethalea Buellia subalbula Buellia subdisciformis Buellia subdispersa Buellia submuriformis Buellia tergua Buellia terricola Buellia tesserata Buellia tombadorensis Buellia trachyspora Buellia triphragmoides Buellia triseptata Buellia tyrolensis Buellia uberior Buellia venusta Buellia vernicoma Buellia vilis Dimelaena oreina Dimelaena radiata Diploicia canescens Hafellia leptoclinoides Hafellia parastata Rinodina bischoffii Rinodina confragosa 01000 01000 11200 01?00 10200 0002– 11000 00000 01?00 11001 10?2– 10000 11100 00?00 11000 11000 11000 11000 11100 10?00 00?00 10000 20?00 11000 00000 11100 00000 00011 11000 10020 11100 30?00 10000 10001 11001 30001 11000 01?00 11000 11000 10001 00011 00001 00001 00000 ––010 11002 10000 00001 11000 ––010 01000 11001 10101 11000 10000 10000 00010 00001 11010 00001 00000 10011 10000 00001 00002 10001 1?000 10000 11000 00011 10000 01001 11000 11000 10011 10001 1?002 00011 10002 000–0 10101 000–0 000–1 000–0 00100 000–1 000–0 000–1 100–0 10100 000–0 000–1 000–1 100–0 100–0 000–1 00101 000–1 10100 000–1 000–1 00101 10100 00111 000–2 00111 000–0 100–1 100–0 10100 00110 000–1 11101 310–1 100–1 000–1 000–1 010–0 010–0 10111 00011 00011 10011 00011 10011 20111 10011 10011 00011 01011 00011 20011 10011 00011 00011 00011 00011 11011 00011 00011 00011 10011 00011 10011 40011 10011 10011 00011 00011 00011 11011 51010 30011 30011 10011 11111 11111 00100 00000 120–0 10110 10101 11100 10100 10110 10101 11101 11110 101?? 10110 10100 101?? 01101 10100 10101 11111 10100 101?? 10110 13111 10100 12101 10101 11110 12112 11101 10101 10100 10100 10110 10100 00100 11101 11101 10100 100–2 13101 01101 01110 –0001 10001 10010 11011 10010 10001 00110 10001 11011 10000 10001 10000 00000 10010 10000 10000 10010 00000 10010 10001 10001 10000 10001 10000 11011 10001 10001 10001 00000 10000 10001 10001 00000 00000 10000 11010 –0100 00110 00010 11010 20001 10000 00001 00111 00000 00000 01111 00000 00001 00000 10000 00000 00001 00101 00001 01011 00000 00000 01011 00000 20111 00000 00001 01011 00000 00000 00000 00001 01010 01010 00000 00000 00000 01011 01011 00111 00001 00111 00000 00010 01000 00000 11100 01000 00000 00000 01000 00110 11000 00000 00000 11000 11000 00000 11000 01000 11100 11000 11000 00000 01000 00000 11111 11000 00101 00100 00101 01000 11000 00000 11000 00101 00000 11000 11000 01100 01000 01000 00000 10000 11000 00000 00000 00000 00000 00000 00000 00000 00001 00010 00000 00000 00010 00000 00010 00010 00010 00010 00000 00000 00000 00010 00000 00000 00001 00000 00000 00000 00000 00010 00000 00000 00000 10000 00000 00000 11000 00000 00000 00000 85 Discussion: Segregates from the Genus Buellia and Taxonomic Implications The low levels of bootstrap support and the large number of equally most parsimonious trees of 385 steps are not surprising given the high number of taxa examined and the low number of characters available for comparison. Nordin (2000) also argued that the poor branch support of his analysis was a result of the large number of taxa and the relatively small number of characters. Only eight additional characters were added here and three of Nordin’s characters were not used. The cladistic analyses had several objectives: (1) to test Nordin's (2000) conclusion that species with pluriseptate spores do not form a monophyletic group, (2) to examine if corticolous taxa are well separated from saxicolous ones, (3) to examine if any parasitic taxa form a distinct group, (4) to examine if species with filiform conidia form a monophyletic group, and (5) to explore possible consequences of accepting several generic segregates of Buellia s.l. based only on classical evidence. Nordin's (2000) conclusion that species with pluriseptate spores are not monophyletic was well supported by his original analysis. Even if more species with one-septate spores are added to his data set, species with pluriseptate spores do not form a monophyletic group. However, some groups with predominantly one-septate spores can be found and at least one poorly resolved group of species includes only species with pluriseptate spores. Diploicia canescens is basal to this group and both Nordin (2000) and Giralt (2000) suggested that this genus might also include non-placodioid species. Particularly interesting is the B. alboatra-group, the core group of Diplotomma. In the strict consensus tree Buellia amabilis, a species with one-septate spores, can be found 86 within this group. This is additional evidence that this species could be included within Diplotomma s.str. (Bungartz & Nash III 2004a, Publication 4) Marbach (2000) suggested that his classification should have little or no impact on the classification of saxicolous taxa. This statement cannot be confirmed by the cladistic analysis presented here. Saxicolous and corticolous taxa are intermixed throughout the consensus tree and corticolous taxa do not form distinct monophyletic groups. Several corticolous species treated by Marbach (2000) within separate genera are not isolated from other groups. Although only a few corticolous species with one-septate spores were included in the analysis, it is unlikely that more distinct groups of corticolous species would have been supported. To the contrary, additional taxa would probably have contributed to even less resolution. Only molecular characters will ultimately provide an adequate number of characters to resolve sufficiently relationships to evaluate Marbach's (2000) assessment. Nevertheless, it seems likely that the classification presented by Marbach (2000) potentially has considerable taxonomic importance even for the saxicolous taxa. Only a few taxa included in the analysis are obligate or facultative parasites. None of these taxa form a distinct group. Symbiotic interactions are generally very complex and several different interactions have been described (Rambold & Triebel 1992). Lichens are polyphyletic (Gargas et al. 1995; Lutzoni et al. 2001), and lichen parasites have evolved independently on several occasions. It is therefore not justified to isolate some parasitic species from Buellia s.l. 87 Three species included in the analysis have filiform conidia. They do not form a monophyletic group in any of the equally most parsimonious trees. Buellia prospersa, a species with filiform conidia is part of a group of species, which all contain xanthones but otherwise have bacilliform conidia. Buellia pullata is another species with filiform conidia. It occurs in a different clade from B. prospersa, as part of a poorly resolved polytomy, which includes B. coniops, the type of Amandinea, together with several other species, which all have bacilliform conidia. The following paragraphs discuss genera segregated from Buellia s.l. It is evident from this analysis, that the classical data used here is insufficient to provide support for most of these segregates. Nevertheless, some groups can be recognized here, as well as in the analysis previously presented by Nordin (2000). The taxonomic implications of these segregates are briefly discussed in the context of the saxicolous species treated in this dissertation. In addition, Marbach (2000) accepts several other generic segregates of Buellia s.l. Most of these segregates are problematic (Eriksson et al. 2002b), but only a few currently include both saxicolous and corticolous species. Amandinea M. Choisy ex Scheid. & M. Mayrhofer The genus Amandinea was introduced but not validly described by Choisy (1950). Scheidegger (1993) provided the first valid description, and emphasized the filiform conidia as the main diagnostic character. Although the type species Amandinea coniops is saxicolous, many corticolous taxa have also been transferred into Amandinea (Sheard & May 1997; Marbach 2000; Søchting et al. 2004). Species with lecideine apothecia initially described as Buellia as well as species with lecanorine apothecia, originally 88 → FIGURE 13. Strict consensus tree of one thousand equally most parsimonious trees (bold numbers indicate bootstrap values). 89 90 described as Rinodina have been transferred into Amandinea. As a result, Amandinea now includes many species which differ widely in their apothecial anatomy, thallus chemistry, spore structure, thallus morphology and substrate. This amalgam of species based solely on conidial length, is not very convincing. A large variation of conidial length can generally be observed in Buellia s.l. and conidiophores and pycnidia show no significant differences (Bungartz et al. 2004b, Publication 5). The phylogenetic analysis presented here does not support Amandinea. At least B. prospersa appears to be more closely related to species with bacilliform conidia containing xanthones than to species with filiform conidia and no secondary lichen metabolites. Marbach (2000) described the new genus Fluctua based on the single species Fluctua megapotamica (Malme) Marbach. The genus is characterized by filiform conidia and appears not distinctly different from Amandinea (Eriksson et al. 2002b). In a cladistic analysis of classical characters, Scheidegger et al. (2001) examined evolutionary trends in the Physciaceae and argued that Amandinea was well supported. However, only three species were included in their analysis. Helms (2003) demonstrated that Amandinea belongs to the Buellia-clade, but only two species were included in his molecular analysis. The genus concept of Amandinea needs to be revised and only species closely related to the type Amandinea coniops should be included. Aplotomma A. Massal. ex Beltr. Nordin (2000) demonstrated that this segregate of Diplotomma is not closely related to the Buellia alboatra-group, and therefore was not synonymous with Diplotomma s.str. Nevertheless, Nordin (2000) did not accept Aplotomma because his phylogenetic analysis 91 did not provide enough resolution to segregate the type Buellia (= Aplotomma) griseovirens from other species of Buellia s.l. In the analysis presented here Buellia griseovirens can be found in a group of species with pluriseptate spores, including the genus Diploicia. Baculifera Marbach & Kalb Marbach (2000) described this new genus primarily based on bacilliform conidia, but also emphasized the presence of norstictic acid within the exciple, its black prothallus and its hymenium, which is not inspersed with oil droplets. All these characters are common across Buellia s.l. Norstictic acid is indeed one of the most common lichen substances in Buellia s.l. and the presence of bacilliform conidia has been used to segregate the genus from Amandinea. Marbach (2000) mentions that many, but not all Baculifera species, have ascospores with slight lateral wall thickenings. Lateral thickenings are also diagnostic for Hafellia, which, according to Marbach (2000), has shorter and broader conidia, more strongly thickened lateral spore walls, a hymenium inspersed with oil droplets, and lacks a black prothallus. Marbach (2000) argues that both Hafellia and Baculifera can usually be distinguished but that H. pruinosa is problematic because no conidia of that species were found. The variation in conidia – short and thick bacilliform conidia vs. slightly longer, less thickened conidia – as the most diagnostic character to distinguish the two genera seems rather daring. It is incorrect to suggest that Hafellia lacks a black prothallus. Scheidegger (1993) transferred Buellia leptoclinoides, a species with a very distinct black prothallus, into Hafellia because of the inspersed hymenium and the diagnostic lateral 92 wall thickenings of the ascospores. Buellia regineae (Bungartz 2004, Publication 8) also has a distinct black prothallus, but inconspicuous lateral wall thickenings and an oil inspersed hymenium. A black prothallus can even be found in some specimens of H. parastata, the only corticolous species included here. The North American species Buellia imshaugiana was included by Marbach (2000) in Baculifera. The species has indistinct lateral spore wall thickenings, but groups more closely with B. erubescens, not with any species of Hafellia. Buellia erubescens and B. imshaugiana are both corticolous. In the phylogenetic analysis they are part of a group of species, which are all saxicolous, including B. aethalea, a species accepted by Marbach (2000) as the type of Buellia s.str. Further studies are necessary to evaluate if Baculifera must be recognized as a distinct genus. Dimelaena Norman The genus is closely related to Buellia because of its thin-walled ascospores, frequently dark hypothecium and its Bacidia-type ascus. The only two species included in the analysis formed one of the very few groups supported by non-parametric bootstrapping. The main character of the genus is its distinctly lobate thallus. It has been suggested that Dimelaena radiata is very closely related to Buellia tesserata (Rico et al. 2003). Both species share the same thallus chemistry, have apothecia with almost identical anatomy and very similar spores. However, Buellia tesserata does not have a lobate thallus margin and can only be included within Dimelaena, if the genus concept is emended to accommodate non-lobate taxa. Such a change is problematic because the phylogenetic analysis suggests that Dimelaena would then have to include B. tyrolensis 93 or even B. stellulata, a species with many characteristics of the B. aethalea-group (Bungartz & Nash 2004c, Publication 6). Diploicia A. Massal. The genus was reintroduced by Poelt (1973) and subsequently widely accepted. Species commonly included have pluriseptate spores and a distinctly lobate thallus. Nordin (2000) suggested that the genus may be too narrowly circumscribed, and possibly also includes species not delimited by distinct lobes. Giralt (2000) studied Rinodina kalbii and R. ericina and also suggested that those two species might better be included within Diploicia rather than Rinodina. Molina et al. (2002) demonstrated that the species included in Diploicia were closely related to Diplotomma based on rDNA ITS sequence data. They suggested that the two genera should therefore be united. Helms (2003), did not agree with that assessment and suggested that ITS sequences and spore characteristics of Diploicia were sufficiently different to distinguish both genera. The cladistic analysis by Nordin (2000) shows Diploicia as part of a group with similar, pluriseptate ascospores separated from Diplotomma s.str. (Buellia alboatra-group). That result corresponds with the phylogenetic analysis presented here. Diplotomma Flot. Nordin (2000) demonstrated convincingly that species of Buellia with pluriseptate spores do not form a monophylletic group and must not generally be included in Diplotomma. He did not formally accept the genus but emphasized that a small subset of species (the Buellia alboatra-group) could be recognized as Diplotomma s.str. This 94 Buellia alboatra-group is characterized by the presence of calcium oxalates in the thallus, often pruinose apothecia and a thick perispore. Bungartz & Nash (2004a, Publication 4) examined two species with similar characteristics but one-septate ascospores and concluded that Buellia subalbula cannot be included in Diplotomma because it is characterized by a thin perispore. However, a thick perispore was observed in B. amabilis, which therefore could be included Diplotomma s.str. even though it has one-septate ascospores. This assessment is also supported by the phylogenetic analysis. Gassicurtia Fée The name Buellia De Not. was conserved against Gassicurtia Fée, when Aptroot (1987) rediscovered that this older name Gassicurtia would have had taxonomic priority. Marbach (2000) resurrected the genus, arguing that Gassicurtia was a distinct group of species not included in Buellia. Following his concept, the conservation of Buellia in the botanical code (Greuter et al. 2000) would be superfluous. Marbach (2000) suggested that granulose to subgranulose thalli and the presence of the orange-red, K+ purple pigments in the thallus and apothecia are the most diagnostic characters of Gassicurtia. Nordin (2000) found several species with pluriseptate spores clustering within Gassicurtia, including one species treated by Marbach (2000) in the genus Mattickiolichen. The consensus tree presented here, suggests that Buellia concinna might be part of that group. The species has a granular thallus and contains xanthones but lacks the characteristic orange red, K+ purple pigment. Buellia capitis-regum is a common coastal species in the Sonoran Desert Region. The species is characterized by an orange, K+ purple medulla, but it does not group with 95 Gassicurtia. Nordin (2000) and the phylogeny presented are both consistent with the interpretation that B. capitis-regum might be part of Hafellia s.l. Grube et al. (2004) describe two new saxicolous species of Buellia with orange red pigments from Australia. They emphasized that these species do not share other characters with Gassicurtia although the two species were not included in the phylogenetic analysis. Hafellia Kalb, H. Mayrhofer & Scheid. Hafellia is characterized by apothecia with an oil inspersed hymenium and spores with lateral wall thickenings. Many species of Hafellia are also characterized by diploicin and related substances. Marbach (2000) reports traces of fulgidin, a substance closely related to diploicin, from B. disciformis. Hafellia leptoclinoides does not contain diploicin, but instead is characterized by the rare secondary metabolite placodiolic acid (Scheidegger 1993). Nordin (2000) reported that both Hafellia leptoclinoides and B. disciformis lacked a perispore. Most other species appear to have a very thin perispore, for example Hafellia parastata (Nordin 2000). Furthermore, B. oidalea and B. oidaliella are also characterized by a thin perispore, contain species diploicin and group with Hafellia even though they have pluriseptate spores (Nordin 2000). Buellia regineae is a closely related species from the Sonoran Desert Region, also characterized by diploicin an a thin perispore (Bungartz 2004, Publication 8). Once the nomenclatural problems of Hafellia are resolved, all these species might be included within the genus. This combination of characters is quite unique and the species included in Hafellia therefore appear closely related. The genus could be accepted but the presently valid 96 botanical code (Greuter et al. 2000) lists Buellia disciformis as the type of Buellia. As a consequence the genus name Hafellia must be treated as a synonym of Buellia s.str., if the inclusion of B. subdisciformis within Hafellia by Marbach (2000) is accepted. To avoid this conflict Moberg et al. (1999) suggested changing the listed type of Buellia but this proposal is not currently accepted. Recent molecular data reveal that B. disciformis may not be closely related to Hafellia (Mayrhofer, pers. comm.). Thus it may not be necessary to change the listed type of Buellia for the sake of Hafellia, but the classic concept of Hafellia (oil inspersion, presence of diploicin and related substances, lateral spore wall thickenings, perispore thin or absent) appears much less distinct if B. disciformis is not included. From the Sonoran Desert Region, B. regineae is the only saxicolous species with one-septate spores that shows distinct affinities to Hafellia (Bungartz 2004, Publication 8). However, the species is described as a Buellia until the uncertain taxonomic status of Hafellia is resolved. Hafellia originally included only species with one-septate ascospores, but Marbach (2000) transferred several species with pluriseptate spores into Hafellia, and that concept is supported by Nordin (2000) and the phylogenetic analysis presented here. Karschia Körb. Hafellner (1979) revised a large group of lichenicolous fungi with brown, one-septate ascospores, formally included in Karschia s.l. Most taxa once treated in Karschia s.l. belong to the Dothideales, including Karschia s.str. These genera are all obligate lichen parasites, and are very distantly related to Buellia, which belongs to the Lecanorales. Only a small group of the specimens examined by Hafellner (1979) were retained within 97 Buellia. These species, including B. badia, are parasitic at least during their juvenile stages. Marbach (2000) suggested that B. badia was better placed in Monerolechia, but that genus cannot be accepted (see the discussion of below). Mattickiolichen Tomas. & Cif. Nordin (2000) explains that the genus name is based on the odd idea of giving separate names to the lichen and its fungal components (Tomaselli & Ciferri 1952; a proposal criticized by Santesson 1954). Marbach (2000), nevertheless, accepted the genus arguing that no other species of Buellia s.l. have Umbilicaria-type pycnidia. That assessment is not supported by the microscopic studies presented here. All species with bacilliform conidia have pycnidia, that are very similar, if not identical with the Umbilicaria-type. In the phylogenetic analysis Buellia triphragmoides, the type of Mattickiolichen, clades with several other species including some species of Gassicurtia sensu Marbach (2000). This group was also supported by Nordin's (2000) analysis. Melanaspicilia Vain. Marbach (2000, p. 342) selected Melanaspicilia sororia (Th. Fr.) Vain. as lectotype of Melanaspicilia. Buellia sororia Th. Fr., the basionym of Melanaspicilia sororia, is conspecific with Buellia aethalea. Melanaspicilia can therefore only be accepted if the proposal by Moberg et al. (1999) to change the listed type of Buellia to Buellia aethalea is rejected. Moberg et al. (1999) argued that the listed type of B. disciformis belongs to Hafellia. Marbach (2000) assumed that their proposal had been accepted, and included B. disciformis within Hafellia. According to Mayrhofer (pers. comm.) preliminary 98 molecular evidence indicates that B. disciformis may not be closely related to Hafellia, even though it shares many characters with other species in this genus. However, under the currently valid code (Greuter et al. 2000) the classical concept of Buellia with the B. aethalea-group as the core group can only be maintained if molecular evidence clearly demonstrates that B. disciformis is indeed closely related to that group. This appears very unlikely, and the genus Melanaspicilia might have to be used for species in the Buellia aethalea-group, if B. disciformis remains the listed type of Buellia s.str. In the strict consensus tree species of the Buellia aethalea-group (“Melanaspicilia”) are not well resolved. They are nested within a clade which includes Dimelaena s.l., “Baculifera” and a group of species with xanthones. Monerolechia Trevis. In the Sonoran Desert Buellia badia grows as a facultative juvenile parasite. The species was included by Marbach (2000) in Monerolechia. In the same treatment, he included B. turgescens within Amandinea, even though that taxon has bacilliform conidia and must be regarded as a synonym of Buellia badia (Bungartz & Nash 2004b, Publication 2). The suggestion that parasitic species have to be segregated from Buellia is not convincing, especially if some of the species are only facultative lichenicolous during early stages of their development. Marbach (2000) accepts other parasitic species in some of his genera (e.g. Gassicurtia bellardii). He lists the type species of Monerolechia as M. bayrhofferi, which he considers a synonym of Buellia badia. Thereby, he creates in effect 99 the new combination M. badia (Fr.) Marbach, even though he does not list this name (Article 33 ICBN, Greuter et al. 2000). Hafellner (1979) treated Monerolechia as a synonym of Buellia and emphasized that there was no justification to segregate the parasitic taxa. More recently Hafellner (2004) discussed that many crustose Physciaceae showed independent tendencies to become lichenicolous. Because of their filiform conidia he transferred several obligate parasymbionts into Amandinea. This transfer may be problematic because the genus concept of Amandinea has to be re-evaluated. However, the species were not separated from Buellia because of their parasitism. Indeed, in the same publication Hafellner (2004) did not accept Monerolechia, but instead included B. badia within Buellia. In the same Festschrift as the paper published by Hafellner, Kalb (2004) published the new combination Monerolechia badia (Fr.) Kalb, and argued that the genus was well supported both by a different ascus-type and an exciple structure different from Buellia s.str. However, his new combination must not be accepted for several reasons: (1) The combination Monerolechia badia (Fr.) Kalb is taxonomically not valid. It must instead be cited as Monerolechia badia (Fr.) Marbach. Article 33.1. of the botanical code (Greuter et al. 2000) states: “A combination (autonyms excepted) is not validly published unless the author definitely associates the final epithet with the name of the genus or species, or with its abbreviation.” Marbach (2000) clearly indicated that he recognized B. badia as a synonym of B. bayrhofferi, the type of Monerolechia, which he accepts as a valid genus. His treatment of Monerolechia therefore agrees with Art. 33.1., 100 i.e. Marbach (2000) definitely associated the final epithet “badia” with the genus Monerolechia. (2) The neotype material of B. badia was not studied by Kalb (2004). Only a single specimen from Australia is listed. This specimen may not belong to B. badia, if it indeed displays a different ascus type than the neotype selected by Foucard et al. (2002) [also see the next argument (3)]. (3) The ascus of the neotype of B. badia selected by Foucard et al. (2002) is identical with the Bacidia-type, not the Lecanora-type. Kalb (2004) argued that B. badia had an ascus type similar to Physcia tenella as depicted by Marbach (2000, p. 28, Fig. 10D), but that statement is not supported by the type material. Buellia badia has a broad, I– inner tholus with almost parallel, I+ blue flanks. The flanks merge at the tip, forming a thin, but distinctly I+ blue band. In contrast, the central, I– part of the tholus in P. tenella widens at the apex and the I+ blue flanks do not merge at the tip. Thus, the ascus of B. badia belongs to the Bacidia-type, whereas P. tenella belongs to the Lecanora-type. This assessment is well supported by molecular phylogeny, which includes Buellia as part of the Buellia-clade and Physcia as part of the Rinodina-clade (Helms 2003). (4) The proper exciple of Buellia aethalea (referred to as a parathecium by Kalb 2004) is reduced to a few hyphae similar to the paraphyses, and therefore is much less distinctly developed than the exciple of Buellia badia. The excipular hyphae of B. badia, nevertheless, have the same structure and pigmentation as excipular hyphae of B. aethalea. In fact, the exciple structure of immersed, immature apothecia of B. badia is identical with the exciple structure of B. aethalea. A reduced exciple in B. aethalea is 101 clearly the result of an apothecial ontogeny, where the exciple is less developed because the apothecia remain immersed. Buellia spuria, a species closely related to B. aethalea, initially also has a very narrow exciple. When the apothecia of B. spuria emerge, the exciple expands and the structure becomes identical to the exciple of mature, sessile apothecia of B. badia. Thus, both B. aethalea and B. badia must be assigned to the same aethalea-type sensu Scheidegger (1993). The phylogenetic position of B. badia is currently very poorly resolved. The strict consensus tree shows the species together with the non-parasitic, corticolous B. triseptata, which has pluriseptate spores, but this group is not well supported. It is a sister group of two clades: one that includes a group with “Mattickiolichen” and “Gassicurtia”, and another one of terricolous species. The other large clade includes species of the B. dispersa-group, Hafellia s.l. and Diploicia s.l. Rinodina (Ach.) Gray Several molecular studies confirm that the Physciaceae can generally be segregated into two large clades with Rinodina s.l. as a distinct sister group of Buellia s.l. (Wedin et al. 2000; Wedin et al. 2002; Wedin & Grube 2002; Helms 2003; Helms et al. 2003). Relatively low bootstrap support for Rinodina as an outgroup is the result of the few classical data available and does not contradict this assessment. The taxonomy of Rinodina s.l. is as complex and poorly resolved as the taxonomy of Buellia s.l. The genus Rinodina s.l. is not monophyletic (Helms 2003). Several segregates have been suggested but these are not generally accepted. Marbach (2000) described Sculptolumina as a new genus and included only two species, but Aptroot (2002a) argued 102 that the genus could not be distinguished from the Rinodina oxydata-group. The type Sculptolumina japonica (Tuck.) Marbach is synonymous with Rinodina atrofusca (Vain.) Aptroot (Aptroot 2002a). The species epithet japonica has taxonomic priority and Aptroot (2002b) suggested the new combination, before he became aware of the synonymy (Aptroot, pers. comm.). Sculptolumina was not included in the phylogenetic analysis because it was not an objective of this analysis to examine the phyologeny of Rinodina s.l. Tetramelas Norman Marbach (2000) reintroduced this genus to accommodate terricolous species with oneseptate ascospores. Trinkaus et al. (2001) argued that molecular data do not support a general segregation of terricolous taxa (the Buellia epigaea-group). Nordin's (2000) phylogeny grouped terricolous species with pluriseptate spores group with species with one-septate spores. He suggested that Tetramelas might be used to describe a group of terricolous, arctic-alpine species with large ascospores and the xanthone 6-Omethylarthothelin, and that group is also supported here. However, terricolous species from arid or semi-arid regions like the Sonoran Desert or Australia (Trinkaus et al. 2001), were not included in the analysis and these species or may not belong to the group. Buellia species with xanthones The consensus tree presented here unites several species with xanthones within a clade. It has not yet been suggested to segregate species of Buellia with xanthones as a separate genus. These species are morphologically and anatomically quite different, and 103 it is doubtful that they are indeed closely related. Buellia subdisciformis is included in the group, even though it does not contain xanthones. The exuberant, chalky thallus of B. navajoensis is very different from thalli of the other species. Buellia mamillana has a unique exciple type not found in any of the other species. Several other species of Buellia also contain xanthones, but are not included in this group (e.g., B. trachyspora, B. concinna, B. oidalea, B. aeruginosa etc.). Conclusions Many taxonomic questions could not be addressed with the phylogenetic analysis presented here. For example, B. vilis or B. trachyspora are very isolated even in a broad concept of the genus. Their isolated position is, however, not evident in any of the trees, even though they were both assigned a separate exciple type. Reducing the complex exciple structures to a single character is clearly inadequate. It might have been more appropriate to distinguish several additional characters such as the diameter of the hyphae, hyphal orientation within the exciple, wall thickness, amount of pigmentation, shape of cells etc. All of these characters could have been examined separately for the inner, central and outer exciple. However, this information would have required additional research and data were not available for all species included in the analysis. Even if the data set could have been expanded, adding a few more characters may not have been insufficient to achieve a better resolution. Nordin (2000, p. 40) argued: “The relatively high number of species and the relatively few characters available made it necessary to minimize the number of species with 1-septate spores.” It is therefore quite a 104 surprise that most of his groups could be confirmed even though almost twice as many species were included in this analysis. 105 PUBLICATIONS PUBLICATION 1 (Bibliotheca Lichenologica 82, 2002): Buellia dispersa A. Massal., a variable lichen species from semi-arid to arid environments of North America and Europe FRANK BUNGARTZ and THOMAS H. NASH III Department of Plant Biology, Arizona State University P.O. Box 87 1601, Tempe, AZ 85287-1601, U.S.A. please address correspondence to: frank.bungartz@asu.edu CHRISTOPH SCHEIDEGGER Swiss Federal Institute for Forest, Snow and Landscape Research, WSL, CH-8903 Birmensdorf, Switzerland Abstract. The North American taxa Buellia retrovertens, B. blumeri, B. tucsonensis and B. cinereoglauca are synonymized with B. dispersa. The species was originally described from Europe where it occurs in the Mediterranean and the inner alpine dry valleys. In North America the species grows in arid to semi-arid environments of the southwestern United States and northern Mexico. It is abundant in the Sonoran Desert Region. North American specimens are compared with specimens from France, Greece, Italy and Switzerland. The specimens show a very variable thallus morphology and three morphotypes can be recognized according to thallus growth, color and pruinosity. However, these morphotypes do not correlate with other characters. All specimens are 106 characterized by the presence of atranorin, 2'-O-methylperlatolic, +/- confluentic acid and rarely an unknown substance. The thalli have lecideine apothecia with a dispersa-type exciple. The hypothecium is dark reddish brown and extends into the inner exciple. During a brief phase of their ontogeny ascospores show a median thickening. A microrugulate ornamentation can be observed only during the last stages of spore development. Buellia dispersa is a morphologically very diverse lichen species described by Abramo Massalongo in 1853. In his monograph of the European species of Buellia s.l. with one septate ascospores, Scheidegger (1993) ascribes this variability to habitat variation and consequently synonymizes B. italica var. tumida, Lecidea squamulata, Lecidea dispersa var. subeffigurans, B. tergestina, B. duartei and B. subsquamosa non Steiner with Buellia dispersa. Several taxa with close affinities to the European material of B. dispersa have been described from specimens found in the southwestern United States and northern Mexico, especially the Sonoran Desert Region. Imshaug (1951) suggested reducing these taxa - B. blumeri, B. tucsonensis and B. cinereoglauca - to synonymy with B. retrovertens. Even though his thesis was never validly published, B. retrovertens was subsequently accepted by most North American lichenologists as the correct name for the morphologically diverse group. Nevertheless the taxon remains poorly understood and the immense variation of thallus morphology causes much confusion. Type specimens as well as other material of the species synonymized by Imshaug (1951) have not been studied recently, 107 and it is not surprising that the similarity of the North American material with B. dispersa has so far been overlooked. Thallus morphology of B. dispersa is extremely variable (Figs. 14 and 15), especially in the North American specimens. Imshaug (1951, p. 126) mentions difficulties finding specific characters to distinguish distinct separate groups: “...I have tried to find some characters that could be used to divide this variable population into two or more homogeneous groups. The apothecia, however, are quite uniform and offer no solution. The thallus characters, on the other hand, are far too variable to be used in species delimitation. ...” In his revision of the European taxa, Scheidegger (1993) comes to similar conclusions regarding thallus variability in B. dispersa. He attributes the variation mainly to substrate differences and emphasizes the uniformity in excipular structure. We have studied material of the taxa which Imshaug (1951) suggested to synonymize as B. retrovertens. This material from the southwestern United States and northern Mexico, especially the Sonoran Desert Region, was compared with selected specimens of B. dispersa from France (East Pyrenees), Greece (Sterea Ellas), Italy (Sardinia, Trieste region) and Switzerland (Wallis). As part of a larger study of Buellia species from the Sonoran Desert Region we have also been able to examine previously unidentified specimens of Buellia dispersa from various collections. 108 MATERIALS AND METHODS The following material was examined: FRANCE: Scheidegger Inv.Nr.7547, 8251 (herb. Scheidegger). GREECE: Scheidegger Inv.Nr.12477 (herb. Scheidegger). ITALY: Scheidegger Inv.Nr. 11136 (herb. Scheidegger), Krypt. Exs. Vindob. 85 (US, S). MEXICO: Moberg 8646, 8902 (UPS); Nash 8951, 9022, 11117, 12083, 12435, 12477, 12746, 12889, 13818, 13836, 13885, 14082, 25595, 25660, 26075, 26092, 26139, 29579, 30278, 30354, 36101, 37950, 37951, 38270, 38446, 38457, 39068, 39663, 39982, 40069, 40070 (ASU); Ryan 21595, 21643 (ASU); Scheidegger Field No. 15, 123, 128, 130, 139, 141, 144, 149, 160, 164, 165, 260; Inv. Nr. 10421 (herb. Scheidegger); Sipman 24947, 24965, 24990, 25026, 25125 (B) Weber L-33667 (COLO); Wetmore 72077 (MIN). SWITZERLAND: Scheidegger Inv.Nr. 8580. U.S.A.: Arsène 19727 (US), Bessey s.n. (leg. 1908, DS, now integrated with CAS, accession no. 686248), Biringer 7 (ASU), Blumer 6777 (W, MICH), 6795 (MICH); Boykin 2689 (ASU); Brandegee in Herb. Sprague (ex Tuckerman sheet no. 3281, FH); Bratt 4960, 10164, 10300 (SBBG); Bungartz 1427, 1488(herb. Bungartz); Darrow 682a, 1305, 1409, 1878, 5138a, 5138b, 5139, 4307, 5137 (ASU); Gebauer s.n. (ASU), Hasse Exs. 81 (ASU, DS); Lane 172, 1878 (ASU), Marsh 118, 119, 394 (ASU); Nash 1878, 3694, 4315, 4317, 5070, 6009, 6010, 6011, 6020, 6049, 6146, 6158, 6192, 6334, 6480, 6578, 6851, 7801, 7818, 8463, 8532, 9155, 9222, 9605, 9837, 9946, 9966, 10443, 10754, 15312, 15314, 15985, 15998, 16084, 16312, 22427, 22984, 25273, 26075, 27395, 30660, 30676, 30712, 35138, 35139, 35162, 40268, 40568, 40592, 40593, 40605 (ASU); Printzen Field No. 24 (herb. Printzen); Ryan 15148, 15818, 16048, 16098, 19163, 19191, 20536, 30880, 30883, 109 32872 (ASU); Scheidegger Field No. 13, 19, 22, 26 (herb. Scheidegger); Sigal 39 (ASU); Sundell 15 (ASU); Weber S-8832, L-36718, L-36732, L-78824 (COLO); Wetmore 18289, 55371 (MIN). The specimen distribution map (Fig. 7) was generated using ArcView GIS 3.2. Photographic plates were assembled and edited in Adobe Photoshop 6.0 and printed on a CODONICS NP-1660M high resolution printer. Thallus variability of the specimens was compared with a dissecting microscope and documented by conventional photography. The cortex structure was studied from handcut thallus sections and 20-30 µm thick freezing microtome sections. Additionally some specimens were air-dried, sputter coated with gold and viewed with a Jeol JSM-840A scanning electron microscope at accelerating voltages between 7 and 12 kV. Specimens were spot tested and lichen compounds analyzed using standardized thin layer chromatography (Culberson & Kristinsson 1970; Culberson & Johnson 1982; White & James 1985). Plates were interpreted with WINTABOLITES (Mietzsch et al. 1994) and scanned into the computer for permanent record. Excipular anatomy was studied with conventional light microscopy, phase contrast and differential interference contrast. Color reactions of the apothecial pigments with 10% KOH and conc. HNO3 were observed from hand-cut sections. Excipular anatomy was studied from 11 µm thin sections cut with a Reichert HistoStat 855 freezing microtome. In addition, scanning electron microscopy (SEM) allowed the study of the three dimensional structure within the plectenchyma. Thick median sections of apothecia from both European and North American specimens were incubated overnight at 34 °C in 110 a saturated solution of commercially available “Ariel” washing powder (HONEGGER 1985). After repeated, thorough rinsing in distilled water, sections were fixed for 2-4 hrs in 1% aqueous OsO4, dehydrated in a 40, 60, 80, 100% series of acetone, critically point dried, mounted on aluminum stubs, and sputter coated with gold. They were then examined with a Jeol JSM-840A scanning microscope at accelerating voltages between 7 and 12 kV. SEM also helped to distinguish spore surface ornamentation but it was necessary to compare these observations with light microscopy ones. Ascus apical structures were examined from squash preparations treated with 10% KOH and subsequently Lugol’s iodine. The iodine reaction is more pronounced, however, if the solution is made with 1g of iodine crystals rather than 0.5 g. Spore ultrastructure was studied using transmission electron microscopy (TEM). Apart from some modifications, specimen preparation followed Nordin (1997). Apothecia were fixed for 48 hrs in 2% 0.05M glutaraldehyde in pH 7.2 sodium cacodylate buffer. Secondary fixation was 2 hrs 15 min in 1% OsO4 in pH 7.2 sodium cacodylate followed by en-bloc staining for 2 hrs in 0.5% aqueous uranyl acetate. Between these steps each fixative as well as the en-bloc staining solution was carefully washed out with at least 3 changes of Barnstead water, the last change overnight. Finally specimens were dehydrated in a series of 20, 40, 60, 80, two changes of 100% ethanol and subsequently two changes of 100% acetone. They were then infiltrated with Spurr’s resin, flat embedded and sectioned on a Dupont Sorvall MT2-B ultramicrotome. Specimens where viewed with a Philips EM 201 transmission electron microscope at accelerating voltages of 60 and 80 kV. 111 Ascospore measurements were made in water at x1000 magnification. 30 specimens were examined. For each specimen width and length of 20 spores was measured. The values given in parentheses represent the smallest and the largest measurements respectively. The range of the mean values calculated for each specimen lies between these extremes and is given as the lowest mean value and the highest mean value. An attempt was made to evaluate how many spores did have a conspicuous microrugulate ornamentation and / or a median wall thickening. For 40 spores of 25 specimens the amount of spores with or without ornamentation and with or without thickening was recorded. The average of these records is given. The width of the conidium was less than 1 µm and thus smaller than the scale ocular micrometer at a x1000 magnification. Therefore only conidium length was recorded. Pycnidia in Buellia dispersa are rare and only 40 conidia of five specimens could be measured. RESULTS BUELLIA DISPERSA A. Massal. Schedul. Critic. 8:150. 1856. TYPE: Nel fossato di Granarolo, 22 January 1853, Baglietto s.n. (VER–lectotype!, selected by Scheidegger 1993) B. italica var. tumida A. Massal., Sched. Critic. 9: 163. 1856.—B. tumida Bagl., Mem. Reale Acad. Sci. Torino, Ser. 2, 17:423. 1857. TYPE: ITALY. Ad saxa micaceo-schistosa Liguria propre oppidum Voltri (Bosco dell’ aqua Santa), Baglietto s.n. [= Lich. Exs. Ital. no. 303] (TO–isotype!, holotype not seen). 112 Lecidea squamulata A. Nyl., Flora, Jena 56: 201. 1873; Bull. Soc. Linn. Normandie, Sér. 2, 6: 311. 1873. TYPE: FRANCE. PYRÉNÉES ORIENTALES. Collioure. 4 July 1872, Nylander s.n. (H-NYL 9229b–holotype!). Lecidea dispersa var. subeffigurans A. Nyl. in Lamy, Bull. Soc. Fr. 30: 421. 1883. TYPE: FRANCE. Sur du schiste à Lourdes, Lamy s.n. (H-NYL 9228–isotype!, holotype not seen). Buellia retrovertens Tuck., Syn. N. Amer. Lich. 2: 89. 1888. TYPE: U.S.A. Granitic rocks, Rocky Mountains in Colorado, 1879, Brandegee s.n. in Herb. Sprague [ex Tuckerman sheet no 3281] (FH–isotype!, holotype not seen). 113 TABLE 3. Thallus characteristics of morphotypes of Buellia dispersa. thallus growth areoles central marginal thallus color pruina morphotype I distinctly areolate areoles inflated to subsquamulose ± dispersed growing in irregular patches beige to brown or gray to olive epruinose to finely pruinose morphotype II areolate, typically forming rosettes contorted and strongly fissured beige to brown, often concealed by pruina characteristic thick and coarse pruina morphotype III areolate, forming distinct rosettes areoles flattened angular sublobate ivory to beige-brown or pale gray to lead gray usually with fine pruina frequently forming distinct lobes 114 → FIGURE 14. Typical morphotype (morphotype I) of Buellia dispersa.—A. Bullate, beige thallus with pruina; isotype of B. retrovertens (Brandegee s.n. in Herb. Sprague [ex Tuckerman sheet no 3281], FH).—B. Subsquamulose, ± contiguous, beige thallus with little pruina (Nash 9605, ASU).—C. Subsquamulose, ± dispersed, beige thallus with pruina; isotype of B. tucsonensis (Blumer 5795, MICH).—D. Subsquamulose, dispersed, beige thallus with pruina, very small areoles; holotype of B. blumeri (Blumer 5750, W).— E. Subsquamulose, contiguous, dark olive grey thallus with almost no pruina (Nash 40069, ASU). 115 116 → FIGURE 15. Morphotype II and morphotype III.—A. Morphotype II: Distinctly fissured thallus with coarse pruina, lobate marginal areoles and contorted, central areoles (Nash 8951, ASU).—B. Morphotype II: Thallus lacking marginal lobes, areoles distinctly fissured and with coarse pruina (Darrow 5138, ASU).—C. Morphotype III: Pruinose apothecium on a central subsquamulose areole (Nash 40568, ASU).—D. Scanning electron micrograph from the same specimen (Nash 40568, ASU).—E. Morphotype III: with lobed marginal areole not fissured and finely pruinose [Isotype of Buellia tergestina, Schuler s.n. (Krypt. Exs. Vindob. no. 58), S]. 117 118 → FIGURE 16. Light micrographs of thallus, ascospores and ascus (DIC).—A. Section through the thallus with epinecral layer, cortical layer, algal layer and medulla.—B. Squash preparation of a juvenile ascus.—C. Squash preparation of an ascus shortly before maturity, the spores have a conspicuous median thickening.—D. Microrugulate ascospore. 119 120 → FIGURE 17. Light, transmission electron and scanning electron micrographs of B. dispersa.—A. Cross section through the dispersa-type exciple (light micrograph, DIC).— B. Transmission electron micrograph of an ascospore: (1) perispore, (2) intermediate layer, (3) proper wall, (4) endospore, (M) mitochondrion, (N) nuclei, (P) septal pore, (V) vacuole, arrows indicate concentric bodies, the asterix indicates a complex membrane structure often found in close proximity to the septal pore.—C. Section through the apothecium and thallus (scanning electron micrograph, SEM): (a) algal layer, (c) phenocortex, (e) exciple, (h) hymenium, (m) medulla.—D. Ascospores (SEM): the spores were cut during a developmental stage when no ornamentation (ps) is visible and the spores septum (s) is not thickened.—E. Section through the exciple (SEM).—G. Inner exciple (SEM): most hyphae have been cut longitudinally.—G. Outer exciple (SEM): most hyphae have been cut in cross section. 121 122 Buellia tergestina J. Steiner & Zahlbr. in Zahlbr., Annal. Naturhist. Hofmus. Wien 9: 134. 1894. TYPE: ITALY. Litorale austriacum, ad saxa arenaria in agro tergestina, Schuler s.n. [Krypt. Exs. Vindob. no. 58] (GZU, S, US, W–isotypes!, holotype not seen). Buellia blumeri Zahlbr., Annal. Mycol. 7: 477. 1909. TYPE: U.S.A. ARIZONA. On Basalt, Tucson, March 1908, Blumer 6777 [= Herb. Mus. Palat Vindob. Acqu. 1909 no. 5750] (W–holotype!; MICH–isotype!). Buellia tucsonensis Zahlbr., Annal. Mycol. 7: 477. 1909. TYPE: U.S.A. ARIZONA. On basalt, Tucson, March 1908, Blumer 6795 (MICH–isotype!, holotype not seen). Buellia duartei Samp., Liquen. Ined.: 1. 1920. TYPE: Portugal, Povao de Lanhoso, 29 September 1919, Sampaio s.n. (W–isotype!, holotype not seen). Buellia cinereoglauca de Lesd., Annal. Crypt. Exot. 5: 127. 1932. TYPE: U.S.A. NEW MEXICO. Lagunitas, Soldier’s camp, sur roches siliceuses; holotype not designated in the protologue, in need of lectotypification. Specimen examined: Lagunitas, Soldier’s camp, sur roches siliceuses, March 1927, Arsène 19727 (US!). Taxonomic Note.—Imshaug (1951) mentions B. puebla de Lesd. and B. puebla var. plana de Lesd. as synonyms. Scheidegger (1993) also mentions Buellia subsquamosa sensu Buschardt 1979, non J. Steiner 1907. Type material of these taxa was, however, not available for our study. Thallus crustose; distinctly areolate; areoles widely dispersed in irregular patches or closely aggregated and typically forming rosettes with marginal lobes; surface smooth to deeply fissured with fine or coarse pruina, rarely epruinose; thallus color ivory, beige to deep brown, gray to rarely dark olive, often concealed by abundant pruina. Three 123 morphotypes can be distinguished (Table 3). Apothecia sessile, black lecideine, plane to convex; exciple of dispersa-type, i.e. hyphae radially arranged, outer exciple textura angularis, brown, HNO3-; inner exciple textura oblita, continuous with the dark reddish brown hypothecium; paraphyses simple to moderately branched, apically swollen, with brown pigmented cap. Asci 8-spored, Bacidia-type; Ascospores narrowly-oblong to ellipsoidal, (6)7–8(8.5) x (10)12–14(18) µm, proper septum narrow but briefly with Physconia-type median thickening, with septal pore channel, simple pore and undifferentiated pore plug; ornamentation inconspicuous, microrugulate at maturity; spore wall differentiated into perispore, intermediary layer, proper wall and endospore. Conidiomata flask-shaped, simple pycnidia; mature pycnidia globose, almost entirely occupied by densely branched conidiophores; conidiogenous cells intercalary and terminal (cf. conidiophore-type V sensu VOBIS 1980); pycnidial development similar to the Umbilicaria-type sensu Vobis (1980) and Vobis & Hawksworth (1981); conidia simple, bacilliform, 5-6 µm long. Chemistry:.—Major compound atranorin, typically accompanied by 2'-Omethylperlatolic and ± confluentic acid; rarely an unknown substance (pale green after charring, UV+ dark, Rf-class 6/6/6 in A, B’, C; found in the following specimens: Moberg 8902, 10421; Nash 26092, 29579; Sipman 25026, 25125; Scheidegger field no. 123, 130, 160, 164, 165, 260). Thallus typically K+ yellow, P+ yellow, C-, KC-, CK(reactions can be weak according to the concentration). Medulla I- (important: always test with the compound microscope, positive reactions with Lugol’s iodine can be very weak!). 124 DISCUSSION Thallus Color, Morphology and Pruinosity.—Thallus morphology of Buellia dispersa is extremely variable and several morphotypes can be distinguished (Table 3): Morphotype I: This typical morphotype has a distinctly areolate thallus (Fig. 14). The thalli do not form rosettes and the thallus outline is rather irregular. Individual areoles can be dispersed and are then often inflated (= bullate, tumid, Fig. 14A) or become subsquamulose along the areole margins (Fig. 14C). Other thalli have more closely aggregated areoles, the single areoles having an angular outline (Fig. 14B,E). Areoles have a smooth surface which may be obscured by pruina. Pruinosity and color vary. Heavily pruinose specimens appear whitish to ivory, less pruinose specimens are either beige to deep brown or a pale gray to deep lead or olive gray. Morphotype II: This morphotype is very distinct (Fig. 15A,B). It is composed of closely aggregated areoles, typically forming rosettes delimited from beige marginal lobes. Although marginal lobes are usually very conspicuous (Fig. 15A), they are not present in all specimens of this morphotype. The central areoles are often much contorted and have a cracked or fissured surface. This may give the entire thallus a somewhat rimose appearance (Fig. 15B). However, the central areoles are nevertheless clearly delimited. They are densely covered with a coarse, white pruina whereas marginal lobes show little pruinosity and have a smooth, beige surface. This morphotype has not yet been observed among the European material. In the Sonoran Desert Region most specimens have been collected at elevations above 1000 m. However, some have also been found at elevations as low as 300 m. 125 Morphotype III: The third morphotype usually forms rosettes (Fig. 15C-E). Like the first morphotype it is distinctly areolate, but the entire thallus is more contiguous and only the marginal areoles become dispersed. Marginal areoles are frequently sublobate whereas closely adjoining central areoles are angular. European specimens of this morphotype have been called B. tergestina or B. tumida (Fig. 15E, also see Scheidegger 1993). The North American material has a fine pruinose appearance (Fig. 15C) and marginal areoles are often more or less elongated. European specimens show little pruinosity and have stouter marginal areoles (Fig. 15E). All material has a smooth surface structure and the thallus color varies from pale ivory, beige to pale gray or lead gray. The great variation of the different morphotypes may be explained by differences in substrate composition, erosion pressure and light availability. The cortical layer of B. dispersa is differentiated into an amorphous layer of dead organic material and a layer below with densely interwoven hyphae (Fig. 16A). The epinecral material may disintegrate exposing the cortical hyphae below. In most specimens this layer is densely packed with fine crystals. Some of these crystals dissolve in KOH, others do not dissolve in KOH and concentrated HNO3. The composition of the crystals is currently being investigated using x-ray diffraction. Scheidegger (1993) argued that substrate composition, especially CaCO3, accounts for the variation of crystal content and thus pruinosity of the specimens. However, most specimens grow on rock substrates which do not react with HCl. Therefore, we assume that other minerals rather than CaCO3 play a major role in the accumulation of crystals in 126 the cortex. If the epinecral layer remains intact the crystals inside the cortical layer are not exposed. The thallus surface then appears smooth and devoid of pruina. Consequently, we suggest that the variation in thallus pruinosity can be ascribed to differences in erosion pressure as well as substrate variation. The cortical layer below the epinecral organic material is characterized by a beige to pale brown pigmentation. This pigmentation is probably identical with the brown apothecial pigment and seems to be confined to the cell walls of the hyphal tips. In KOH most of this thallus pigmentation dissolves. However, within the thallus pigments are much less concentrated than in the apothecium, and a KOH+ yellow solution characteristic for the soluble apothecial pigment was not observed even in thick thallus sections. Although it is variable in amount, the brown pigmentation usually shines through the thallus surface and many specimens therefore have a pale beige to brown color. With chlor-zinc-iodine dead algal cells stain weakly violet within the cortical layer. Living algal cells below intergrade into this layer and the cortex is thus a phenocortex (“Scheinrinde” sensu Poelt 1958). The algal layer may be rather thin and composed of loosely packed algal cells or it forms a thick matt of densely packed cells, which probably accounts for the gray to dark olive gray color of some of the morphotypes. Specimens with dark thalli and thicker algal layer are presumed to be more abundant in shaded microhabitats. Variation in growth morphology is difficult to explain. We have observed that areoles of morphotype I tend to grow more closely together along crevices and rock fissures. On 127 irregular surfaces these areoles become contorted and frequently sublobate. In contrast, distinct rosettes seem to result from unobstructed circular growth on smooth substrates. Areoles of morphotype III are generally more widely dispersed than areoles of morphotype II which are typically closely adjoined and contorted even on fine-grained sandstone. In both morphotypes marginal lobes develop best where they are not hindered by an irregular surface or by competition with other, closely adjoining thalli. Intergradation of the various thallus morphologies occurs and the morphotypes are less distinct than the descriptions suggest. We did not find any correlation of the morphology with any other anatomical or chemical characters of the group. Until molecular data allows us to evaluate the variation more accurately, it does not seem appropriate to assign formal rank to these morphotypes. The immense thallus variation in Buellia dispersa, however, raises the question to which extent thallus lobation is subject to environmental modification. The crustose lichen genus Dimelaena is commonly distinguished from Rinodina and Buellia by a distinct lobate margin. Rico et al. (2000) suggested that Dimelaena radiata and Buellia fimbriata are two very closely related taxa which have unfortunately been assigned to two different genera of crustose Physciaceae. If differences of thallus lobation can occur within one species, it would be interesting to investigate the phenomenon more closely. We therefore suggest to re-evaluate the segregation of Dimelaena on the basis of thallus lobation. Apothecial Morphology and Anatomy.—The hymenium of B. dispersa is not interspersed with oil droplets. Pigmentation of the outer exciple extends across the 128 hymenium as a dull brown epihymenium. This pigmentation appears in (or on) the upper part of the hyphal cell wall of the capitate end cells of the paraphyses. It thus looks like a strongly pigmented cap on top of the little branched paraphyses. The asci belong to the Bacidia-type sensu Rambold et al. (1994) (Fig. 16B,C). Only at very young stages are apothecia immersed in the thallus areoles. Apothecia soon become sessile and are typically lecideine. The margin only very rarely appears dark brown rather than black. Mature apothecia are characterized by a plane disk and a conspicuous margin. Old apothecia may become convex but the margin is very rarely excluded. The apothecia are usually shiny black (Figs. 14 and 15A,B, E) and very rarely have an inconspicuous fine pruina (Fig. 15C). By distinguishing several exciple types Scheidegger (1993) questioned the traditional segregation of Buellia with lecideine versus Rinodina with lecanorine apothecia. The distinction is not as clear-cut as the original terminology suggests. While the dispersatype is distinctly lecideine, other exciple types show more variation. The internal arrangement of hyphae within the fungal plectenchyma is difficult to observe and only reluctantly have these characters been recognized as taxonomically important. The dispersa-type shows little variation and a more detailed description may help to recognize the distinct anatomy. Hyphae of the dispersa-type exciple are ± radially arranged, relatively short celled and closely aggregated. They can be distinguished in thin sections (< 14 µm) with conventional or phase contrast light microscopy. However, plectenchyma structure appears much more plastic in differential interference contrast (DIC, Fig. 17A) where 129 separate hyphae can be distinguished more clearly. The inner exciple is continuous with the hypothecium. It has a dark reddish brown color and extends far into the thallus. The hyphae of the inner exciple can be described as textura oblita, i.e. they radiate more or less parallel from the hypothecium into the inner exciple and characteristically have narrow lumina and thickened cell walls (Korf 1973). Below the hymenium these hyphae grow rather irregularly and a gradual transition from textura oblita to a densely interwoven textura intricata can be observed. Although the outer exciple is rather similar to the inner exciple, the cells here become gradually shorter and more rounded. Consequently Scheidegger (1993) distinguishes the outer hyphae as textura angularis, i.e. composed of polyhedral cells without inter-hyphal spaces (Korf 1973). In the light microscope these hyphae appear to have thinner cell walls than the parallel hyphae of the inner exciple. The cells of the outer exciple are characteristically dull brown and there is commonly a pale transition zone between these outer cells and the parallel, dark red-brown hyphae of the inner exciple. In older apothecia the pigmentation gradually extents across the transition zone and the exciple may become strongly brown throughout. Within the exciple Scheidegger (1993) distinguished two brown pigments, one insoluble in KOH and one soluble as a faint yellow solution. This reaction is usually faint and can be best observed in thick sections. Both pigments appear to be confined to the cell walls. An aeruginose HNO3+ wine-red pigment characteristic for the aethalea-type has not been observed in any of the specimens. 130 Our SEM studies demonstrate that a distinction of fungal plectenchyma types is not as clear-cut as some of the light microscopic observations suggest (Fig. 17E). A gradual change in the orientation and shape of the hyphal cells can be observed and the cell walls appear ± evenly thickened throughout. In the inner exciple the fungal hyphae run ± parallel to the section (Fig. 17F). Thus, they have mostly been cut longitudinally. Towards the outer exciple, cells become shorter and are mostly cut in cross section (Fig. 17G). There is a transitional zone between the inner and outer exciple where the section apparently cuts tangentially through the hyphal cell. The polyhedral shape of the cells in the outer exciple is therefore partially the result of a change in hyphal orientation, and it does not necessarily represent a distinctly separate plectenchyma type. Whether these outer cells are truly more isodiametric is difficult to decide, even with the SEM. A gradual transition from a pseudoparenchymatous outer exciple towards a more densely packed prosoplectenchymatous inner exciple as well as the absence of the aeruginose, HNO3+ wine-red pigment is regarded as characteristic for the dispersa-type sensu Scheidegger (1993). Ascospore Ontogeny, Spore Typification and Ornamentation.—One of the characters that segregate Buellia from Rinodina and Dimelaena has traditionally been the Buelliaspore type with a simple septum that does not become thickened during spore ontogeny. Scheidegger (1993), however, realized that in Buellia dispersa a median thickening can frequently be observed. Accordingly, he reclassified the spores as Physconia-type ascospores. This type was originally introduced by Poelt (1956) to describe large spores with a median thickening and a conspicuously warty surface. Mayrhofer & Poelt (1979) 131 mention that Rinodina sicula has a spore type similar to the Physconia-type but with a less conspicuous fine-warty spore surface. Because of the presence of a torus, Mayrhofer (1982; Mayrhofer 1984a, b) later designated a separate spore type as the Sicula-type. In contrast the Physconia-type has no conspicuous torus. Scheidegger (1993, p. 333), however, considers the torus as “highly dependent on the development of the pigmentation of the spore wall”. He therefore does not distinguish between spore types with or without a torus. Nordin (1997, p. 205) emphasizes that there is “no support for the distinction of a torus as a structurally different part in the spore wall.” In the specimens of Buellia dispersa which we have examined no torus could be observed. Characters like septum formation, spore wall pigmentation (torus) and spore ornamentation do not necessarily correlate and it is therefore important to notice that “spore types” in Physciaceae often represent only stages of the spore ontogeny. These stages are not necessarily present during the entire development of the ascospore. In B. dispersa septa form very early during spore formation and non-septate spores are rarely observed. However, the stage when the spore septum forms a distinct median thickening represents only a very brief phase shortly before ascospore maturity (Fig. 16C). During this stage spores are usually less strongly pigmented and often appear gray-green instead of dark brown. The median thickening soon disappears in mature spores. In squash preparations of 25 randomly selected apothecia from all different morphotypes, thinseptate spores were nine times more abundant (n = 1000) than spores with median thickenings. In some specimens spores with a typical Physconia-type septum were only found if several apothecia were examined, and even then only a few spores with median 132 thickening were found. The differences may correlate with the time when the specimen was collected. Because the ontogenetical phase with a distinct median thickening along the septum is very brief, specimens collected during active spore production are likely to have more spores at this stage. Similar observations were made when spore ornamentation was studied. Microrugulate ascospore ornamentation is usually distinct only during the last stages of spore ontogeny. In light microscopy it is most conspicuous with DIC and only visible at 1000x magnification with apertures of 1.3 or higher (Fig. 16D). Immature spores with no ornamentation are usually much more abundant and will therefore be over-represented in both TEM and SEM studies (Fig. 17B,D). In squash preparations of 25 randomly selected apothecia from all different morphotypes, spores without ornamentation were to times more abundant (n = 1000) than spores with ornamentation. The microrugulate ornamentation can thus easily be overlooked even if spores are studied with SEM or TEM. Scheidegger (1993) emphasizes the necessity of studying spores with the light microscope because SEM studies may be hindered by a proteinaceous mucilage on the spore surface. A simple incubation in commercially available washing powder (Honegger 1985) removes this mucilage. However, further studies are necessary to examine the effect of this treatment on ascospore ornamentation. The origin of the microrugulate ornamentation is not evident from our studies. Ascospores which were examined with TEM have a rather thin wall differentiated into a perispore, intermediary layer, a proper wall and an endospore (Fig. 17B). A distinct 133 ornamentation of the perispore could not be observed. We assume that the microrugulate ornamentation develops at a later stage than the spores examined. The spore is clearly euseptate, i.e. both proper wall and endospore form a septum along a central septal plate (see Nordin 1997 for a discussion). The septum is perforated by a central pore that is blocked by an undifferentiated pore plug. Infiltration of the spore cytoplasm with fixatives and resin is hindered by the thick spore wall and cell organelles were rarely well preserved. However, in some preparations mitochondria, nuclei and concentric bodies could be distinguished in both cells of the ascospores. ACKNOWLEDGEMENTS We like to thank the curators of the herbaria cited for the loan of the material and are much indebted to Bill Sharp and Dr. Robert Roberson (Arizona State University, ASU) and Dr. Rosemarie Honegger (University of Zürich) for their valuable advice on scanning and transmission electron microscopy. Martin Frei (WSL Birmensdorf) carried out the thin layer chromatography of the American specimens from the private herbarium of Dr. Christoph Scheidegger. Dr. Bruce Ryan, Dr. Don Pinkava and Robin Schoeninger (ASU) gave valuable advice on taxonomy and typification. Dr. Corinna Gries (ASU) helped with generating the distribution maps. Dennis McDaniel printed out the final version of the photographic plates. This research was supported by a grant from the National Science Foundation for the Sonoran Desert Lichen Flora Project (Grant No. DEB 97-06984 and BSR 96-29532). 134 LITERATURE CITED CULBERSON, C. F. & H. KRISTINSSON. 1970. A standardized method for the identification of lichen products. Journal of Chromatography 46: 85-93. CULBERSON, C. F. & A. JOHNSON. 1982. Substitution of methyl tert.-butyl ether for diethyl ether in standardized thin-layer chromatographic method for lichen products. Journal of Chromatography 238: 438-487. HONEGGER, R. 1985. Scanning electron microscopy of the fungus-plant cell interface: A simple preparative technique. Transactions of the British Mycological Society 84: 350-533. IMSHAUG, H. A. 1951. The Lichen-forming Species of the Genus Buellia in the United States and Canada. University Microfilms. Ann Arbor, Michigan. Publication No. 2607. KORF, R. P. 1973. Japanese discomycete notes I-VIII, pp. In AINSWORTH, G. C., F. K. SPARROW & A. S. SUSSMAN (eds.), The Fungi; IVa. A taxonomic treatment with keys. Ascomycetes and fungi imperfecti. Academic Press, London. MAYRHOFER, H. 1982. Ascosporen und Evolution der Flechtenfamilie Physciaceae. Journal of the Hattori Botanical Laboratory 52: 313-321. ———. 1984a. The saxicolous species of Dimelaena, Rinodina and Rinodinella in Australia, pp. 511-536 Beitrage zur Lichenologie. Festschrift J. Poelt. Beiheft zur Nova Hedwigia 79. J. Cramer, Vaduz. ———. 1984b. Die saxicolen Arten der Flechtengattungen Rinodina und Rinodinella in der Alten Welt. Journal of the Hattori Botanical Laboratory 55: 327-493. 135 MAYRHOFER, H. & J. POELT. 1979. Die saxicolen Arten der Flechtengattung Rinodina in Europa. Bibliotheca Lichenologica 12: 1-186. MIETZSCH, E., H. T. LUMBSCH & J. A. ELIX. 1994. WINTABOLITES (Mactabolites for Windows). Users manual and computer program. University Essen, Essen. NORDIN, A. 1997. Ascospore characters in Physciaceae: an ultrastructural study, pp. 195208. In TIBELL, L. & I. HEDBERG (eds.), Lichen Studies Dedicated to Rolf Santesson. Symbolae Botanicae Upsalienses, Acta Universitatis Upsaliensis, Uppsala. POELT, J. 1956. Zur Systematik der Flechtenfamilie Physciaceae. Nova Hedwigia 9: 2123. ———. 1958. Die lobaten Arten der Flechtengattung Lecanora Ach. sensu ampl. in der Holarktis. Mitteilungen der botanischen Staatssammlung München 20: 411-589. RAMBOLD, G., H. MAYRHOFER & M. MATZER. 1994. On the ascus types in the Physciaceae (Lecanorales). Plant Systematics and Evolution 192: 31-40. RICO, V. J., V. CALATAYUD & M. GIRALT. 2000. The lichen genus Dimelaena in the Iberian Peninsula, pp. 25 The Fourth IAL Symposium, Progress and Problems in Lichenology at the Turn of the Millennium. Universitat de Barcelona, Barcelona. SCHEIDEGGER, C. 1993. A revision of European saxicolous species of the genus Buellia De Not. and formerly included genera. Lichenologist 25: 315-364. VOBIS, G. 1980. Bau und Entwicklung der Flechten-Pycnidien und ihrer Conidien. Bibliotheca Lichenologica 14: 1-141. 136 VOBIS, G. & D. L. HAWKSWORTH. 1981. Conidial lichen-forming fungi, pp. 245-273. In COLE, G. T. & B. KENDRICK (eds.), 1: Biology of Conidial Fungi. Academic Press, New York. WHITE, F. J. & P. W. JAMES. 1985. New guide to microchemical techniques for the identification of lichen substances. British Lichen Society Bulletin 57: 1-41. 137 PUBLICATION 2 (The Bryologist 107, 2004): Buellia turgescens is synonymous with Buellia badia and must not be included in Amandinea FRANK BUNGARTZ and THOMAS H. NASH III Arizona State University Lichen Herbarium, School of Life Sciences, P. O. Box 87 4501, Tempe, AZ 85287 – 4501, U.S.A. e-mail: frank.bungartz@asu.edu; tom.nash@asu.edu Abstract. A comparison of Buellia turgescens Nyl. ex Tuck. with B. badia (Fr.) A. Massal. demonstrates that the two taxa are identical. Both have very similar thalli, identical apothecial anatomy, identical size and structure of the ascospores, and bacilliform conidia. Consequently, B. turgescens should not be included in the genus Amandinea, which was erected to accommodate species of Buellia s.l. and Rinodina s.l. with filiform conidia. Thalli of B. badia show juvenile parasitism on foliose and crustose lichens, but with age become independent. The distribution of B. badia is shown in the southwestern North America. During our studies of the saxicolous Buellia species with one-septate ascospores from the Sonoran Desert Region we compared type material of both Buellia turgescens Nyl. ex Tuck. and B. badia (Fr.) A. Massal. Imshaug (1951) first noticed that these two taxa are very similar. Together with Buellia punctata (Hoffm.) A. Massal., he included them in 138 his "stirps punctata". Scheidegger (1993) resurrected the genus Amandinea and included B. punctata because of filiform conidia. Marbach (2000) recently suggested transfering B. turgescens into Amandinea even though he did not find conidia among the material examined. He argued that the genus concept was emended by Sheard & May (1997), when those authors transferred several North American species of Buellia s. l. with filiform conidia into Amandinea. We do not agree with Marbach's (2000) assessment. All species included in Amandinea by Sheard & May (1997) have filiform conidia and the authors do not propose additional characters to justify transferring species into Amandinea (Sheard, pers. comm.). The inclusion of a species within Amandinea is questionable if conidia have not been observed, and a transfer of species with bacilliform conidia can certainly not be justified. In the case of B. turgescens we demonstrate the occurrence of bacilliform conidia, and its transfer into Amandinea can, therefore, not be accepted. Furthermore, we provide evidence that Buellia turgescens is identical with Buellia badia and must be regarded as a synonym of that species. METHODS All specimens were examined with light microscopy using hand- and cryo-sections. Both conventional bright field microscopy (BF) as well as differential interference contrast (DIC) was used. All specimens were spot tested and routinely examined with standardized thin layer chromatography (Culberson & Kristinsson 1970; Culberson & Johnson 1982; White & James 1985; Orange et al. 2001). TLC-Plates were interpreted with the computer program WINTABOLITES (Mietzsch et al. 1994), and scanned for 139 permanent record (Egan 2001). Spores measurements are given according to Nordin 2000). Pigment names follow Meyer & Printzen (2000). RESULTS AND DISCUSSION BUELLIA BADIA (Fr.) A. Massal., Memor. Lichenogr. 1853: 124. 1853. Lecidea badia Fr., Syst. Orb. Veget. 1: 287. 1825. TYPE: AUSTRIA. KÄRNTEN. On the Southern Slope of the Falkenberg near Klagenfurt [original label data: am Südabhang des Falkenbergs bei Klagenfurt”], 1890, J. Steiner (Arnold, Lich. Exs. no. 1505, UPS– neotype!, selected by Foucard, Moberg, & Nordin, Nordic Lichen Flora 2: 70. 2002). Buellia turgescens Nyl. ex Tuck., Gen. Lich.: An arrangement of the North American Lichens: 185. 1872. Lecidea turgescens Nyl., Mém. Soc. Sci. Nat. Cherb. 5: 337. 1857. nomen nudum.—Amandinea turgescens (Tuck.) Marbach, Bibl. Lich. 74: 109. 2000. comb. inval. TYPE: U.S.A. MASSACHUSETTS [Nova Anglia]: ex herb. Tuckerman 5538 (post mortem Nylanderi insertum, H-NYL 9534–lectotype!, selected by Marbach, Bibl. Lich. 74: 109. 2000). Note: Most of the label data on the lectotype specimen are illegible. Neither Nylander (1858 ('1857')) nor Tuckerman (1872) give any specific information where exactly the type specimen was originally collected. Buellia coniopotiza (Nyl.) de Lesd., Bull Soc. Bot. France 53: 685. 1907.—Lecidea conioptiza Nyl. Flora 61: 244. 1878. TYPE: FRANCE. HAUTE VIENNE. On schist near Châlucet, very rare species [original label data: Sur un rocher schisteux près de Châlucet, espèce très rare], 28 July 1877, Lamy s.n. (H-NYL 9537a–holotype, M-002374–isotype!) 140 Buellia turgescentoides Fink, Ohio Biol. Surv. 2: 349. 1921. Type: U.S.A. OHIO. Lake County: rock, open field, near Painesville [41°45'N, 81°14'W]. 15 October 1915, B. Fink 739 (MICH–holotype!). For additional synonyms see Scheidegger (1993) and Hafellner (1979). Thallus (Fig. 18) crustose, bullate, subsquamulose to distinctly squamulose; without prothallus; surface matt to shiny, deep “chocolate” to grayish brown, epruinose; phenocorticate with thin epinecral layer of dead cells; no crystals within medulla (Fig. 19G). Apothecia soon sessile on thallus surface; lecideine, 0.3–0.8(–0.9) mm in diameter; proper margin thin, becoming excluded with age; disk and margin black, epruinose, plane to convex; exciple narrow (aethalea-type sensu Scheidegger 1993, Fig. 19A,E,H,K), poorly differentiated, inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), similar in structure and orientation to paraphyses, often ± reduced and transient with dull reddish brown hypothecium (leptoclinoides-brown, textura intricata); outer excipular hyphae parallel (textura oblita), cells swollen and usually strongly carbonized with various amounts of brown pigment (elachista-brown), pigmentation continuous with epihymenium; paraphyses not inspersed, simple to moderately branched, apically swollen, with brown pigment cap (elachista-brown). Asci 8-spored, clavate, Bacidia-type (Fig. 19B,F); Ascospores (Fig. 18B,F,I,L) oblong to ellipsoid, not constricted, with obtuse ends, not curved, (10.0–)10.5–[11.9]–13.2(–15.0) x (5.0–)5.8[6.5]–7.3(–8.0) µm (n = 200) (measurement format according to Nordin 2000), young spores olive brown, mature and old spores deep brown, one-septate, septum narrow, not 141 thickening during spore ontogeny, old spores indistinctly microrugulate. Pycnidia rare, urceolate to globose, unilocular (Fig. 19C); pycnidium walls lined with short, barely branched conidiophores, conidiogenous cells terminal (conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to Umbilicaria-type sensu Vobis (1980) and Vobis & Hawksworth (1981); conidia (Fig. 19C,D,J,M) simple, bacilliform, 3.0–5.0 µm x 1.0–1.5 µm (n = 50). Chemistry.—No substances found in independent thalli; parasitic thalli are difficult to isolate and may contain substances from the host lichen. Thallus spot test reactions negative, cortex and medulla UV-. Medulla I-. Substrate and ecology.—Initially parasitic on a variety of crustose and foliose lichen genera, eventually establishing independent thalli over rock or decorticated wood, rarely on bark. Distribution (Fig. 7).—Widespread but not common. Known from Europe, Northern Africa and North America. The distribution in the Sonoran Desert Region is shown in Fig. 2.3. Additional specimens examined.—CANADA. BRITISH COLUMBIA. Ryan 31628a (ASU). CZECH REPUBLIC: MORAVIA. Vĕzda s.n. (MSC-67474). GERMANY. Körber s.n. (L-0065266, L-0065265, L-0065267). ITALY. LIGURIA. Poelt s.n. (M-0061336. Note: this specimen grows on Dimelaena oreina together with B. imshaugii, another parasitic species of Buellia); Gbarbano s.n. (MSC-146450). PIEDMONT. Gbarbano s.n. (MSC146450). MEXICO. BAJA CALIFORNIA. Scheidegger s.n. (hb. Scheidegger). CHIHIAHUA. Weber & Bye L-65221 (COLO). SONORA. Wetmore 69862 (MIN). U.S.A. ARIZONA. 142 Apache Co., Nash 26814b; Cochise Co., Nash 3713c; Maricopa Co., Nash 18444; Pinal Co., Nash 6154; Santa Cruz Co., Nash 25244 (ASU); Coconino Co., Nordin 5253 (UPS). CALIFORNIA. Los Angeles Co., Hasse Lich. Exs. 179, Knudsen 348b (ASU); Hasse s. n. (FH); Monterey Co., Bratt 5607; Napa Co., Tucker 6299 (ASU); Riverside Co., Weber 82059 (COLO), ex Her. Hasse 3026 (FH); San Diego Co., Bratt 4313 (SBBG); Santa Clara Co., Herre 1101 (FH); Solano Co., Weber L-42097 (COLO); Tulare Co., Ryan 26667; Ventura Co., Nash 37098 (ASU). COLORADO. Boulder Co., Imshaug 19057 (MSC-48036). CONNECTICUT. Fairfield Co., Evans 790 (FH). IDAHO. Twin Falls Co., Ryan 32914 (ASU). IOWA. Fayette Co., Fink 1893 (MSC-50937). MASSACHUSETTS. Barnstable Co., Brodo 4415 (MSC-64583); Ravenel s.n. (BM-000660155); Tuckerman 53.365 (ex Tuckerman Sheet 3249; FH) [Note: annotated by Imshaug in 1951 as: “Buellia punctata (Hoffm.) Mass. 53.365 = holotype of Buellia turgescens Tuck”]; Sprague s.n. (W, Vindob. acq. 1927 no. 12082) [Note: This specimen has erroneously been labeled as a type. The material was collected by Sprague, who annotated it as “sp. orig. videtae” meaning “original material seen”. The specimen is clearly not part of the type.]. MINNESOTA. Yellow Medicine Co., Fink s. n. (FH). OHIO. Hocking Co., Ryan 21896 (ASU); Lake Co., Fink 739 (FH). NEW YORK. Suffolk Co., Brodo 2371 (MSC-58302). OREGON. Lake Co., Ryan 28471b (ASU). UTAH. Grand Co., Ryan 11660 (ASU). Taxonomy.—The name B. turgescens has caused considerable taxonomic confusion. It first appears in Nylander (1858 ('1857')). All Nylander stated about this taxon is, “... Lecidea turgescens Nyl., in Nova Anglia ...” (Nylander 1858 ('1857'), p. 335). According to the Botanical Code (Greuter et al. 2000, Article 32.1c), a validly published name must 143 be accompanied by a diagnosis sensu article 32.2: “A diagnosis of a taxon is a statement of that which in the opinion of its author distinguishes the taxon from others.” The fact that Nylander mentioned Lecidea turgescens as a species from New England, does not indicate how this taxon is distinguished from any other species in that area. His statement therefore cannot be accepted as a diagnosis. The name L. turgescens is thus a nomen nudum and not the basionym of B. turgescens. Tuckerman (1872) validated the name by providing the first effective publication with a scant but valid species description: “B. badia ... The lichen is comparable also with the (exclusively lignicoline) B. turgescens (Nyl.) of New England, scarcely indeed differing at all in the spores; and B. turgescens, it is further observable, resembles still more closely Lecidea insularis, Nyl., which Flotow was inclined (Koerb. Syst. p. 239) – without, it is evident, microscopical investigation – to regard as a variety of B. badia.” Tuckerman 1872, p. 185). His diagnosis “... scarcely indeed differing at all in the spores ...” is written in English, not in Latin, but published before 1 January 1935, and therefore valid according to article 36.1. of the Code. Because the description was published before 1 January 1958, the name is also valid, even though no type was designated by Tuckerman (article 37.1.). The basionym of the species should therefore be cited as Buellia turgescens Nyl. ex Tuck. When Marbach (2000) transferred B. turgescens into Amandinea, he designated a specimen in UPS as the lectotype [ex herbarium Tuckerman 5538, post mortem Nylanderi insertum (9534-H-Nyl.), Figs. 18E,F, 19H-I]. In 1951 Imshaug annotated a different specimen as the type as follows: “Buellia punctata (Hoffm.) Mass. 53.365 = holotype of Buellia turgescens Tuck.” [no. 53.365 (ex Tuckerman Sheet 3249) from Tuckerman’s 144 → FIGURE 18. Thallus morphology of Buellia badia. – A. & B. Neotype specimen of B. badia (on siliceous rock): A. Squamulose thalli growing on and around a Xanthoparmelia species. – B. Close-up of squamulose rosettes growing Xanthoparmelia lobes. – C. & D. Specimen annotated by Imshaug as type of B. turgescens (on decorticated wood): C. Thallus growing among lobes of a Xanthoparmelia . – D. Close-up of bullate thallus areoles growing independently. – E. & F. Lectotype of B. turgescens (on decorticated wood): E. Thallus growing among areoles of a sorediate crust and lobes of a Xanthoparmelia species. – F. Close-up of bullate areoles in close association Xanthoparmelia lobes. – G. & H. Holotype of B. turgescentoides (on siliceous rock): G. Independent bullate areoles. – H. Close-up of the areoles. 145 146 → FIGURE 19. Anatomy of Buellia badia. – A. - D. Neotype specimen of B. badia: A. Exciple. – B. Ascus. – C. Urceolate pycnidium. – D. Conidiophores with bacilliform conidia. – E. - G. Specimen annotated by Imshaug as type of B. turgescens. E. Exciple. – F. Asci. – G. Thallus cross section with phenocortex and thin epinecral layer. H. - J. Lectotype of B. turgescens. H. Exciple. – I. Ascospores. – J. Conidiophores with bacilliform conidia. K. - L. Holotype of B. turgescentoides: K. Exciple. – L. Ascospore. – M. Conidiophores with bacilliform conidia. 147 148 collection at Farlow (FH), Figs. 18C,D, 19E-G]. This annotation was never published and therefore cannot be accepted as a valid lectotypification. We can safely assume that Imshaug first regarded B. turgescens as identical with B. punctata, but later adopted a different concept. In his dissertation, Imshaug describes the difference of the two taxa as follows: “…Most of the material in herbaria called B. turgescens represents a form (cf. var. chloropolia) of B. punctata with a well-developed thallus. The thallus of B. turgescens is composed of large, rounded verrucae with a tendency to become subsquamulose, while the thallus of B. punctata has flattened areoles when well developed…” Imshaug (1951, p. 58). The bullate areoles of the specimen annotated by Imshaug (no. 53.365 ex hb. Tuckerman, FH!) agree well with his description of B. turgescens. Imshaug’s annotation of this specimen as “Buellia punctata” is therefore clearly erroneous. To avoid additional confusion his annotation “Type of Buellia turgescens” must be rejected. Instead we suggest accepting the lectotypification sensu Marbach (2000). Like the specimen annotated by Imshaug, the “Marbach lectotype” has convex, bullate areoles, which are grayish brown to deep brown. The specimen selected by Marbach (2000) therefore also agrees well with Imshaug's (1951) description of B. turgescens. According to Imshaug (1951) the type of Buellia turgescentoides Fink (Fink 739, accession no. 7834, MICH!, Figs. 18G,H, 19K-L) is identical with typical material of B. turgescens. Imshaug (1951) ascribes the Fink description to a misinterpretation of B. turgescens: “… Fink described B. turgescentoides as differing in a “much stronger darker thallus”. Buellia turgescens in Fink’s herbarium, however, is all a form of B. punctata 149 and, consequently, his turgescentoides is the same as Tuckerman’s B. turgescens…” (Imshaug 1951, p. 58). We agree with this assessment. Morphologically the type specimen designated by Fink looks indeed very similar to the lectotype selected by Marbach (2000). Specimens from FH originally annotated by Imshaug as B. turgescentoides show the same morphology as the type material of both B. turgescens and B. turgescentoides. Anatomically all specimens are identical. Buellia turgescentoides is therefore also a synonym of B. turgescens. Imshaug (1951) was convinced that B. turgescens and B. badia should be considered as two different taxa. He made this assessment based on his observations of differences in thallus morphology for the two taxa. However, material annotated by Imshaug (1951) clearly shows that the described morphological variation is present both in specimens he considered to belong to Buellia badia as well as others he annotated as B. turgescens (FH, MSC, MICH). In some of these specimens the thalli are entirely composed of distinctly separate, bullate areoles, whereas other specimens have subsquamulose or even minute squamulose thalli. Even though a few specimens show only one extreme of these growth forms (bullate-areolate vs. squamulose), the complete variation can nevertheless be observed in other specimens. The thalli are therefore equally variable in both taxa and a separation based only on thallus morphology is not justified. Anatomy.—All material examined has the same anatomy. The thallus contains no crystals and is phenocorticate with a thin epinecral layer. The exciple clearly belongs to the aethalea-type sensu Scheidegger (1993) (Fig. 19A,E,H,K). The inner hyphae of the exciple are hyaline and rather thin, parallel in 150 orientation and continuous with a reddish brown hypothecium (leptoclinoidesbrown). The outer hyphae have distinctly swollen cells, which are carbonized throughout by a brown pigment (elachista-brown). Even though it can often be found in other species with the same exciple structure, cinereorufa-green is not present in all of the specimens examined here. The ascospores of B. badia are typical Buellia (= Beltraminea)-type spores with no septum thickening during spore ontogeny (Fig. 19B,F,I,L). Overmature spores show a faint microrugulate ornamentation; spores of other developmental stages are not ornamented. Spore measurements show that width and length of the spores from all specimens fall within the same range [measurements (n = 50) of B. badia neotype: 12.2 ± 1.3 S.D. x 6.7 ± 0.8 s.d.; B. turgescentoides holotype: 11.5 ± 1.9 s.d. x 6.2 ± 0.8 s.d., B. turgescens lectotype sensu Marbach (2000) 11.7 ± 1.0 s.d. x 6.4 ± 0.7 s.d., B. turgescens annotated by Imshaug: 12.3 ± 1.3 s.d. x 6.8 ± 0.8 s.d.]. All specimens have Bacidia-type asci (Fig. 19B,F). Pycnidia are difficult to find on the material. Only a single pycnidium was found and examined on the “Marbach lectotype” (Fig. 19J). The specimen annotated by Imshaug has several pycnidia. Both specimens clearly have bacilliform conidia. Although only a few pycnidia were found on the Fink holotype, bacilliform conidia were also observed in this specimen (Fig. 19K). Substrate, life-strategy and ecology.—Because of its juvenile parasitism, B. badia is quite an unusual member of Buellia. Marbach (2000) argued that the species should therefore be segregated from Buellia and be transferred into Monerolechia Trevisan, a 151 genus erected solely to accommodate parasitic Buellia species. Hafellner (1979), who published a revision of the genus Karschia (parasitic Buellia-like taxa), did not accept the genus Monerolechia. He argued that in Buellia, parasitism only represents a stage during the thallus ontogeny of the species. With age the thalli become independent. No research has yet confirmed that any of the species with juvenile parasitism are obligate parasites. Juvenile parasitism is common also in a variety of other lichen genera. The best known example is probably Diploschistes muscorum, a species, which is initially parasitic on Cladonia-squamules but soon establishes an independent thallus, which even contains a different photobiont than the Cladonia (Friedl 1987). Among the material of B. badia examined here, most specimens show at least some degree of parasitism. In the neotype specimen squamulose thalli can be observed parasitizing a saxicolous Xanthoparmelia (Fig. 18A,B). In close vicinity, some squamules, however, begin to develop independently. In these areas, the thallus squamules grow more closely together, and as a result, become more bullate. These thalli closely resemble the thalli of B. turgescens (both the “Marbach lectotype” and the specimen annotated by Imshaug), which are also largely independent and bullate, rather than squamulose. However, both the “Imshaug” and the “Marbach” specimens show some areas closely associated and partially overgrowing a foliose Xanthoparmelia (Fig. 18D,E,F). In addition, the “Marbach” lectotype also grows closely associated, if not parasitic, among areoles of an unidentified sorediate thallus (Fig. 18E). According to Hafellner (1979), B. badia typically grows on saxicolous foliose species like Neofuscelia and Xanthoparmelia. In the Sonoran Desert the species has also been 152 found on crustose lichen genera such as Acarospora subgen. Xanthothallia and subg. Phaeothallia, and Dimelaena oreina. Specimens on Dimelaena oreina are particularly interesting because Hafellner (1979) described B. imshaugii Hafellner as a parasite on Dimelaena oreina from Canada. Comparison of the type specimen of B. imshaugii (CANL 19372–isotype!) with material of B. badia on Dimelaena oreina from the Sonoran Desert clearly shows the differences of the two species. Buellia imshaugii has a gray areolate thallus, that does not become squamulose. It has larger apothecia and larger ascospores than B. badia. Buellia imshaugii appears to be restricted to higher altitudes or northern latitudes, whereas B. badia is widespread throughout the Northern Hemisphere. In one specimen from Liguria, Italy (M-0061336), both species – B. badia and B. imshaugii – grow together on their host lichen Dimelaena oreina. In general, B. badia thus appears not to be a particularly selective. In addition to the genera of host lichens observed, Rambold & Triebel 1992) mention several other host lichens of this species. Although some specimens can be found without direct association with other lichen thalli, it is possible that these specimens were initially parasitic and that the host lichen disappeared as a result of the parasitism. Buellia badia is most commonly found on rock. Nevertheless, it is not unusual to observe some specimens on dry, decorticated wood impregnated with mineral dust. These specimens were previously referred to as B. turgescens. Both the “Imshaug” and the “Marbach” specimens grow on decorticated wood. They are closely associated with a Xanthoparmelia, a genus also typically found on rock substrates. In contrast, the type specimen of B. turgescentoides grows on rock, not distinctly associated with another 153 lichen thallus. It may not be unusual that a juvenile parasitic lichen will establish on its host even if this host does not grow on its typical substrate. ACKNOWLEDGMENTS We are grateful to Scott Bates, Dr. Philip May and an anonymous reviewer for reviewing the manuscript. Loans from the following herbaria to ASU are greatly appreciated: BM, CANL, COLO, FH, H, hb. Scheidegger, L, M, MICH, MSC, SBBG, UPS, W. The study was supported by a Research Grant in Plant Systematics from the International Association of Plant Taxonomists (IAPT) and National Science Foundation Awards to ASU (DEB-0103738, DEB-9701111) and MSU (DBI-0237401). LITERATURE CITED CULBERSON, C. F. & H. KRISTINSSON. 1970. A standardized method for the identification of lichen products. Journal of Chromatography 46: 85-93. CULBERSON, C. F. & A. JOHNSON. 1982. Substitution of methyl tert.-butyl ether for diethyl ether in standardized thin-layer chromatographic method for lichen products. Journal of Chromatography 238: 438-487. EGAN, R. S. 2001. Long-term storage of TLC data. Evansia 18: 19-20. FRIEDL, T. 1987. Thallus development and phycobionts of the parasitic lichen Diploschistes muscorum. Lichenologist 19: 183-191. GREUTER, W., J. MCNEILL, F. R. BARRIE, H.-M. BURDET, V. DEMOULIN, T. S. FILGUEIRAS, D. H. NICOLSON, P. C. SILVA, J. E. SKOG, P. TREHANE, N. J. TURLAND & D. L. HAWKSWORTH (eds.). 2000. International Code of Botanical 154 Nomenclature (St Louis Code), adopted by the Sixteenth International Botanical Congress St Louis, Missouri, July-August 1999. Regnum Vegetabile 138: 1-474. HAFELLNER, J. 1979. Karschia. Revision einer Sammelgattung an der Grenze von lichenisierten und nichlichenisierten Ascomyceten. Beiheft zur Nova Hedwigia 62: 1-248. IMSHAUG, H. A. 1951. The Lichen-forming Species of the Genus Buellia in the United States and Canada. University Microfilms. Ann Arbor, Michigan. Publication No. 2607. MARBACH, B. 2000. Corticole und lignicole Arten der Flechtengattung Buellia sensu lato in den Subtropen und Tropen. Bibliotheca Lichenologica 74: 1-384. MEYER, B. & C. PRINTZEN. 2000. Proposal for a standardized nomenclature and characterization of insoluble lichen pigments. Lichenologist 32: 571-583. MIETZSCH, E., H. T. LUMBSCH & J. A. ELIX. 1994. WINTABOLITES (Mactabolites for Windows). Users manual and computer program. University Essen, Essen. NORDIN, A. 2000. Taxonomy and phylogeny of Buellia species with pluriseptate spores (Lecanorales, Ascomycotina). Symbolae Botanicae Upsalienses 33: 1-117. NYLANDER, W. 1858 ('1857'). Énumération générale des Lichens, avec l'indication sommaire de leur distribution géographique. Mém. Soc. Sci. Nat. Cherbourg . 5: 85-146. ORANGE, A., P. W. JAMES & F. J. WHITE. 2001. Microchemical Methods for the Identification of Lichens. British Lichen Society, London. 155 RAMBOLD, G. & D. TRIEBEL. 1992. The inter-lecanoralean Associations. Bibliotheca Lichenologica 48: 1-201. SCHEIDEGGER, C. 1993. A revision of European saxicolous species of the genus Buellia De Not. and formerly included genera. Lichenologist 25: 315-364. SHEARD, J. W. & P. F. MAY. 1997. A synopsis of the species of Amandinea (lichenized Ascomycetes, Physciaceae) as presently known in North America. The Bryologist 100: 159-169. SHREVE, F. & I. L. WIGGINS. 1964. Vegetation and Flora of the Sonoran Desert. Stanford University Press, Stanford. TUCKERMAN, E. 1872. Genera Lichenum: An arrangement of the North American lichens. . Amherst, Massachusetts. VOBIS, G. 1980. Bau und Entwicklung der Flechten-Pycnidien und ihrer Conidien. Bibliotheca Lichenologica 14: 1-141. VOBIS, G. & D. L. HAWKSWORTH. 1981. Conidial lichen-forming fungi, pp. 245-273. In COLE, G. T. & B. KENDRICK (eds.), 1: Biology of Conidial Fungi. Academic Press, New York. WHITE, F. J. & P. W. JAMES. 1985. New guide to microchemical techniques for the identification of lichen substances. British Lichen Society Bulletin 57: 1-41. 156 PUBLICATION 3 (The Bryologist 107, 2004): Buellia saurina belongs to the genus Rhizocarpon (Rhizocarpaceae, Lichenized Ascomycetes) FRANK BUNGARTZ Arizona State University Lichen Herbarium, School of Life Sciences, P. O. Box 87 4501, Tempe, AZ 85287 – 4501, U.S.A.; e-mail: frank.bungartz@asu.edu ALAN M. FRYDAY Michigan State University, Herbarium, Department of Plant Biology, 166 Plant Biology, East Lansing, MI 48824 – 1312, U.S.A.; e-mail: fryday@msu.edu Abstract. In 1971, W.A. Weber described Buellia saurina from aeolian sandstone in the Dinosaur National Monument in Utah, U.S.A. In his description Weber mentioned several characters that are rather unusual for the genus Buellia, especially a lemon yellow thallus and a violet-brown hymenium. Careful re-examination of material at ASU and the holotype from COLO demonstrates that the species does not belong to Buellia, but needs to be transferred to Rhizocarpon. The bright yellow pigment is rhizocarpic acid, a substance not known from Buellia. The asci of the specimens belong to the Rhizocarpontype. Although the young ascospores are two-celled, they frequently become irregularly muriform with age. The violet epihymenial pigment, which can be identified as atra-red, is also unusual in Buellia and currently only known from the taxonomically isolated 157 Buellia vilis Th. Fr. The new combination, Rhizocarpon saurinum (W.A. Weber) Bungartz, is made. During a revision of saxicolous Buellia species with one-septate spores from the Greater Sonoran Desert Region (Nash III et al. 2002), specimen material of Buellia saurina W.A. Weber was examined. This species has been described by Weber (1971) from aeolian sandstone in the Dinosaur National Monument in Utah, U.S.A. Within Buellia the species is rather unique. It forms small, pale lemon, granular-verrucose thalli (Fig. 20A,D). The epruinose black apothecia are immersed between the thallus granules. With standard thin-layer chromatography (Orange et al. 2001) the lemon yellow pigment was identified as rhizocarpic acid, a substance otherwise not known from Buellia. Transverse sections through the apothecium show a diffuse, dark violet pigment throughout the epihymenium and upper parts of the hymenium. The spores are very deeply pigmented and appear blackish olive rather than the deep brown color otherwise typical for ascospores of Buellia. Spore septation can hardly be distinguished because of the extremely dark pigmentation (Fig. 20F). The addition of concentrated HNO3 removes some of this pigment and spore septation can then be studied more easily (Fig. 20B,C). The spores are initially one-septate (Fig. 20B) but soon become irregularly muriform (Fig. 20C). Applying a standardized series of KOH, HNO3, water and HCl to the transverse sections identifies the violet epihymenial pigment as atra-red (sensu Meyer & Printzen 2000). Within Buellia, this pigment is currently only known from Buellia vilis 158 Th. Fr., another unusual species, characterized by a unique exciple type (Scheidegger 1993). Like B. saurina, B. vilis appears to be rather isolated within Buellia. However, although the two species share the same epihymenial pigment, they are not closely related. Buellia vilis is a chasmolithic species i.e., with a barely visible thallus hidden between mineral grains of the substrate. Although barely visible, the thallus reacts strongly with Lugol’s iodine, but lichen substances have so far not been detected. Buellia vilis is restricted to subalpine to alpine habitats. In North America the species is known from the arctic (Thomson 1997b). Scheidegger (1993) described the distribution in Europe as alpine. In the southwestern U.S.A. We are currently aware of only a single specimen. The habitat of this specimen is subalpine (Marsh 116, NEW MEXICO, San Juan County, Barker Arroyo W of Highway 170, SW of La Plata, on sandstone,. 1770 m elev., 36°52'N, 108°15'W). Buellia vilis has a distinct Bacidia-type ascus and therefore clearly belongs to the Physciaceae. Even though it is unique within Buellia, it should be maintained within that genus. The taxonomy of the genus is currently in a state of flux, and at the moment, it is not justified to describe a new genus only to accommodate this unique species. In contrast, B. saurina does not belong to the Physciaceae. In Lugol’s iodine the asci of the species are not even remotely similar to the typical asci of the Physciaceae. All Physciaceae have asci with a central non-amyloid cone and dark amyloid tholus flanks. According to the shape of the central amyloid cone, different ascus types have been distinguished, which are all variations of the same basic ascus structure. The Lecanora- 159 type ascus has tholus flanks, which do not merge at the tip, whereas these flanks converge in the Bacidia- or Biatora-type. In contrast, B. saurina has a distinctly layered ascus tip without a central cone. Instead the ascus is characterized by a strongly amyloid outer cap and a non-amyloid inner layer (Fig. 20E). This ascus is typical for the genus Rhizocarpon. Atra-red is a rather common pigment in the epihymenium of Rhizocarpon. Ascospores of Rhizocarpon are similar to those found in Buellia, but often appear slightly darker and are generally more olive instead of brown. They frequently (but not always) have a thick gelatinous sheath, usually referred to as halo. Rhizocarpic acid is the characteristic pigment of all yellow Rhizocarpon species. Paraphyses in Buellia often branch towards the tip but do not anastomose, as can often be observed in Rhizocarpon. On the basis of these observations, B. saurina is therefore transferred to the genus Rhizocarpon. Species of Rhizocarpon that contain rhizocarpic acid and have a non-amyloid medulla are only found in the R. viridiatrum group (Runemark 1956). However, that group is further characterized by having black granules in the epihymenium, which are not very evident in R. saurinum. The only other species of Rhizocarpon that contain rhizocarpic acid, have a non-amyloid medulla, and have ascospores with similar dimensions and septation as R. saurinum are R. viridiatrum (Wulfen) Körb., and the closely related R. lusitanicum (Nyl. ) Arnold and R. ochrolechiae (Poelt & Nimis) Hafellner. However these species are all lichenicolous (at least initially), usually on Aspicilia cinerea (L.) Körb, Pertusaria spp., or Ochrolechia parella (L.) A. Massal. (Poelt 1990). The two collections of R. saurinum that we have studied occur directly on bare rock (sandstone) 160 with no evidence of directly associated lichens. Rhizocarpon saurinum also has a lower hymenium, and smaller ascospores with fewer septa than these other species. Rhizocarpon atrovirellum (Nyl. ) Zahlbr. has similar ascospores to R. saurinum, but Runemark (1956) considered this to be of little significance and included R. atrovirellum within the synonymy of R. viridiatrum. Consequently, we have no hesitation in considering R. saurinum to be a distinct species rather than a synonym of a previously described one. RHIZOCARPON SAURINUM (W. A. Weber) Bungartz, comb. nov. Buellia saurina W. A. Weber, The Bryologist 74: 190. 1971. TYPE: U.S.A. UTAH. Uintah C., Dinosaur National Monument. Vicinity of Doug Chew Cabin; 2250 m alt.; Blue Mountain Plateau; on loosely cemented Aeolian sandstones of the Mesa Verde Formation; in protected overhang of rimrock ledge; 40°23'N, 109°06'W [Township 4 S, Range 25 E]; 3 June 1956; Weber L-7935 (= S 7935, COLO – holotype!). Thallus (Fig. 20A,D) crustose; granular verrucose to areolate, on an inconspicuous black prothallus; surface matt, pale lemon yellow, roughened, epruinose; phenocorticate with thick epinecral layer of dead cells and calcium oxalate crystals. Apothecia immersed, interspersed between thallus granules; lecideine; proper margin thin, becoming excluded with age; disk and margin black, epruinose, soon becoming convex; exciple narrow (Fig. 20F,G), poorly differentiated, inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), similar in structure and orientation to 161 paraphyses, often ± reduced and transient with dull reddish brown hypothecium (leptoclinoides-brown, textura intricata); outer excipular hyphae parallel (textura oblita), cells moderately swollen and usually strongly carbonized with various amounts of strongly violet (HNO3+ purple) pigment (atra-red), pigmentation continuous with epihymenium; diffusing into upper parts of otherwise hyaline hymenium, paraphyses not inspersed, simple to moderately branched, occasionally anastomosing with one another, apically swollen, with deep violet (HNO3+ purple) pigment cap (atra-red). Asci 8-spored, clavate, Rhizocarpon-type (Fig. 20E); Ascospores (Fig. 20B,C) broadly ellipsoid to globose, not constricted, with obtuse ends, not curved, (12.0–)14.8–[17.0]–19.1(–25.0) x (10.0–)10.8–[12.2]–13.6(–15) µm (n = 50) (measurement format according to Nordin 2000), deep olive black, one-septate to irregular muriform, septa narrow, not thickening during spore ontogeny, with indistinct gelatinous sheath (inconspicuous halo), old spores microrugulate. Pycnidia not found. Chemistry.—Rhizocarpic acid. Thallus spot test reactions negative, cortex and medulla UV-. Medulla I–. Substrate and ecology.–Currently known only from sheltered sandstone outcrops in sagebrush and chaparral. Distribution.—Known from Utah and Colorado, U.S.A. Additional material examined.—U.S.A. COLORADO. Moffat Co., E side of highway 13, ca. 26 km N of Axial; ca. 1875 m elevation; in sagebrush community; W-facing hill; on sandstone outcrops; 40°24'N, 107°38'30''W; Ryan 20658 (ASU). 162 → FIGURE 20. Morphology and anatomy of Rhizocarpon saurinum (L-7935 = S 7935, COLO – holotype!).—A. Granular verrucose thallus.—B. Young, one-septate ascospore (in concentrated HNO3).—C. Mature, irregularly muriform ascospores (in concentrated HNO3).—D. Close-up of the granular verrucose thallus and apothecia.—E. Two immature asci in Lugol’s solution show a distinct amyloid cap and an otherwise pale tholus (Rhizocarpon-type).—F. Cross section through the proper exciple in water: In the outer exciple, separate hyphae cannot be distinguished because of a deep violet carbonization of the cells by the pigment atra-red. The inner hyphae are similar in structure and orientation to the paraphyses. These hyphae are continuous with a deep brown (leptoclinoides-brown) hypothecium of irregularly interwoven hyphae (textura intricata).—G. Cross section through the proper exciple in HNO3: The carbonization has partially been dissolved and the structure of the outer exciple can be distinguished as swollen cells of loosely interwoven mesodermatous hyphae (textura oblita). 163 164 ACKNOWLEDGMENTS We are grateful to Dr. Thomas H. Nash III, Scott Bates (ASU) and two anonymous reviewers for reviewing the manuscript, and the curator of COLO for the loan of the holotype of Buellia saurina. The study was supported by a Research Grant in Plant Systematics from the International Association of Plant Taxonomists (IAPT) and National Science Foundation Awards to ASU (DEB-0103738, DEB-9701111) and MSU (DBI-0237401) . LITERATURE CITED MEYER, B. & C. PRINTZEN. 2000. Proposal for a standardized nomenclature and characterization of insoluble lichen pigments. Lichenologist 32: 571-583. NASH III, T. H., B. D. RYAN, C. GRIES & F. BUNGARTZ (eds.). 2002. Lichen Flora of the Greater Sonoran Desert Region 1: 1-532. NORDIN, A. 2000. Taxonomy and phylogeny of Buellia species with pluriseptate spores (Lecanorales, Ascomycotina). Symbolae Botanicae Upsalienses 33: 1-117. ORANGE, A., P. W. JAMES & F. J. WHITE. 2001. Microchemical Methods for the Identification of Lichens. British Lichen Society, London. POELT, J. 1990. Parasitische Arten der Flechtengattung Rhizocarpon: eine weitere Übersicht. Mitteilungen der Botanische Staatssammlung München 29. RUNEMARK, H. 1956. Studies in Rhizocarpon. I. Taxonomy of the yellow species in Europe. Opera botanica 2: 1-152. 165 SCHEIDEGGER, C. 1993. A revision of European saxicolous species of the genus Buellia De Not. and formerly included genera. Lichenologist 25: 315-364. THOMSON, J. W. 1997. American Arctic Lichens. 2. The Microlichens. University of Wisconsin Press, Madison. WEBER, W. A. 1971. Four new species of Buellia (Lichenes) from western North and South America. The Bryologist 74: 185-191. 166 PUBLICATION 4 (Bibliotheca Lichenologica 88, 2004): Buellia subalbula (Nyl.) Müll. Arg. and B. amabilis de Lesd., two species from North America with one-septate ascospores: A comparison with Buellia [“Diplotomma”] venusta (Körb.) Lettau. FRANK BUNGARTZ and THOMAS H. NASH III School of Life Sciences, Arizona State University, P.O. Box 87 4501, Tempe, AZ 85287-4501, U.S.A. Abstract: This study compares three saxicolous Buellia species with ± pruinose apothecia and rimose thalli rich in calcium oxalate. Buellia subalbula (Nyl.) Müll. Arg. is a new record to North America. It is a common crustose lichen on coastal rocks of southern California (USA) and northern Baja California (Mexico). The species was first described from Cabo Negro in Angola, along the west coast of Africa. It has also been reported from the Negev Desert in Israel, Saudi Arabia and Australia. In Europe the species has previously been referred to Buellia maritima (A. Massal.) Bagl., a taxon which is not identical with B. stellulata (Taylor) Mudd. Buellia amabilis de Lesd. is a morphologically similar species known only from pebbles at the type locality in Acatzingo, central Mexico. In comparison with B. subalbula, B. amabilis has consistently larger apothecia, a more distinctly developed exciple, and larger ascospores with a conspicuous rugulate ornamentation. Both species have one-septate spores. In contrast, the widely distributed B. venusta (Körb.) Lettau has pluriseptate spores and is therefore 167 often treated as a member of the genus Diplotomma Flot. The similar thallus morphology and the presence of large amounts of calcium oxalates suggest an affinity of B. subalbula and B. amabilis to the “Diplotomma-group”, even though both species have one-septate ascospores. Ultrastructural studies of the spore wall show a considerably thickened perispore in B. amabilis and B. venusta but a thin perispore in B. subalbula. The spore wall is ornamented in spores of B. amabilis and B. venusta but ornamentation is absent from B. subalbula spores. Nevertheless, it is presently not clear if the genus concept of Diplotomma needs to be emended to include species with one-septate spores such as B. amabilis but not B. subalbula. The strongly rugulate ornamentation of B. amabilis spores is caused by a coarsely fractured perispore whereas B. venusta has microrugulate spores as a result of a finely fissured perispore. As part of a taxonomic revision of saxicolous species of the genus Buellia de Not. with one-septate ascospores, Buellia subalbula (Nyl.) Müll. Arg. was discovered among herbarium specimens from the Sonoran Desert Region. In North America the species is common along the coast of southern California, USA and Baja California, Mexico. Buellia subalbula has previously not been reported from North America. The species was generally overlooked and commonly misidentified as B. retrovertens Tuck., a synonym of B. dispersa A. Massal. (Bungartz et al. 2002). The confusion of the two taxa can be attributed to considerable morphological variation in B. dispersa. Even though thalli of B. subalbula are distinctly rimose, the species was not previously distinguished from areolate to subsquamulose thalli of B. dispersa. Imshaug, however, must have 168 recognized B. subalbula, which he annotated as “B. subrinodinoides” (Lich. Exs. no. 4 ex hb. Hasse, FH), a name never validated in a publication. The Latin diagnosis of B. amabilis suggested that this species, described by Bouly de Lesdain from Acatzingo in Mexico, is very similar to B. subalbula and possibly the same taxon. We therefore examined type material from B. amabilis, but discovered that this species is distinctly different from B. subalbula. Thallus morphology of B. subalbula and B. amabilis is very similar to some species with pluriseptate spores, namely B. venusta (Körb.) Lettau. Species of Buellia with pluriseptate spores have commonly been segregated as Diplotomma Flot. This genus is currently accepted by several checklists and treatments: Singh & Awasthi (1989 - India), Purvis (1992 - British Isles), Nimis (1993 - Italy), Esslinger (1997 - U.S.A. & Canada), Randlane & Saag (1999 - Estonia). Other authors include species with pluriseptate spores within Buellia: Singh (1964 - India), Poelt (1969 - Europe), Ozenda & Clauzade (1970 France), Wirth (1994; 1995 - Germany),Scholz (2000 - Germany), Clauzade & Roux (1985 - Europe), Santesson (1993 - Sweden), Thomson (1997b - American Arctic). When Flotow (1849a; 1849b; validly published in 1850) described Diplotomma, he suggested a “double exciple” as the most important character to distinguish the new genus from Lecidea. His description of the genus did not consider spore septation, but Massalongo (1852) subsequently emphasized pluriseptate ascospores to separate Diplotomma from Buellia. The genus concept was further clarified when Körber (1860) excluded Diploicia canescens and Rhizocarpon geographicum, and Clements & Shear (1931) selected Buellia alboatra as the type species of the genus. Szatala (1955) included 169 all Buellia species with pluriseptate spores in Diplotomma. Purvis (1992) suggested a more narrow concept and argued that only species with structurally different septa should be included in Diplotomma. Nordin (1996), however, demonstrated that septum formation in the Physciaceae generally follows the same pattern and therefore did not accept Diplotomma solely on the basis of ascospore septation. As the result of a recent cladistic analysis of classical data Nordin (2000) suggested confining Diplotomma s. str. to a group of species characterized by pluriseptate ascospores, thalli rich in calcium oxalates, apothecia frequently covered with pruina, and ascospores characterized by a conspicuously thickened perispore. Nordin (2000) emphasized that these characters must be regarded as parallelisms. Although they appear to be present in characteristic combination only within Diplotomma s.str., they can independently also be found in other groups of Buellia s.l. Therefore, Nordin (2000) hesitated to recognize the genus and did not provide an emended, formal description of Diplotomma. In this publication we examine species with one-septate ascospores, which display some characteristics of the “Diplotomma-group” sensu Nordin (2000). We compare B. subalbula, B. amabilis and B. venusta, and demonstrate that only B. amabilis might be included in the “Diplotomma-group” sensu Nordin (2000). METHODS All specimens were examined with light microscopy using hand- and cryosections. Both conventional bright field microscopy (BF) as well as differential interference contrast (DIC) was used to examine the specimens. Selected specimens were also studied with scanning electron (SEM) and transmission electron microscopy (TEM) according to 170 a protocol described in detail by Bungartz et al. (2002). To improve dehydration and infiltration this protocol has been modified using a laboratory microwave as follows: specimens in Eppendorf vials were incubated in a 340 ml water bath with a load controller of 600 ml water calibrated at 20 °C and a temperature controller calibrated at 37 °C. Power output of the microwave was set at 50% (medium). Using these settings specimens were dehydrated in a series of 50, 70 and twice in 100% ethanol for 5 min each. Subsequently, specimens were infiltrated at the same settings in a series of 50, 70 and two times 100% resin - ethanol mixture for 20 min each. Representative specimens of the three species were analyzed with powder X-ray diffraction (XRD, Fig. 24), to determine if large amounts of calcium oxalates were present in the thallus. As an unspecific indicator for the presence of oxalates, 10% H2SO4 was added to thallus cross sections. This results in the formation of characteristic sulfate needles if oxalates are present (Fig. 21E). All specimens were spot tested and routinely examined with standardized thin layer chromatography (Culberson & Kristinsson 1970; Culberson & Johnson 1982; White & James 1985; Orange et al. 2001). TLC-Plates were interpreted with the computer program WINTABOLITES (Mietzsch et al. 1994), and scanned for permanent record (Egan 2001). Spores measurements are given according to Nordin (2000). Pigment names follow Meyer & Printzen (2000). RESULTS BUELLIA SUBALBULA (Nyl.) Müll. Arg. Revue Mycolog. 2: 79. 1880. 171 Lecidea subalbula Nyl. Bull. Soc. Linn. Normandie 2(2): 516. 1867. TYPE: ANGOLA. BENGUELA. Black Mountains, near Namibe (formerly Mossâmedes), in a desert region, widely distributed around Capo Negro, on calcareous rock, 15°41'00''S, 11°56'00''E [original label data: Benguella, Montes negros, ad rupes calcareas, propre Mossâmedes, in regione sterilissima; magis evoluta in Capo Negro] coll. date 1859, Nylander 9319a (H-NYL, lectotype selected here), Nylander 9319b (H-NYL, isolectotype). Note: Both type specimens are very small fragments. No. 9319a and 9319b appear to be part of the same collection designated by Nylander as the type specimen. The reason why these two fragments have been separated and assigned collection numbers 9319a & b respectively, is not evident from the two specimens. For clarification the larger specimen (9319a) with less than ten intact apothecia is selected here as a lectotype. The second fragment with a single apothecium thus becomes an isolectotype. Buellia maritima (A. Massal.) Bagl., in A. Massal. Schedul. Critic. 8: 150. 1856.— Catolechia maritima A. Massal., Framm. Lich.: 22. 1855. TYPE: ITALY. LIGURIA. on rock. [original label data: Ad saxa in Liguria propre Genuam] A. Massal. Lich. exs. Ital. no. 271 [Scheidegger (1993): “part of this number is confused with no. 272, Buellia dispersa”] (M-0061351!, TO – isotypes). Note: Scheidegger (1993) suggested that B. maritima was a synonym of B. stellulata (Taylor) Mudd. Although we have not studied type material from TO, the material from M is identical with B. subalbula. B. maritima is thus a synonym of B. subalbula. Buellia subrinodinoides Imshaug ined. 172 U.S.A. California: Santa Monica Range, no. 4. Lichenes Exsiccati ex herbario Hasse, reclicti, distributed by C. C. Plitt, Sullivant Moss Society (FH, original identification as B. alboatra var. saxicola Fr.; annotated by Imshaug 1952 as Buellia subrinodinoides Imshaug ined.). Thallus (Fig. 21A–C) crustose; rimose, thin thalli appearing continuous to infrequently cracked, thick thalli becoming rimose-areolate but areoles poorly delimited; usually growing in distinct circular patches, subeffigurate, several thalli often confluent; thalli usually delimited by an indistinct whitish or grayish darkened or distinctly blackened prothallus; surface matt, chalky white to grayish, roughened, whitish pruinose; phenocorticate with thick epinecral layer of dead cells and calcium oxalate crystals; entire thallus filled with an abundance of calcium oxalate (typically both the monohydrate weddellite and the dihydrate whewellite). Apothecia immersed, becoming adnate to sessile; lecideine but rarely surrounded with outer ring of poorly differentiated thalline material (thalline collar, Fig. 21B); proper margin becoming excluded with age; disk black, pruinose or not, plane, becoming convex with age; exciple of aethalea-type (Fig. 21F,G) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), similar in structure and orientation to the paraphyses, often ± reduced and transient with the dull reddish brown hypothecium (leptoclinoides-brown, textura intricata); outer excipular hyphae parallel (textura oblita), cells moderately swollen and usually strongly carbonized with various amounts of brown and aeruginose (HNO3+ violet) pigments (cf. elachista-brown & cinereorufa-green), 173 pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed, paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown) and a diffuse aeruginose pigment (HNO3+ violet, cinereorufagreen). Asci 8-spored, clavate, Biatora-type (Fig. 21D); Ascospores (Fig. 21I,K) narrowly-oblong to ellipsoid, not constricted, with obtuse ends, not curved, (8.0–)10.4– 11.6–12.8(–15.0) × (4.0–)5.1–5.8–6.5(–8.0) µm (n = 615), one-septate, proper septum narrow, not thickening during spore ontogeny, with septal pore canal, simple pore and undifferentiated pore plug; ornamentation absent (or inconspicuous, not distinguishable with DIC); spore wall differentiated into smooth, thin perispore (< 0.10 µm), narrow intermediate layer (< 0.10 µm), thick proper spore wall (0.30–0.50 µm) and moderately thickened endospore (0.10–0.18 µm). Pycnidia rare; globose, unilocular; at maturity almost entirely occupied by densely branched conidiophores; conidiogenous cells intercalary and terminal (conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type sensu Vobis (1980) and Vobis & Hawksworth (1981); conidia (Fig. 21H) simple, bacilliform, 2.5–4.0 µm × 1.0–1.5 µm (n = 30). Chemistry.—Norstictic acid (major), connorstictic acid (traces), rarely secondary metabolites absent. Thallus typically K+ yellow to red, forming characteristic needleshaped crystals (if observed with the compound microscope), P+ yellow, C–, KC–, CK– (all reactions usually well pronounced, rarely absent). Medulla I– (important: always test with the compound microscope, positive reactions with Lugol’s iodine can be very weak!). Note: John A. Elix (University of Canberra) annotated the type specimen as follows: “atranorin, stictic acid (major), constictic acid, unknown depside (minor) 174 Methods: TLC (C), HPLC”. Analysis of the same specimen fragment as annotated by Elix but did not confirm these substances. In contrast, our analysis clearly demonstrated the presence of norstictic and connorstictic acid in the type material as consistent with the rest of the specimens examined. Substrate and ecology.—Known only from rock substrates with traces of calcium carbonate (limestones, carbonate-rich sandstones). Often found at nutrient enriched sites. Often growing associated with B. venusta. Distribution.—Presently known from south-western North America, west Africa, the Middle East and Australia (see specimens examined). All specimens examined are from coastal areas at low elevations. In contrast, Abu-Zinada et al. (1986) and Bokhary (1993) report B. subalbula from higher altitudes of Saudi Arabia. We have not examined any material from these localities. Two specimens from Yemen (Schultz 14151f, 14015e), also collected at high altitudes, are very similar but not identical with B. subalbula (see discussion). Ghazanfar & Gallagher (1998) report the species from coastal fog areas in the Sultanate Oman. One specimen at ASU collected in the Negev Desert of Israel [Galun 26(1)] matches well with the North American material. The species may also be common along the coasts of Mediterranean Europe, where it was probably misidentified as B. stellulata. Scheidegger (1993) suggested that B. maritima is a synonym of B. stellulata. Buellia maritima is, however, identical with B. subalbula. It has the same rimose thallus with large amounts of calcium oxalates, identical apothecial anatomy and spores of the same size and structure. All specimens of B. maritima from M contain norstictic acid, unlike B. stellulata, which is characterized by atranorin, confluentic and 175 2'-O-methylperlatolic acid. Buellia stellulata has a distinctly areolate thallus, with a smooth, epruinose surface and contains few, if any calcium oxalate crystals. Specimens examined.—U.S.A. CALIFORNIA. Los Angeles Co. Lich. Exsic. no. 4 ex hb. Hasse (FH, ASU; distributed as B. alboatra var. saxicola Th. Fries), ex herb. Hasse s.n. (FH); Nash 34109 (ASU); San Francisco Co. Sigal s.n., (ASU); Santa Barbara Co. Nash 32674, 41126, 41100, 41201, 41222, 41272, 41270, 41323, Ryan 31029, 31054, Sheard 5054b (ASU); Wetmore 73630, 73685 (MIN), Printzen 44, 48, 51, 59, 74 (hb. Printzen); Sheard 5054b-A (SASK); Ventura Co. Marsh 8002, Nash , 32675, 38658a, 38658c, 38659, 38668, 38735, 38898, 38901a, 38901c, 41398 (ASU); Bratt 7881b, 7981, 11997, 12000, Fusaro s.n., Tucker 33692a, c, 33727, 33792a, 33813 (SBBG); Foreman L-41680, L44268, L-44283, Weber L-88975 (COLO). MEXICO. BAJA CALIFORNIA. Wiggins 16196 (DS, now part of CAS); Bungartz 2294 (hb. Bungartz, ASU); Marsh 7282, Nash 33994, 34597, 40137 (ASU). BAJA CALIFORNIA SUR: Nash 29526, 29605, 33853, 33953, 33954, 34606, 39797 (ASU), Moberg 10358 (UPS). ANGOLA. BENGUELA: Nylander 9319a, b (H). ISRAEL. NEGEV DESERT. Galun 26(1) (ASU). AUSTRALIA. SOUTH AUSTRALIA PROVINCE. Nash 19887 (ASU). ITALY. LIGURIA. Lich. Selecti Exsic. no. 25 Vězda (M0061348, M-0061347), Cevasco s.n. (M-0061349, M-0061349, M-0061350), hb. Kayser ex Massal. Lich. exs. Ital. no. 271 (M-0061351). Specimens similar but not identical to B. subalbula: YEMEN. HADRAMAUT GOUVERNORATE: Schultz 14151f, 14015e (hb. Schultz, duplicates in hb. Bungartz). 176 BUELLIA AMABILIS de Lesd. Lich. du Mexique: 27. 1914. TYPE: MEXICO. Puebla: Acatzingo, on volcanic rocks (pebbles embedded in soil), 18°59'00''N, 97°47'00''W, coll. date 1907, Amable 4196 [original label data: Puebla: Acatzingo, sur les roches volcaniques (tépétate), Frère Amable no. 4196] (US, lectotype selected here) Note: Lectotypefication of type material designated by Maurice Bouly de Lesdain is necessary because it can generally be assumed that his entire holotype collection, originally located in Dunkerque (France) has been destroyed during the Second World War. 177 TABLE 4. Diagnostic differences in B. amabilis, B. subalbula and B. venusta. species spore septation spore ornamentation spore perispore exciple anatomy exciple color exciple hyphal texture chemistry Buellia amabilis oneseptate rugulate thick, fractured outer exciple fuscous brown (not carbonized) hyaline pale reddish brown thin, parallel no substances aeruginose & fuscous brown (carbonized) hyaline deep reddish brown swollen, parallel inner exciple hypothecium Buellia subalbula oneseptate absent (psilate) thin, smooth outer exciple inner exciple hypothecium Buellia venusta pluriseptate microrugulate thick, fissured outer exciple inner exciple hypothecium fuscous brown (not carbonized) hyaline deep reddish brown inter-woven inter-woven norstictic acid thin, parallel inter-woven thin, parallel thin, parallel inter-woven no substances or with norstictic acid 178 → FIGURE 21. Buellia subalbula.—A. Thin thallus on calcium carbonate rich sandstone. A fimbriate, white prothallus delimits the outline of the thallus (Bratt 7981).—B. Thick thallus on limestone without distinct prothallus. Apothecia are slightly pruinose outlined by a thin thalline collar (Nash 33853).—C. Thalli on small limestone pebbles, distinctly delimited by a distinct black prothallus (Ryan 31029).—D. Light micrograph (DIC) of a Biatora-type ascus stained with Lugol’s iodine (Nash 32675).—E. Calcium sulphate crystals forming in the medulla after adding 10% sulphuric acid (Bratt 7891).—F. Light micrograph (DIC) of the aethalea-type exciple (Nash 32675).—G. SEM micrograph of the aethalea-type exciple (Bungartz 2291).—H. Light micrograph (DIC) of conidiophores and conidia (Nash 33853).—I. TEM micrograph of a spore (the ultrastructure of the cytoplasm is not preserved, Bungartz 2291).—J. TEM micrograph of the spore wall ultrastructure; (1) thin, smooth perispore; (2) thin intermediate layer; (3) thick proper spore wall; (4) endospore (Bungartz 2291). 179 180 → FIGURE 22. B. amabilis (all images from the type specimen, Amable 4196).—A. Thallus on calcium carbonate rich pebble.—B. Close-up of the pruinose apothecia (note the larger size compared to apothecia in Fig. 4.1.A-C and 4.3A,B).—C. Light micrograph (DIC) of the exciple.—D. Light micrograph (DIC) of a coarsely rugulate ascospore.—E. TEM micrograph of an ascospore (the ultrastructure of the cytoplasm is not preserved).— F. TEM micrograph of the spore wall ultrastructure. (as) ascus wall, (m) mucilaginous sheath, (1) thick, fractured perispore; (2) thin intermediate layer; (3) thick proper spore wall; (4) endospore. 181 182 → FIGURE 23. B. venusta.—A. Thallus growing on limestone (Nash 38902).—B. The thallus of B. venusta (v, Bratt 7981a) growing besides B. subalbula (s, Bratt 7981b). A narrow black prothallus line barely distinguishes the two thalli (arrow).—C. Light micrograph (DIC) of the exciple (Nash 38690).—D. Light micrograph (DIC) of a pycnidium (Wetmore 17095).—E. Light micrograph (DIC) of conidiophores (Wetmore 17095).—F. Light micrograph (DIC) of conidia (Wetmore 17095).—G. Light micrograph (DIC) of ascospores at different stages during spore ontogeny; (y) young, hyaline ascospore with a single median septum (arrow); (p) premature ascospore with additional septum formation (arrow), dark spore wall and the development of a microrugulate ornamentation (Nash 32690). Note: These spores also show the formation of a few longitudinal septa, which occasionally can be found in pluriseptate spores.—H. TEM micrograph of an ascospore (the ultrastructure of the cytoplasm is not preserved, Bratt 7981a).—I. TEM micrograph of the spore wall differentiation; (as) ascus wall, (m) mucilaginous sheath, (1) thick, fissured perispore; (2) thin intermediate layer; (3) thick proper spore wall; (4) endospore (Bratt 7981a). 183 184 → FIGURE 24. Powder X-ray diffraction spectrum of the thallus of B. venusta (Nash 38902). 185 186 Thallus (Fig. 22A,B) crustose; rimose, thickened, becoming rimose-areolate; usually growing in distinct circular patches, subeffigurate, sometimes confluent, usually delimited by an indistinct whitish or grayish darkened or distinctly blackened prothallus; surface matt, gray to whitish gray, roughened, whitish pruinose; phenocorticate with thick epinecral layer of dead cells and calcium oxalate crystals; entire thallus filled with an abundance of calcium oxalate (typically both the monohydrate weddellite and the dihydrate whewellite). Apothecia (Fig. 22B) immersed, becoming adnate to sessile; lecideine but rarely surrounded with outer ring of poorly differentiated thalline material (thalline collar); proper margin soon becoming excluded with age; disk black, usually pruinose, plane, soon becoming convex; exciple similar to the leptoclinoides-type (Fig. 22C) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous interwoven (± textura intricata), continuous and similar in structure with the pale reddish brown hypothecium (leptoclinoides-brown); outer excipular hyphae thin-walled, parallel (textura oblita), usually strongly carbonized with various amounts of a fuscous brown pigment (cf. elachista-brown), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed, paraphyses simple to moderately branched, apically swollen, with brown pigment cap (cf. elachista-brown). Asci 8-spored, clavate, Bacidia-type; Ascospores (Fig. 22E,F) oblong to ellipsoid, not or slightly constricted, with obtuse ends, usually not curved, (18.0–)18.7–20.7–22.7(–26.0) × (9.0–)10.2–11.2–12.1(–13.0) µm (n = 50), one-septate, proper septum narrow, not thickening during spore ontogeny, with septal pore canal, simple pore and undifferentiated pore plug; ornamentation distinct (conspicuously microrugulate to 187 rugulate in DIC); spore wall differentiated into thick, conspicuously fractured perispore (c. 0.50–0.70 µm), indistinct intermediate layer, thick proper spore wall (0.30– 0.50 µm) and thickened endospore [0.10–0.20 (–0.30) µm]. Pycnidia not found. Chemistry.—no substances found by TLC! Thallus spot test reactions negative. Medulla I- Note: Only the type material was available for examination and specimens with small amounts of secondary metabolites may occur. Substrate, ecology and distribution.—Only known from the type locality. Specimens examined.—MEXICO. PUEBLA. Only the type material (Amable 4196) was available. BUELLIA VENUSTA (Körb.) Lettau. Hedwigia 52: 244. 1912. Diplotomma venustum Körb. Syst. Lich. German.: 179. 1860. Note: for further synonyms and a discussion of the taxonomy see Nordin (1996; 2000). Thallus (Fig. 23A,B) crustose; rimose, thin thalli appearing continuous to infrequently cracked, thick thalli becoming rimose-areolate but areoles poorly delimited; usually growing in distinct circular patches, subeffigurate, several thalli often confluent; thalli usually delimited by an indistinct whitish or grayish darkened or distinctly blackened prothallus; surface matt, chalky white to grayish, roughened, whitish pruinose; phenocorticate with thick epinecral layer of dead cells and calcium oxalate crystals; entire thallus filled with an abundance of calcium oxalate (typically both the monohydrate weddellite and the dihydrate whewellite). Apothecia immersed, becoming adnate to sessile; lecideine but often surrounded with outer ring of poorly differentiated thalline 188 material (thalline collar); proper margin becoming excluded with age; disk black, pruinose or not, plane, becoming convex with age; similar to the aethalea-type (Fig. 23C) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), similar in structure and orientation to the paraphyses, often ± reduced, continuous with the reddish brown pigmented hypothecium (leptoclinoides-brown); outer excipular hyphae parallel (textura oblita), only the outer cells carbonized with various amounts of brown pigments (cf. elachista-brown), not considerably swollen, pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed, paraphyses simple to moderately branched, apically swollen, with a dull brown pigment cap (cf. elachista-brown). Asci 8-spored, clavate, Bacidia-type; Ascospores (Fig. 23G–I) broadly-oblong to ellipsoid, not constricted, with obtuse ends, often curved, (12.5–)14.0–15.7–17.3(–17.5) × (5.0–)7.1–8.7–10.1(–12) µm (n = 24) (compare measurements with Nordin 2000), three-septate, proper septa narrow, not thickening during spore ontogeny, with septal pore canal, simple pore and undifferentiated pore plug, rarely with additional longitudinal septa; spore ornamentation distinct (conspicuously microrugulate to rugulate in DIC); spore wall differentiated into finely fissured, thick perispore (0.40–0.65 µm), narrow, conspicuous intermediate layer (0.03–0.05 nm), moderately thickened proper spore wall (0.15–0.20 (–0.30) µm) and moderately thickened endospore (0.10–0.15 µm). Pycnidia (Fig. 23D) rare; globose, unilocular; wall at maturity densely covered by short, rarely branched conidiophores (Fig. 23E) with terminal conidiogenous cells (conidiophores similar to type V sensu Vobis 1980, but conidiogenous cells on considerably shortened hyphae); pycnidial ontogeny 189 similar to the Umbilicaria-type sensu Vobis (1980) and Vobis & Hawksworth (1981); conidia (Fig. 23F) simple, bacilliform to fusiform, 7.0–12.0 µm long (n = 20). Chemistry.—Secondary metabolites absent or rarely with norstictic acid (major), connorstictic acid (traces). Thallus typically K+ yellow to red, forming characteristic needle-shaped crystals (if observed with the compound scope), P+ yellow, C–, KC–, CK– (all reactions usually well pronounced, rarely absent). Medulla I–. Substrate and ecology.—Locally very abundant on a wide variety of calciferous rocks including man-made substrates such as cement, mortar, concrete or even brickwork. Often at nutrient-rich sites. Distribution.—Widely distributed throughout the Northern Hemisphere in North America, Europe, northern Africa and Asia. Specimens examined.—U.S.A. CALIFORNIA: Nash 38690, 38902, 38720 , 39279 (ASU), Bratt 12006, 12075 (SBBG), Tucker 33692b, 38658d (SBBG); Sheard 5054b-B, 5056a (SASK); UTAH: Wetmore 17095, 73625 (MIN). MEXICO. BAJA CALIFORNIA: Nash 38363 (ASU). ITALY. LIGURIA: Cevasco s.n. (M-0061350). DISCUSSION The general habit of the three species presented in this publication is extremely similar, and the species are therefore easily confused. It is not unusual to detect large amounts of calcium oxalates in the thallus of lichens from limestone substrates. calcium oxalates are common in epilithic lichens (Syers et al. 1967; Salvadori & Zitelli 1981; Ascaso et al. 1982; Jones & Wilson 1985; Wadsten & Moberg 1985; Bjelland et al. 2002). All species usually grow on calcium-rich substrate 190 that reacts with 10% HCl confirming the presence of calcium carbonate. The large amounts of calcium oxalates are not only visible in the polarized light microscope but also result in a strongly pruinose appearance of both the thallus and the apothecia. In some specimens pruina are, however, absent from the disk of the fruiting bodies. Without close examination, the rimose, heavily pruinose thalli both B. amabilis and B. subalbula could easily be mistaken for B. venusta (Figs. 21A–C, 22A,B, 23A,B). Indeed, one reason that B. subalbula has so far not been reported from North America is its close resemblance with B. venusta. Along the Californian coast both species grow closely together in the same habitat, sometimes side by side on the same substrate (Fig. 23B). Even though B. subalbula has slightly smaller apothecia, only microscopic examination will confirm the correct identification beyond any doubt. The most important characters to distinguish the three species are summarized in table 4, and a brief discussion of these characters follows. The Ascospores.—Buellia subalbula is generally characterized by oblong, one-septate spores (Fig. 21D,I). From more than six hundred spores examined, none of the spores had additional septa. In contrast, only young ascospores of B. venusta are one-septate, and additional septa develop as soon as the spores become pigmented (Fig. 23G). In general spores of B. venusta are not only larger than those of B. subalbula, but they also have a broader, more ellipsoid shape. Ascospores of B. amabilis are consistently one-septate (Fig. 22D) and generally larger than the ones of B. subalbula. In the TEM the perispore of both B. venusta and B. amabilis is considerably thicker than the proper spore wall. Mature spores of B. venusta are characterized by a finely 191 fissured perispore (Fig. 23I). At 1000x magnification a fine ornamentation is barely visible in the light microscope. Buellia amabilis shows strongly ornamented spores even at magnification as low as 400x (Fig. 22D). In the TEM the perispore is strongly fractured (Fig. 22E,F). In contrast B. subalbula has a thin, intact perispore in the TEM (Fig. 21I,K). In the light microscope the spore surface appears to be smooth and no ornamentation can be detected even with DIC. The exciple.—The differences in spore ornamentation and ultrastructure correlates with an absence of cinereorufa-green in both B. amabilis and B. venusta. Buellia subalbula is the only species with cinereorufa-green present in both exciple and epihymenium. The concentration of this diffuse pigment varies considerably but can be confirmed by a distinct HNO3+ violet reaction. In some specimens (especially the type material) the pigmentation appears strongly concentrated and deep bluish green. More typical is a diffuse dull brownish green pigmentation. Cinereorufa-green is absent in two specimens from Yemen (hb. Schultz 14151f, 14015e), which were therefore not included in B. subalbula s.str. Cinereorufa-green is a common pigment in the exciple and epihymenium of Buellia species with one-septate ascospores, but rarely encountered in species with pluriseptate spores (Nordin 2000)). The pigment is characteristic of the aethalea-type exciple sensu Scheidegger (1993). A strong carbonization of the outer part with cinereorufa-green and almost no structural differentiation of hyphae throughout is most characteristic for this exciple type. Apothecia often remain immersed, but in some species also emerge from the thallus. According to Scheidegger (1993), the aethalea-type can thus be observed in 192 cryptolecanorine, lecideine and zeorine apothecia. To clarify his concept, it should be emphasized that none of the apothecia in Buellia have a true lecanorine margin, i.e. an outer layer of thalline hyphae differentiated in cortex and medulla. The terms cryptolecanorine or zeorine may thus be misleading. These apothecia are not lecanorine and should more clearly be referred to as immersed lecideine apothecia. Apothecia of all three species can have a thalline collar, an irregular rim of loosely aggregated thalline material, which rarely remains attached to the outside of the lecideine exciple when the apothecium emerges from the thallus (visible e.g. in Fig. 21B). Internally this thalline collar appears not clearly differentiated from surrounding thallus material and should not be referred to as a thalline margin. The rimose thallus of all three species is little differentiated and cracks in the surface appear to be caused by irregular growth rather than distinct thallus differentiation. Thus, a thalline collar also appears to be a result of irregular growth. When Flotow (1850) first described the thalline collar as characteristic for Diplotomma, he was certainly not aware that this structure is rather ephemeral and not regularly observed in all specimens. Even in the same specimen, some apothecia may have a thalline collar whereas it is absent in others. Only the exciple of B. subalbula (Fig. 21F,G) can clearly be assigned to the aethaleatype sensu Scheidegger (1993). Apothecia of this species soon become sessile, but the exciple remains thin and poorly differentiated within. The inner hyphae extend radially from a deep reddish brown hypothecium. Hyphae of the hypothecium are strongly interwoven and can thus be characterized as textura intricata. Towards the exciple these hyphae become less pigmented and parallel in orientation. Both inner and outer excipular 193 hyphae are thin-walled (mesodermatous) and largely parallel (textura oblita). The inner hyphae are similar to the unpigmented paraphyses, whereas the cells of the outer hyphae are broadened and strongly pigmented (carbonized throughout). Although the anatomy of the exciple in B. venusta (Fig. 23C) is generally similar to that of B. subalbula, the strong carbonization of the outer hyphae is lacking. Only the outermost cells of the exciple are slightly swollen and like the paraphyses have a brown pigment cap. Unlike in B. subalbula, there is no distinctly carbonized outer exciple in both B. amabilis and B. venusta. The inner hyphae of the exciple in B. amabilis (Fig. 22C) are similar in color and orientation to the pale reddish brown hypothecium. These inner hyphae are more strongly interwoven than in B. subalbula or B. venusta (textura intricata) but become parallel towards the outside (textura oblita). The presence of intricately interwoven hyphae is characteristic for the leptocline-type sensu Scheidegger (1993), but the pale brown pigmentation of the hypothecium is quite unusual. In Rinodina the hypothecium typically is colorless. In the specimen of B. amabilis pigmentation is weak but nevertheless present. From these observations it is evident that the exciple types described by Scheidegger (1993) cannot easily be applied to all the material examined in this study. Buellia venusta has a structurally similar exciple without the characteristic outer carbonization and, although the exciple of B. amabilis resembles the leptocline-type, the hyphae are not as densely interwoven as in the exciple of B. leptocline or B. halonia, two other species assigned to this type according to Scheidegger & Ruef (1988) and Scheidegger (1993). 194 Therefore the exciple types described by Scheidegger (1993) may not be as strictly delimited as suggested. The structural arrangement of hyphae as well as the pigmentation are most certainly independent characters, and therefore not necessarily correlated. Within one species both pigmentation and anatomy are nevertheless very consistent and thus diagnostic. Other characters.—Ascus structure has been emphasized in the distinction between Buellia and Rinodina. In Lugol's iodine the tholus of Buellia shows two iodine blue flanks, which join at the ascus apex. The central part of the tholus generally does not react with iodine and appears either tapered (Bacidia-type) or with ±parallel flanks (Biatora-type). In contrast, in Rinodina the iodine blue flanks do not join at the tip but the central unstained area widens towards the ascus apex. This type is generally referred to as the Lecanora-type. All species treated in this study have an ascus type typical for Buellia (Fig. 21D). The distinction between Bacidia- and Biatora-type is not well pronounced, because there is variation in the shape of the central, iodine negative area of the tholus. Pycnidia were only found in B. subalbula and B. venusta. Both species have unilocular, globose pycnidia lined with short, moderately branched conidiophores (Fig. 23D). As described by Vobis (1980) for the type V, some conidia are formed laterally on bayonet-like projections. The conidiophores are, however, scarcely branched and relatively short (Figs. 21H, 23E,F). In addition, most conidia are formed from terminal cell projections. Buellia subalbula has short, bacilliform conidia (Fig. 21H). The conidia of B. venusta are at least twice as long, sometimes slightly tapered at both ends and some may thus appear ±fusiform (Fig. 23F). Scheidegger (1993)emphasized differences in 195 conidiophores and conidia when he recognized Amandinea as a genus separate from Buellia. Conidiophores and conidia of the “Diplotomma-group” are apparently not distinctly different from Buellia s. str. CONCLUSIONS The thin perispore, the presence of cinereorufa-green in the exciple and the absence of additional spore septa clearly exclude B. subalbula from the “Diplotomma-group” sensu Nordin (2000). Buellia amabilis may, however, be closely related to the “Diplotommagroup” even though it has one-septate instead of pluriseptate spores. Compared with B. venusta, the strongly thickened perispore supports the inclusion within Diplotomma. However, the perispore is much more fractured, and as a result a coarse rugulate ornamentation is distinct, even at moderate magnifications. In comparison with B. venusta structural differences in apothecial anatomy appear not very pronounced and even though the pigmentation of the B. amabilis exciple is not as strong, the pigments seem to be identical with the ones present in B. venusta. Consequently, our studies suggest that the genus concept of Diplotomma is still not clearly defined. Even if a more narrow concept sensu Nordin (2000) is adopted, some species with one-septate spores may eventually have to be included. ACKNOWLEDGEMENTS We are grateful to the curators of the herbaria cited for specimen loans. Bill Sharp and Dr. Robert Roberson (Arizona State University, ASU, U.S.A.), and Dr. Rosemarie Honegger (Zürich, Switzerland) provided valuable advice on scanning and transmission 196 electron microscopy. Dr. Laurence Garvie (ASU) kindly instructed the first author in powder x-ray diffraction. Scott Bates (ASU), Dr. John Sheard (Saskatchewan, Canada), Dr. Helmut Mayrhofer and Ulrike Grube (Graz, Austria) kindly reviewed the manuscript. This study was supported by a Research Grant in Plant Systematics from the International Association of Plant Taxonomists (IAPT) and two National Science Foundation grants (DEB-0103738, DEB-9701111). LITERATURE CITED ABU-ZINADA, A. H., D. L. HAWKSWORTH & H. A. BOKHARY. 1986. The lichens of Saudi Arabia, with a key to the species reported. Arab Gulf Journal of Scientific Research, Special Publication 2: 1-54. ASCASO, C., J. GALVAN & C. RODRIGUEZ-PASCAL. 1982. The weathering of calcareous rocks by lichens. Pedobiologia 24.: 219-229. BJELLAND, T., L. 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(Physciaceae, Lecanorales) in Europa. Nova Hedwigia 47: 433-468. SCHOLZ, P. 2000. Katalog der Flechten und flechtenbewohnenden Pilze Deutschlands. Schriftenreihe für Vegetationskunde 31: 1-298. SINGH, A. 1964. Lichens of India. Bull. Natl. Bot. Gard. [Lucknow] 93: 1-356. 200 SINGH, S. R. & D. D. AWASTHI. 1989. Lichen genus Diplotomma from India and Nepal. Geophytology 19: 173-181. SYERS, J., A. BIRNIE & B. MITCHELL. 1967. The calcium oxalate content of some lichens growing on limestone. Lichenologist 3: 409-414. SZATALA, O. 1955. Prodrome de la flore lichenologique de la Nouvelle Guinee. Ann. Hist.-Nat. Mus. Nationalis Hungarici (ser. nov.) 7: 15-50. THOMSON, J. W. 1997. American Arctic Lichens. 2. The Microlichens. University of Wisconsin Press, Madison. VOBIS, G. 1980. Bau und Entwicklung der Flechten-Pycnidien und ihrer Conidien. Bibliotheca Lichenologica 14: 1-141. VOBIS, G. & D. L. HAWKSWORTH. 1981. Conidial lichen-forming fungi, pp. 245-273. In COLE, G. T. & B. KENDRICK (eds.), 1: Biology of Conidial Fungi. Academic Press, New York. WADSTEN, T. & R. MOBERG. 1985. calcium oxalate hydrates on the surface of lichens. Lichenologist 17: 239-245. WHITE, F. J. & P. W. JAMES. 1985. New guide to microchemical techniques for the identification of lichen substances. British Lichen Society Bulletin 57: 1-41. WIRTH, V. 1994. Checkliste der Flechten und flechtenbewohnenden Pilze Deutschlands eine Arbeitshilfe [Checklist of the lichens and lichenicolous fungi of Germany]. Stuttgarter Beitrage zur Naturkunde, Serie A 517: 1-63. ———. 1995. Die Flechten Baden-Württembergs, Teil 1 & 2. Ulmer, Stuttgart. 201 PUBLICATION 5 (Canadian Journal of Botany, in press): Morphology and anatomy of chasmolithic versus epilithic growth: a taxonomic revision of inconspicuous saxicolous Buellia species from the Sonoran Desert Region generally ascribed to the “Buellia punctata”-group. F. BUNGARTZ, T. H. NASH III and B. D. RYAN School of Life Sciences, Arizona State University, PO Box 87 4601, Tempe, Arizona 85 287 - 4601, U.S.A. address correspondence to:Frank Bungartz, ph.: +1 480 965 7133, fax: +1 480 965 6899, e-mail: frank.bungartz@asu.edu. Abstract. Six saxicolous species of Buellia, which were previously generally identified as Buellia punctata, were examined from the Sonoran Desert. None of the species belongs to B. punctata s. str. Though inconspicuous, it can be demonstrated that the thallus morphology of these species is quite distinct and far less variable than previously assumed. Most species are epilithic even though their thalli also show some degree of substrate penetration. Buellia sequax, not previously reported from North America, is exclusively chasmolithic. Three new species with epilithic thalli, B. christophii, B. ryanii and B. tergua, are described. Two species with filiform conidia are not treated in the genus Amandinea. We discuss why the current delimitation of this genus, based solely on conidial shape, is rejected: Buellia pullata, with filiform conidia, is consequently not transferred into Amandinea. Amandinea. lecideina is synonymized with B. prospersa. 202 The taxonomy of crustose lichen species with thin and inconspicuous thalli is generally poorly resolved. These species are often overlooked, rarely collected and their thallus characteristics have been disregarded as highly variable and poorly differentiated. The attention of taxonomic treatments has therefore often focused on species with well developed epilithic thalli, whereas species which show some degree of endolithic growth have been neglected. This paper focuses on a group of species which in North America have generally been treated as Buellia punctata (Hoffm.) A. Massal., or more recently as Amandinea punctata (Hoffm.) Coppins & Scheid. Especially the saxicolous material commonly identified as this species is poorly understood (Mayrhofer & Moberg 2002). Thallus variation has largely been ignored. For example, in his treatment of the North American species of Buellia, Imshaug (1951) states: "… Buellia pullata is, as Tuckerman (1888) indicated, a saxicolous form of B. punctata with a well developed thallus. Buellia saxicola, on the other hand, is a saxicolous form of B. punctata with a scant to obsolete thallus …" The material examined in our study shows indeed some variation in thallus morphology. Several distinct species can nevertheless be distinguished based on thallus morphology as well as spore ontogeny, apothecial pigments, conidial length and secondary chemistry. Buellia pullata, for example, is not, as Imshaug (1951) suggested, a mere form of B. punctata, but a distinct species. Likewise, it can be demonstrated that B. saxicola is a synonym of B. sequax, a taxon not previously documented from North America, but recognized as a distinct species by Scheidegger (1993). 203 Six species previously treated in Imshaug's "B. punctata"-group, are distinguished among the Sonoran material. Thallus variation in this group is clearly much less pronounced than previously assumed. None of the species presented belongs to Buellia punctata s. str., which apparently does not occur on rock substrates in the Sonoran Desert Region. Contrary to recent practice, the genus Amandinea is not accepted. The taxonomy of that genus is currently not clearly resolved and all species are therefore treated as members of Buellia s. l. METHODS All specimens were examined with light microscopy using hand- and cryosections. Both conventional bright field microscopy (BF) as well as differential interference contrast (DIC) were used. Selected specimens were also studied with scanning electron (SEM) and transmission electron microscopy (TEM) according to a protocol described in detail by Bungartz et al. (2002). To improve dehydration and infiltration this protocol has been modified according to Bungartz and Nash (2004a). Representative specimens were cut with a slow speed diamond saw and the sections treated according to a protocol outlined by Bungartz et al. (2004a). To visualize hyphae penetrating dark stone substrates, the protocol was modified and specimens were stained with Lactophenol-Cotton-Blue (LCB) instead of Periodic Schiff Reagent (PAS). Sections were studied both with light and scanning electron microscopy as discussed in Bungartz et al. (2004a). Some of the sections were critically point dried and gold coated. All sections were studied in a JEOL JSM-840A SEM at accelerating voltages between 7 and 12 kV in secondary and backscattered emission mode. 204 Specimens were spot tested and routinely examined with standardized thin-layer chromatography (Culberson & Kristinsson 1970; Culberson & Johnson 1982; White & James 1985; Orange et al. 2001). To differentiate various xanthones, selected specimens were additionally examined with standardized High Performance Liquid Chromatography (HPLC, Feige et al. 1993). TLC-Plates were interpreted with the computer program WINTABOLITES (Mietzsch et al. 1994), and scanned for permanent record (Egan 2001). Spores measurements are given according to Nordin (2000). Pigment names follow Meyer and Printzen (2000). Figure plates were assembled and processed in Adobe Photoshop 7.0 and selective contrast adjustments were made to emphasize details like the apothecia in SEM micrographs. RESULTS Key to the species: 1 Thallus chasmolithic, discontinuous, usually of dispersed granules, growing among minerals of the substrate .......................................................................................... 2 Thallus epilithic, forming a thin, ± continuous crust on the substrate surface ......... 3 2(1) Thallus with a pale yellowish tinge, UV+ yellow to orange, with xanthones, premature ascospores ellipsoid, with distinct septum thickening, conidia filiform ... ................................................................................................................ B. prospersa Thallus pale brown to gray, UV–, without xanthones, premature ascospores narrowly oblong, without septum thickening, conidia bacilliform ............ B. sequax 205 3(1) Young apothecia erumpent, outer exciple in cross section carbonized by an aeruginose, HNO3+ violet pigment, conidia bacilliform ......................................... 4 Young apothecia emergent, outer exciple in cross section carbonized by a dull brown pigment, HNO3-, conidia bacilliform to filiform .......................................... 5 4(3) Thallus “leather” brown, rimose, not delimited by a distinct hypothallus, exciple deeply aeruginose, ascospores 10–15 x 6–9 µm, conidia 4–7 µm ............. B. tergua Thallus olive gray to brownish olive, distinctly areolate in the center, with an intermediate undifferentiated zone, delimited by a distinct black hypothallus, exciple dull fuscous brown, ascospores 9–13 x 4–8 µm, conidia 2–5 µm .. B. ryanii 5(3) Premature spores broad, almost globose, evenly thick walled, septum not thickened; exciple thick, outer carbonized zone usually >20 µm in cross section, conidia bacilliform .............................................................................. B. christophii Premature spores oblong to ellipsoid, not conspicuously broadened, with a thin spore wall, septum thin or thickened; exciple thin, outer carbonized zone usually < 20 µ in cross section, conidia filiform ..................................................................... 6 6(3) Thallus pale, usually with a yellow tinge, UV+ yellow to orange, with xanthones; spores with a distinct ± persistent septum thickening .......................... B. prospersa Thallus deep brown, rarely dull gray, UV-, without xanthones; spores with a brief stage of inconspicuous septum thickening ................................................ B. pullata 206 TABLE 5. Diagnostic differences of Buellia christophii, B. prospersa, B. pullata, B. ryanii, B. sequax and B. tergua thallus growth & color thallus morphology conidia exciple spores UV reaction & chemistry Buellia christophii epilithic; pale to deep brown rimose to rimose-areolate, rarely subsquamulose; without hypothallus bacilliform (short) thick, persistent, brown broad oblong (almost globose), equal wall thickening, early microrugulate UV negative, no xanthones, ± norstictic acid Buellia prospersa epilithic, rarely vaguely chasmolithic; pale ivory rimose to rimose-areolate; usually without a hypothallus, rarely thallus surrounded by black outline filiform (short to very long) thin, persistent, brown oblong-ellipsoid, ± persistent septum thickening, early microrugulate UV+ yellow to orange, usually with xanthones, ± norstictic acid, ± atranorin Buellia pullata epilithic; deep brown to gray rimose to rimose-areolate, (rarely subsquamulose); without hypothallus filiform (short to very long) thin, persistent, brown oblong-ellipsoid, brief septum thickening, early microrugulate UV negative, no substances detected Buellia ryanii epilithic; olive gray to brownish olive rimose to rimose-areolate (rarely subsquamulose); with distinct arachnoid black hypothallus bacilliform (short) thin, excluded, aeruginose (N+ violet) oblong-ellipsoid, no wall thickening, late microrugulate UV negative, no substances detected Buellia sequax chasmolithic, inconspicuous; pale brown to grayish granular to rimose; without hypothallus bacilliform (short) thin, persistent to excluded, brown narrow oblong, no wall thickening, late microrugulate UV negative, no substances or ± norstictic acid, rarely with arthothelin Buellia tergua epilithic; “leatherbrown” rimose to rimose-areolate; without hypothallus bacilliform (short) thin to moderately thick, persistent, aeruginose (N+ violet) oblong-ellipsoid, no wall thickening, late microrugulate UV negative, no substances detected 207 BUELLIA CHRISTOPHII Bungartz sp. nov. Thallus saxicolus, crustaceus, rimosus vel rimoso-areolatus, tenuis vel crassus, fulvus, sine hypothallo. Apothecia sessilia, lecideina, marginibus propriis crassis. Excipulum crassum, fulvum, sine pigmento aeruginoso, carbonaceum. Asci 8-spori. Sporae unisaeptae, late ellipsoideae vel globosae, 9–15 x 6–10 µm. Pycnidia globosa. Conidia bacilliformes, 2–4 x 1–1.5 µm. Thallus acida norstictica et connorstictica continens vel has substantias deficiens. TYPE: MEXICO. BAJA CALIFORNIA: Lighthouse area near Laguna Manuela; 28°15'00''N, 114°07'00''W; altitude: 100 m; on basalt; 21 February 1993; Nash 33979 (ASU–holotype designated here). Taxonomic note.—This species is named in honour of Dr. Christoph Scheidegger, Birmensdorf, Switzerland. Thallus (Figs. 25A,B and 29A-I) crustose, usually rimose to rimose-areolate, thin and ± continuous but distinctly epilithic, never chasmolithic, rarely becoming thickened and distinctly rimose-areolate to subsquamulose, not delimited by a hypothallus; surface matt to ± shiny, usually deep brown, rarely pale brown, usually smooth, but frequently roughened in areas where the surface appears eroded, epruinose, phenocorticate. Apothecia soon sessile, lecideine; proper margin conspicuously thickened, usually persistent, rarely excluded with age; disk black, epruinose, plane, rarely becoming slightly convex with age; exciple of aethalea-type (Fig. 25C,D) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura 208 → FIGURE 25. Buellia christophii (A-G) and Buellia pullata (H-N).—A. Overview of the epilithic rimose thallus (Nash 33979–holotype).—B. Close-up of a epilithic rimoseareolate thallus (Nash 32100).—C. Light micrograph of the athalea-type exciple (crosssection in water): the thick outer part consists of swollen cells, which are carbonized with the dull brown pigment cf. elachista-brown (Nash 33979–holotype).—D. SEM micrograph of an exciple cross-section (Nash 32100).—E. Light micrograph of broad premature ascospores with a conspicuous thickened wall (Nash 33979–holotype).—F. TEM micrograph of a premature ascospore: (1) thin, smooth perispore, (2) intermediate layer, (3) thick proper wall and (4) endospore (Nash 33979–holotype).—G. Light micrograph of a pycnidium with bacilliform conidia (Nash 33979–holotype). —H. Overview of the epilithic rimose-areolate thallus (Bolander 131–lectotype)—I. Close-up of the epilithic rimose-areolate thallus (Nash 41227).—J. Light micrograph of a thallus cross-section: (e) epinecral layer, (c) cortical layer, (a) algal layer (Nash 41227).—K. Light micrograph of an athalea-type exciple (cross-section in water): the thin outer part consists of swollen cells, which are carbonized with the dull brown pigment cf. elachistabrown (Nash 41227).—L. SEM micrograph of an exciple cross-section (Nash 11426).— M. Light micrograph of a pycnidium with filiform conidia (Nash 41227).—N. TEM micrograph of a mature ascospore: (1) thick, fractured perispore, (2) intermediate layer, (3) proper wall and (4) endospore (Nash 11426). 209 210 oblita), similar in structure and orientation to the paraphyses, often ± reduced and transient with the dull reddish brown hypothecium (leptoclinoides-brown, textura intricata); outer excipular hyphae parallel (textura oblita), cells moderately swollen and usually strongly carbonized with various amounts of brown pigments (cf. elachistabrown), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown). Asci 8-spored, clavate, Bacidia-type. Ascospores (Figs. 25E,F and 30) broadly ellipsoid to almost globose, never constricted, with obtuse ends, not curved, (9.0–)10.2–[11.7]–13.2(–15.0) x (6.0–)6.8–[7.7]–8.6(–10.0) µm (n = 81), one-septate; proper septum narrow, not thickening during spore ontogeny (Fig. 30), with septal pore canal, simple pore and undifferentiated pore plug; no ornamentation visible (in DIC); spore wall (Fig. 25F) differentiated into smooth, thin perispore (0.10– 0.20 µm), narrow intermediate layer (< 0.10 µm), thick proper spore wall (0.50–1.50 µm) and moderately thickened endospore (0.20–0.40 µm). Pycnidia (Fig. 25G) rare to common, globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogenous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980; Vobis & Hawksworth 1981); conidia (Fig. 25G) simple, bacilliform, 2.0–4.5 µm x 1.0–1.5 µm (n = 69) (Fig. 31). Chemistry.—No substances found or with norstictic and connorstictic acid (low concentrations only detected by TLC and / or HPLC). All spot tests negative (very rarely 211 forming orange needle-shaped crystals with K if observed with the compound scope). UV-. Thallus without amyloid reaction, only the apothecia amyloid in Lugol’s. Substrate and ecology.—All specimens examined are from low elevations, growing on siliceous mineral-poor coastal rock. The holotype is erroneously labeled as growing “on basalt”. The substrate is, however, a very fine grained andesite with intermediate to silicic acidity, poor in bases. Distribution (Fig. 7).—The species is currently known only from coastal southern California, U.S.A., Baja California and Baja California Sur, Mexico. Notes.—The species is superficially similar to B. pullata (Fig. 25H,G and 29A-I) and differences have been discussed in the note on that species. Table 5 gives an overview of the diagnostic characters available to distinguish all species discussed here. Material examined.—MEXICO. BAJA CALIFORNIA. Ryan 21513, Marsh 6198, Nash 34569 (ASU). BAJA CALIFORNIA SUR. Nash 29800 (ASU). U.S.A. CALIFORNIA. Los Angeles Co. Wetmore 73435 (MIN); Hasse Exs. 205 (CAS); Weber L-42660, L-42178, L42662, L- 42163 (COLO); Nash 32093, 32100, 32174a, Nash 32208, Nash 32141 (ASU). Mendocino Co. Tucker 35170 (SBBG). Monterey Co. Weber 8242, 8237 (COLO). San Luis Obispo Co. Tucker 36478 (SBBG); Nash 36982 (ASU). Santa Barbara Co. Printzen 49, 76 (hb. Printzen); Weber L-80178 (COLO); Nash 32386, 41174, 41181, Ryan 31359, Crayton s. n. (ASU). 212 → FIGURE 26. Buellia prospersa (A-E, all specimens: Wetmore 71896) and Buellia sequax (F-N).—A. Overview of the epilithic rimose-areolate thallus.—B. Close-up- of Fig. A.—C. Light micrograph of the athalea-type exciple (cross-section in water): the thin outer part consists of swollen cells, which are carbonized with the dull brown pigment cf. elachista-brown.—D. Light micrograph of mature ascospores.—E. Light micrograph of filiform conidia.—F. Chasmolithic granular thallus (Nash 33018).—G. Light micrograph of the athalea-type exciple (cross-section in water): the thin outer part consists of swollen, cells, which are carbonized with the dull brown pigment cf. elachista-brown (Nash 32187).—H. Light micrograph of a characteristically narrow, mature ascospore (Nash 32187).—I. SEM micrograph of an exciple cross-section (Nash 33018).—J. Light micrograph of conidiophores with bacilliform conidia (Nash 32187).— K. TEM micrograph of a Bacidia-type ascus tip (for designation of the different layers see Bellemère 1994): a- and b-layer are missing due to fixation artifacts, (c) outer electron opaque c-layer, (d1) d1-layer, i.e. the outer tholus, which has a distinctly laminated, fibrillar structure (in light microscopy this outer part stains deep blue with Lugol’s iodine), (d2) d2-layer, i.e. the inner tholus, which is not layered and ± homogeneous (not staining in Lugol’s iodine) and (oc) ocular chamber (Nash 32718).— L. TEM micrograph of a premature ascospore: (1) thin smooth perispore, (2) intermediate layer, (3) proper wall and (4) endospore (Nash 32718).—M. TEM micrograph of mature ascospore: (1) thick fractured perispore, (2) intermediate layer, (3) proper wall and (4) endospore. (Ryan 21689).—N. TEM micrograph of the spore septum (s) with a simple pore plug (p) (Nash 32718). 213 214 BUELLIA PROSPERSA (Nyl.) Riddle. Brookl. Bot. Gardens Memoirs 1: 114. 1918. Lecidea prospersa Nyl., Flora 63: 127. 1880. TYPE: U.S.A. VIRGIN ISLANDS: St. Thomas [original label data. S. Thomae Antillarum], 1878, D. Forel s.n. (H-NYL 9312– lectotype selected here!). Rinodina lecideina H. Mayrhofer & Poelt, Bibl. Lich. 12: 112 (1979). TYPE: IRELAND. KERRY. Dingle Peninsula. Near the village Ballyoughteragh, N Baille near Ballyferriter: on pasture walls [original label data: Eire/Irland: Co. Ciarrai / Kerry, Corca Dhuibhne / Dingle-peninsula, Umgebung des Weilers Ballyoughteragh N Baille an Fheirtearaigh / Ballyferriter, an Weidenmauern], August 1978, Poelt s. n. (GZU– holotype!). Amandinea lecideina (H. Mayrhofer & Poelt) Scheid. & H. Mayrhofer, in Scheidegger, Lichenologist 25: 342. 1993. Buellia punctata f. crassior (Erichsen) Zahlbr., Cat. Lich. Univ. 8: 591 (1932).— Buellia punctata f. crassior Erichsen, Das linke Untertraveufer: 151 (1932). TYPE: GERMANY, SCHLESWIG-HOLSTEIN. Lübeck, Coast of Dummersdorf, boulders along the beach below the village Stulperbank [original label data: Deutschland, SchleswigHolstein, Lübeck, Dummersdorfer Ufer, Strandblöcke unterhalb Stulperbank], April 1928, Erichsen s.n. (HBG–holotype). Buellia punctata f. litoralis (Erichsen) Zahlbr., Cat. Lich. Univ. 7: 397. 1931. - Buellia myriocarpa var. litoralis Erichsen, Verh. Bot. Ver. Provinz Brandenburg 72: 48 (1030). Type: GERMANY, SCHLESWIG-HOLSTEIN. Eastcoast of the Island Alsen, near Kettingholz, on boulders along the beach in the supralittoral zone [original label data: 215 Deutschland, Schleswig-Holstein, Insel Alsen, Ostküste bei Kettingholz, an Strandblöcken der supralitoralen Zone], July 1932, Erichsen s.n. (HBG–holotype). Taxonomic notes.—A specimen from MICH [rocks on a hill, Christiansted, St. Croix, March 17-25, 1923, collected by Britton & Kemp 79, determined by B. Fink] has been annotated by Fink as the type of “Buellia substigmata Fink”. This name has never been published by Fink and the specimen is not the type of Buellia substigmatea Müll. Arg [Proceed. Roy. Soc. Edinburgh 11:465 (1882)]. The MICH specimen has correctly been annotated by Imshaug as Buellia prospersa (Nyl.) Riddle. Buellia prospersa is the valid name for Rinodina (= Amandinea) lecideina, which is, however, not identical with Buellia lecidina Stein. ex Cohn [Kryptog. Flora von Schlesien 2(2) (1879)]. Even though the spelling of the species epithets “lecideina” and “lecidina” is almost identical, Buellia lecidina Stein. ex Cohn is a synonym of Rinodina occulta Körb. Thallus (Fig. 26A,B) crustose, rimose, moderately thin and ± continuous but distinctly developed and not chasmolithic, not delimited by a distinct hypothallus or with a black outline around the thallus; surface matt to ± shiny, usually pale ivory and smooth, rarely gray and slightly roughened (probably as a result of surface damage), epruinose, phenocorticate. Apothecia soon sessile, lecideine; proper margin excluded with age; disk black, epruinose, plane, becoming convex with age; exciple of aethalea-type (Fig. 26C) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), similar in structure and orientation to the 216 paraphyses, often ± reduced and transient with the dull reddish brown hypothecium (leptoclinoides-brown, textura intricata); outer excipular hyphae parallel (textura oblita), cells moderately swollen and usually strongly carbonized with various amounts of a brown pigment (cf. elachista-brown); pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed, paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown). Asci 8-spored, clavate, Bacidiatype. Ascospores (Figs. 26D and 30) oblong to ± ellipsoid, not constricted, with obtuse ends, not curved, (9.0–)12.0–[13.5]–15.0(–16.0) x (5.0–)6.2–[7.3]–8.3(–9.5) µm (n = 91), one-septate; proper septum soon with a ± persistent thickening, becoming reduced only in old spores (i.e. with a distinct Physconia-type or Orcularia-type ontogeny); ornamentation microrugulate (conspicuous in DIC at an early stage of the ontogeny). Pycnidia rare, globose, unilocular; at maturity almost entirely occupied by the long conidia and lined with short, scarcely branched conidiophores; conidiogenous cells terminal (conidiophore-type III sensu Vobis 1980); pycnidial ontogeny similar to the Roccella-type (sensu Vobis 1980; Vobis & Hawksworth 1981); conidia (Fig. 26E) simple, long bacilliform to filiform, 7.0–32.0 µm x <1.0 µm (n = 106) (Fig. 31). Chemistry.—No substances found, or with various amount of the following secondary metabolites: ± norstictic and connorstictic acid, ± atranorin and / or several xanthones (HPLC: 4,5-dichloro-3-O-methylnorlichexanthone, thuringione, 4,5dichlorolichexanthone, 2,4,5-trichlorolichexanthone). Spot tests usually negative, rarely K+ yellow to orange, C+ orange. UV+ bright yellow to orange. Thallus without amyloid reaction, only the apothecia are amyloid in Lugol’s. 217 Substrate and ecology.—Growing on siliceous mineral-poor coastal rock (generally HCl-). Distribution (Fig. 9).—Probably cosmopolitan but restricted to coastal areas. In the Sonoran Region presently known only from southern California, U.S.A., and Baja California, Baja California Sur, Sonora and Sinaloa, Mexico. Note.—In general B. prospersa can be easily recognized by a pale, distinctly epilithic thallus (Figs. 26A,B) and ascospores with a persistent median septum thickening (Figs. 26D and 30). Buellia pullata is microscopically similar, but has a deep brown to dark gray thallus (Figs 25H,I and 29L), which does not react with UV-light. No xanthones were detected in Buellia pullata. Although not previously reported, thalli of B. prospersa always react UV+ yellow to orange and xanthones are characteristic for this species. Poorly developed thalli of B. prospersa sometimes appear ± chasmolithic, and these specimens are then difficult to separate from forms of B. sequax with a well developed thallus (Fig. 28B). If filiform conidia (Fig. 26E) can be found, the material is easily recognized, because B. sequax has bacilliform conidia (Fig. 26J). In both species pycnidia are, however, quite rare. Young ascospores of B. sequax are typically distinctly narrowly oblong and not ornamented (Figs. 26H and 30). Only overmature and often disintegrating ascospores show a weak ornamentation, whereas a microrugulate ornamentation usually develops in young ascospores of B. prospersa (Fig. 30). Young ascospores of B. prospersa typically have a distinctly thickened median septum (Figs. 26D and 30), that can become very pronounced in some specimens. However, in some specimens it is difficult to find spores at this stage of the ontogeny. Median thickening of the spore 218 septum is absent from all stages of ascospore development in B. sequax (Fig. 30). In the Sonoran Desert Region, B. prospersa is restricted to coastal California, U.S.A. and Baja California, Mexico (Fig. 9). Buellia sequax is widely distributed throughout the Sonoran Desert (Fig. 10). Material examined.—MEXICO. BAJA CALIFORNIA. Scheidegger s. n. (hb. Scheidegger); Nash 33982 (ASU). BAJA CALIFORNIA SUR. Nash 12653, 12755 (ASU). JALISCO. Nash 20765 (ASU). SINALOA. Moberg 10267 (UPS); Wetmore 71896 (MIN); Nash 33680, 10088, 12248 (ASU); SONORA. Nash 25625, 12518, 12521, 10964 (ASU). ITALY. SARDINIA. Scheidegger Inv. Nr. 11097 (hb. Scheidegger). NEW ZEALAND. SOUTH ISLAND. Canterbury. Nash 19098 (ASU); Blaha 200 (GZU). Southland. Blaha 152 (GZU). U.S.A. CALIFORNIA. Santa Barbara Co. Nash 41312 (ASU). Ventura Co. Nash 38657 (ASU). BUELLIA PULLATA Tuck. Lich. Californ. p. 26. 1866. TYPE: U.SA. CALIFORNIA: Rocks on the coast. Bolander 131 (FH-3280–lectotype selected here!), Bolander 181, 150 (FH-3280–syntypes!). Taxonomic notes.—This species is not synonymous with B. punctata as suggested by Imshaug (1951). Several specimens from FH are mounted together on one large herbarium sheet with the FH accession number 3280. A type specimen is not designated in the protologue. From the available material we therefore select the specimen labeled Bolander (131) as the lectotype. Even though the other specimens are mounted on the same sheet and have the same FH accession number the specimens Bolander 181 and 150 219 cannot be regarded as isolectotypes because they have different collection numbers. Bolander 179, also mounted on the same sheet, has bacilliform conidia and it belongs to Buellia christophii. Thallus (Figs. 25H,I, 29J-O) crustose, usually rimose to rimose-areolate, thin and ± continuous but distinctly epilithic, never chasmolithic, rarely becoming thickened and distinctly rimose-areolate to subsquamulose, not delimited by a hypothallus; surface matt to ± shiny, usually deep brown, rarely pale brown, smooth, epruinose, phenocorticate. Apothecia soon sessile, lecideine; proper margin excluded with age; disk black, epruinose, initially plane, soon becoming convex; exciple of aethalea-type (Fig. 25K,L) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), similar in structure and orientation to the paraphyses, often ± reduced and transient with the dull reddish brown hypothecium (leptoclinoides-brown, textura intricata); outer excipular hyphae parallel (textura oblita), cells moderately swollen and usually strongly carbonized with various amounts of a brown pigment (cf. elachista-brown); pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown). Asci 8-spored, clavate, Bacidiatype. Ascospores (Figs. 25N and 30) oblong to ± ellipsoid, not constricted, with obtuse ends, not curved, (9.0–)11.1–[12.5]–13.8(–15.0) x (5.0–)6.2–[7.1]–8.0(–10.0) µm (n = 91), one-septate; proper septum soon thickening during spore ontogeny, becoming narrow with age (i.e. with an indistinct Physconia-type ontogeny), with septal pore canal, 220 simple pore and undifferentiated pore plug; ornamentation microrugulate (conspicuous in DIC at an early stage of the ontogeny); spore wall (Fig. 25N) differentiated into smooth to cracked, thin perispore (< 0.20 µm), narrow intermediate layer (< 0.10 µm), moderately thick proper spore wall (0.3–0.5 µm) and moderately thickened endospore (0.20–0.40 µm). Pycnidia (Fig. 25M) common, globose, unilocular; at maturity almost entirely occupied by densely branched conidiophores; conidiogenous cells mostly terminal, some intercalary (cf. conidiophore-type III sensu Vobis 1980); pycnidial ontogeny similar to the Roccella-type (sensu Vobis 1980; Vobis & Hawksworth 1981); conidia (Fig. 25M) simple, predominantly filiform, 6.0-43.0 µm x <1.0 µm (n = 156). Chemistry.—No substances found. All spot tests negative. Thallus without amyloid reaction, only the apothecia are amyloid in Lugol’s. Substrate and ecology: Growing on siliceous mineral-poor rock (generally HCl-). Distribution (Fig. 9).—Fairly common along the coast of southern California, U.S.A., Baja California and Baja California Sur, Mexico. One specimen has also been found further inland in Sonora, Mexico. Notes.—The thallus morphology (Figs. 25H,I and 29J-O) of this species is very similar to that of B. christophii (Fig. 25A and B, 29A-G). Pycnidia are relatively common in both species and if filiform conidia are found (Fig. 25M) the material is, thus, reliably distinguished from B. christophii, which has bacilliform conidia (Fig. 31). The two species can, however, also be clearly distinguished by other characters. Buellia christophii has lecideine apothecia with a thick, rather persistent margin (Fig. 25A,B,C). 221 Buellia pullata has a thin margin which relatively soon becomes excluded (Fig. 25H,I,J). Although the structure of the proper exciple is virtually identical (compare Fig. 25D with Fig. 25L), the layer of carbonized swollen outer cells is at least twice as thick in B. christophii (Fig. 25C). Young ascospores of B. pullata are oblong to ellipsoid (Fig. 30); in B. christophii young ascospores are rather broad, and often appear almost globose (Fig. 25E and 25F). Spores of B. christophii have no septum thickening (Fig. 30), whereas a median thickening of the spore septum can usually be observed at least in some of the younger spores of B. pullata (Fig. 30). Material examined.—MEXICO. BAJA CALIFORNIA. Scheidegger s. n. (hb. Scheidegger); Nash 38267, 38458, 38513b (ASU); Wetmore 75732 (MIN). SONORA: Wetmore 71644 (MIN). U.S.A. CALIFORNIA. Los Angeles Co. Weber L-42114 (COLO); Nash 32174 (ASU); Mendocino Co. Nash 25476, 11426, 11424b (ASU). Monterey Co. Nash 18896 (ASU). San Diego Co.: Bratt 8658 (SBBG). San Luis Obispo Co. Tucker 28829B (SBBG). Santa Barbara Co. Bratt 6394, Tucker 34719, Tucker 35711 (SBBG); Nash 32672, 41099, 41227, 41249, 41311, 33012 (ASU), Wetmore 73713 (MIN). Ventura Co.: Nash 37059, 37015 (ASU); Baltzo 7215 (UC). BUELLIA RYANII Bungartz sp. nov. Thallus saxicolus, crustaceus, areolatus vel subsquamulosus, tenuis vel crassus, olivaceus. Hypothallus atratus arachnoideus. Apothecia erumpentia vel sessilia, lecideina, marginibus propriis tenuibus. Excipulum tenue, fulvo-caeruleum, pigmentum aeruginosum continens, carbonaceum. Asci 8-spori. Sporae unisaeptae, ellipsoideae vel 222 oblongae, 9–13 x 4-8 µm. Pycnidia globosa. Conidia bacilliformia, 2-5 x 1-1.5 µm. Materiae chimicae nullae. TYPE: U.S.A. CALIFORNIA. Santa Barbara: Santa Cruz Island, 4.5 km E of radar station, ridge crest down N slope; 34°00'15''N, 119°37'30''W; altitude: 287 m; on schist in oak-pine woodland, 9 January 1994; Nash 32448b (ASU–holotype designated here!). Taxonomic note.—The species is named in memory of Dr. Bruce D. Ryan, the third author of this publication, who died from cancer before this article was printed. Thallus (Figs. 27A,B) crustose, areolate to subsquamulose, usually delimited by a distinct black arachnoid hypothallus; surface matt to ± shiny, deep olive gray to brownish olive, smooth, epruinose, phenocorticate. Apothecia initially immersed appearing aspicillioid, soon bursting through the thallus surface and becoming adnate to sessile, lecideine, rarely with remains of necrotic thalline material attached to the margin (thalline veil, Fig. 27B); proper margin ± persistent, rarely excluded with age; disk black, epruinose, plane, rarely becoming convex with age; exciple of aethalea-type (Fig. 27C) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), similar in structure and orientation to the paraphyses, often ± reduced and transient with the dull reddish brown hypothecium (leptoclinoides-brown, textura intricata); outer excipular hyphae parallel (textura oblita), cells moderately swollen and usually strongly carbonized with various amounts of brown and aeruginose (HNO3+ violet) pigments (cf. elachista-brown & cinereorufa-green); pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; 223 paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown) and a diffuse aeruginose pigment (HNO3+ violet, cinereorufagreen). Asci 8-spored, clavate, Bacidia-type. Ascospores (Fig. 27D,E) broadly oblong to ellipsoid, with obtuse ends, not curved, mature spores slightly constricted, (9.0–)10.5– [11.3]–12.2(–13.0) x (4.0–)5.6–[6.2]–6.8(–8.0) µm (n = 100), one-septate; proper septum narrow, not thickening during spore ontogeny; ornamentation absent in immature and premature spores, microrugulate in mature spores (best seen in DIC). Pycnidia (Fig. 27E) rare, globose, unilocular; at maturity almost entirely occupied by densely branched conidiophores; conidiogenous cells mostly terminal, but some also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicariatype (sensu Vobis 1980; Vobis & Hawksworth 1981); conidia (Fig. 27F) simple, bacilliform to ellipsoid, 2.0–5.0 µm x 1.0–1.5 µm (n = 73) (Fig. 31). Chemistry.—No substances found and all spot tests negative. Thallus without amyloid reaction, only the apothecia are amyloid in Lugol’s. Substrate and ecology: All specimens examined grow on mineral-poor rock substrates such as quarzite and schist in open oak-conifer forest. Distribution (Fig. 9).—The species is currently known only from Santa Cruz Island, along the coast of southern California, U.S.A. and from Isla de Guadalupe, off the coast of Baja California, Mexico. It has not been found close to the sea shore but in open, dry oak-conifer woodland. Notes.—Scheidegger (1993) mentions the presence of pigment A (= cinereorufagreen) as characteristic of the aethalea-type exciple. All species discussed here, have an 224 → FIGURE 27. Buellia ryanii (A-F). and Buellia tergua (G-L, all specimens: Nash 38368– holotype).—A. Overview of the epilithic areolate thallus delimited by a distinct, black arachnoid hypothallus (Nash 38368–holotype).—B. Close-up of Fig. A: young apothecia aspicillioid, emerging from the thallus partially covered by a thalline veil (arrows).—C. Light micrograph of the athalea-type exciple (cross-section in water): the thin dull olive outer part consists of swollen cells which are carbonized by the dull brown pigment cf. elachista-brown and the aeruginose pigment cinereorufa-green (Nash 38368– holotype).—D. Light micrograph of mature ascospores (Nash 38368–holotype).—E. Light micrograph of a pycnidium with bacilliform conidia (Nash 32448b).—F. Light micrograph of conidiophores with bacilliform conidia (Nash 32448b).—G. Rimoseareolate thallus.—H. Light micrograph of the athalea-type exciple (cross-section in water): the moderately thickened strongly aeruginose outer part consists of swollen cells, which are carbonized by the dull brown pigment cf. elachista-brown and the aeruginose pigment cinereorufa-green.—I. Light micrograph of an exciple (cross-section in HNO3): the aeruginose pigment reacts violet and is partially diluted, thus the structure of the hyphal cells becomes more distinct.—J. Premature (p) and mature (m) ascospore.—K. Light micrograph of a pycnidium with bacilliform conidia.—L. Light micrograph of conidiophores with bacilliform conidia. 225 226 exciple with the structure of the aethalea-type, but the pigment cinereorufa-green is only present in the exciple and epihymenium of B. ryanii and B. tergua. The two species are quite similar and currently only known from a few localities off the coast of Baja California, Mexico and southern California, U.S.A. Buellia ryanii has an areolate to subsquamulose, deep olive gray to brownish olive thallus with a distinct arachnoid hypothallus (Fig. 27A,B). Buellia tergua has a leather brown rimose-areolate thallus with no hypothallus (Fig. 27G). The apothecial ontogeny is very similar. Apothecia are initially immersed and the margin is covered by the surrounding thallus. Thus, these apothecia look aspicillioid (Fig. 27B). Soon apothecia erupt from the surrounding thallus and become adnate. Rarely, thallus material remains attached to the lecideine margin as a thalline veil. The pigment cinereorufa-green is strongly concentrated in the exciple and epihymenium of B. tergua and in cross-section clearly visible as deeply aeruginose. In contrast, the exciple and epihymenium of B. ryanii is dull olive-brown and the presence of cinereorufa-green may thus be overlooked if not tested with HNO3. The spores of the two species have a similar ontogeny and become constricted with maturity (Fig. 30). However, spores of Buellia tergua are distinctly larger (10–15 x 6–9 µm) than those of B. ryanii (9–13 x 4–8 µm). The ornamentation of Buellia tergua spores develops early; it becomes visible at the premature stage and is distinct at maturity (Fig. 30). In B. ryanii spore ornamentation is barely discernible in mature spores but well developed in overmature spores (Fig. 30). The conidia of B. ryanii are slightly smaller (2-5 µm) than those of B. tergua (4–7 µm) (Fig. 31). 227 Material examined.—U.S.A. CALIFORNIA. Santa Barbara Co. Tucker 35781A, Bratt 3488b (SBBG); Wetmore 74077 (MIN); Nash 32445, Ryan 31621 (ASU). BUELLIA SEQUAX (Nyl.) Zahlbr. Cat. Lich. Univ. 7: 410. 1931. Lecidea sequax Nyl., Flora, Jena 58: 302. 1875. TYPE: FRANCE, HAUTES ALPES: on quartzite along the banks of the river Vienne near Moulin de l’Aiguille [Quartz sur les coteaux de la Vienne près du Moulin de l’Aiguille], 11 January 1872, Lamy 1083 [H-NYL 9538–lectotype selected by Scheidegger (1993)], H-NYL 9539–isolectotype, M-0023804– isolectotype!]. Buellia abstracta (Nyl.) H. Olivier, Bull. Acad. Intren. Géogr. Bot. 12: 176. 1903.— Lecidea abstracta Nyl., Flora, Jena 66: 102 (1883). Type: FRANCE, PYRENNÉES. Cauterets, E. Lamy (H-NYL 9740–holotype, M-0023906–isotype!). B. punctatula Malme, Arkiv för Botanik 21a(14):9 & 14. 1927. Type: PARAGUAY. ASUNÇIÓN. "Zapitapunta", on sunny sandstone [original label data: "Zapitapunta", ad rupes arenarias sat apricas], 2 July 1893, Malme 1424 (S–lectotype selected here!). Buellia saxicola de Lesd., Ann. Crypt. Exot. 5: 127 (1932). Type: U.S.A. NEW MEXICO. San Miguel: around Las Vegas, Agua Zarca, on siliceous rocks [original label data: environs de Las Vegas, Agua Zarca, sur roches siliceuses], Arsène Brouard 1930 (W acquisition 1934 no. 726–lectotype selected here!; UPS!, S!–isolectotypes). Taxonomic notes: The type material of Buellia saxicola is not identical with B. punctata as suggested by Imshaug (1951). The lectotypification of the material designated by Bouly de Lesdain is necessary because it can generally be assumed that his 228 entire holotype collection, originally located in Dunkerque (France), has been destroyed during the Second World War. The lectotype from Vienna was selected because this is the only material which still has a few intact pycnidia. Mayrhofer and Moberg (2002) list Buellia myriocarpa (DC.) De Not. as a synonym of Buellia punctata (= Amandinea punctata). In 1903 Hasse determined his exsiccati specimen no. 204 as Buellia myriocarpa (Lamb & DC.) Mudd. The exsiccati specimen at ASU is identical with Buellia sequax. Type material of B. myriocarpa was, however, not examined. Thallus (Figs. 26E and 28A-N) crustose, thin and discontinuous, ± chasmolithic, i.e. forming inconspicuous, poorly delimited granules hidden among the mineral grains of the substrate; thicker thalli rarely becoming continuous in parts, rimose to very rarely rimoseareolate, not delimited by a hypothallus; surface matt, pale brown to greyish, smooth to slightly roughened, epruinose, phenocorticate. Apothecia soon becoming adnate to sessile; lecideine; proper margin soon excluded with age; disk black, epruinose, plane, becoming convex with age; exciple of aethalea-type (Figs. 26G,I) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), similar in structure and orientation to the paraphyses, often ± reduced and transient with the dull reddish brown hypothecium (leptoclinoides-brown, textura intricata); outer excipular hyphae parallel (textura oblita), cells moderately swollen and usually strongly carbonized with various amounts of brown pigment (cf. elachistabrown); pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed, paraphyses simple to moderately branched, apically swollen, with a brown 229 pigment cap (cf. elachista-brown). Asci 8-spored, clavate, Bacidia-type (Fig. 26K). Ascospores (Figs. 26H,L-N and 30) distinctly narrowly-oblong (Fig. 26H), becoming ellipsoid with age, not or rarely constricted with age, with obtuse ends, not curved, (10.0– )10.7–[11.7]–12.7(–14.0) x (3.0–)3.8–[4.6]–5.4(–6.0) µm (n = 50), one-septate, proper septum narrow, not thickened during spore ontogeny, with septal pore canal, simple pore and undifferentiated pore plug (Fig. 26N); ornamentation absent from the early spore ontogeny (Fig. 26L), becoming visible in mature spores (Fig. 26M); spore wall differentiated into smooth to fissured, thin perispore (0.1–0.2 µm), narrow intermediate layer (< 0.1 µm), thick proper spore wall (0.3–0.5 µm) and moderately thickened endospore (0.2–0.3 µm). Pycnidia rare, globose, unilocular; at maturity almost entirely occupied by densely branched conidiophores; conidiogenous cells intercalary and terminal (conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980; Vobis & Hawksworth 1981); conidia (Fig. 26J) simple, bacilliform, 2.0–4.0 µm x 1.0–1.5 µm (n = 50) (Fig. 31). Chemistry.—No substances found or with norstictic acid and / or arthothelin (low concentrations only detected by TLC and / or HPLC). All spot tests negative (very rarely forming orange needle-shaped crystals with K if observed with the compound scope). UV-. Thallus without amyloid reaction, only the apothecia amyloid in Lugol’s. Substrate and ecology: Growing on a large variety of siliceous rocks (generally HCl-). Distribution (Fig. 10).—The species is widely distributed throughout the North American Southwest. In the Sonoran Desert Region, specimens have been found from coastal up to subalpine localities. 230 → FIGURE 28. Chasmolithic growth in Buellia sequax on various substrates.—A. Light micrograph of a thallus on a sandstone, almost entirely composed of quartz (substrate crosssection): the poorly delimited thallus is growing between the quartz crystals (Nash 33018).—B. Light micrograph showing the thallus aspect of well developed cortical granules (arrows) between feldspar crystals (Nash 32718).—C. Light micrograph of a thallus with corticate granules growing between feldspar crystals (substrate cross-section): arrows point at the algal layer (Nash 32718).—D. SEM micrograph of two apothecia and poorly delimited hyphae growing between quartz crystals of a sandstone (specimen critically point dried, gold coated; Nash 33018).—E. Light micrograph of a chasmolithic thallus almost entirely hidden in the fissures of a foliated metamorphic low-grade phyllite (Weber L-42776).—F. Light micrograph of a thallus on phyllite (substrate cross-section): the phyllite contains some quartz crystals, and it is rich in mica and thus ± foliated; hyphae have been stained with lactophenol-cotton-blue, which results in the darkening of the upper, inhabited layers of the substrate (Weber L-42776).—G. Natural break of a sandstone where the fractures between the quartz crystals are inhabited by the thallus (Nash 40151).—H. SEM micrograph of the chasmolithic thallus on phyllite (critically point dried, gold coated): the minerals appear somewhat coated but no distinct hyphae are visible (Weber L-42776).—I. Cross-section of the specimen growing on phyllite (critically point dried, gold coated): the thallus hyphae (t) are closely conglutinated between the mineral crystals (c) of the substrate (Weber L-42776).—J. SEM micrograph of a thallus growing on sandstone (critically point dried, gold coated): the thallus consists of hyphae and algal cells growing irregularly between the quartz crystals (Ryan 21689).—K. Detail of Fig. J: thallus hyphae (t) growing between quartz crystals (c). L. Detail of Fig. K: the haustorial hyphae (h) are closely wrapped around algal cells (a), these thallus areas are developed between the quartz crystals (c) of the substrate.—M. SEM of a chasmolithic thallus on sandstone (cross-section resin-embedded, polished and carbon coated): (a) apothecium, (t) thallus, (c) quartz crystal (Ryan 21689).—N. SEM of a chasmolithic thallus on a ± metamorpic volcanic rock with fractured and sheared mineral crystals (resin-embedded cross-section, polished and carbon coated): (s) stipe of the apothecium, (e) exciple, (t) thallus, (c) sheared mineral crystals (Nash 32187). 231 232 Notes.—Specimens of B. sequax with an unusually well developed thallus are sometimes superficially similar to B. prospersa and diagnostic differences have been discussed under that species. Specimens examined.—AUSTRIA. TIROL. Arnold s. n. (M-0061303). MEXICO. BAJA CALIFORNIA. Scheidegger s. n. (hb. Scheidegger), Nash 26137, 38513a, 40151, 26336, 34608 (ASU). BAJA CALIFORNIA SUR. Scheidegger s. n. (hb. Scheidegger); van den Boom 25063 (hb. van den Boom); Nash 33718, 33858, 40071, 40101, 26144 (ASU) SONORA: Ryan 21689 (ASU). ITALY. ELBA. Hertel 6723 (M-0061306), Triebel & Rambold 6209 (M-0061304), Albertshofer s. n. (M-0061303). SICILY. Doppelbaur s.n. (M-0061307). SPAIN. MURCIA. Doppelbaur s.n. (M-0061305). U.S.A. ARIZONA. Apache Co. Ryan 19184 (ASU) Cochise Co. Weber S-8754 (COLO); Darrow 1970 (ASU). Coconino Co. Nash 35171, Boykin 2725 (ASU). Gila Co. Nash 28487, Schramm 225 (ASU). Maricopa Co. Scheidegger s. n. (hb. Scheidegger); Nash 9605a (ASU). Pinal Co. Scheidegger s. n. (hb. Scheidegger). CALIFORNIA. Los Angeles Co. Wetmore 73249 (MIN); Hasse L- 78819 (COLO); Weber L-42778, L- 42776 (COLO); Bratt 10196 (SBBG); Nash 32187, Hasse Exs. 204, Ryan 30929b (ASU); Hasse Exs. 53 (CAS). Merced Co. Tucker 28820 (SBBG), Tucker 28815 (ASU). Monterey Co. Nash 8035 (ASU). Orange Co. Weber L-42052 (COLO). San Diego Co. van den Boom 25192 (hb. van den Boom); Bratt 3592 (ASU). San Luis Obispo Co. Tucker 28829A, 36471 (SBBG); Tavares 1409 (UC). Santa Barbara Co. Tucker 35689A, 35845, 35781, 36463, 35781, Bratt 11388, 5709, 3641, 3643, 10316, 3488a (SBBG); Printzen 74, 76 (hb. Printzen), Nash 32352, 32656, 32747, 32837, 41392, 41423, 33016, 33018, 32890; Bratt 3488, 7681, 8879A, Ryan 31216 (ASU). Ventura Co. Bratt 233 10392, Hamber 7 (SBBG) NEW MEXICO. San Juan Co. Nash 16225, 16396 (ASU). Valencia Co. Nash 16004 (ASU). UTAH. Washington Co. Nash 15316 (ASU). WYOMING. Platte Co. Ryan 14306 (ASU). BUELLIA TERGUA Bungartz sp. nov. Thallus saxicolus, crustaceus, rimosus vel rimoso-areolatus, tenuis vel crassus, alutaceus, sine hypothallo. Apothecia erumpentia vel sessilia, lecideina, marginibus propriis tenuibus. Excipulum tenue, aeruginosum, pigmentum aeruginosum continens, carbonaceum. Asci 8-spori. Sporae unisaeptae, ellipsoideae vel oblongae, 10–15 x 6–9 µm. Pycnidia globosa. Conidia bacilliformia, 4–7 x 1–1.5 µm. Materiae chimicae nullae. TYPE: MEXICO. BAJA CALIFORNIA, Isla de Guadalupe: SE of southern peak; 28°57'30''N, 118°15'00''W; altitude 600 m; on basalt, shaded exposure within canyon with rocky outcrops and some native perennials; 3 January 1996, Nash 38368 (ASU– holotype designated here!). Taxonomic note.—The species name is derived from the Latin word for leather, tergus. Thallus (Fig. 27F) crustose, rimose to rimose-areolate, not delimited by a distinct hypothallus; surface matt, usually deep brown and smooth, epruinose; phenocorticate. Apothecia initially immersed appearing aspicillioid, soon bursting through the thallus surface and becoming adnate to sessile, lecideine, rarely with remains of necrotic thalline material attached to the margin (thalline veil); proper margin ± persistent, rarely excluded 234 → FIGURE 29. Variation of epilithic growth in Buellia christophii (A-I; Nash 32100, on silica-rich rhyolithe, Nash 33979–holotype, on a vesicular, fine-grained volcanic andesite) and B. pullata (J-O; Nash 32672, on a fine-grained sandstone with ± metamorphic, laminated and flattened grains).—A. Light micrograph showing the rimose-areolate epilithic thallus aspect on rhyolithe (Nash 32100).—B. Light micrograph of a thallus on rhyolite (substrate cross-section): the thallus is distinctly developed on the surface (Nash 32100).—C. Detail of Fig. B: a single areole with an apothecium.—D. SEM micrograph of a thallus on andesite (substrate cross-section critically point dried, gold coated): the thin thallus is distinctly developed on the surface (Nash 33979– holotype).—E. Detail of Fig. D: the hyphae are restricted to the substrate surface. 60. Backscattered SEM micrograph (resin-embedded, polished, carbon coated): (a) apothecium, (t) epilithic thallus and (h) hyphae penetrating natural crevices (Nash 32100). 61. Detail of Fig. F: (t) thallus, (c) substrate crevice penetrated by a bundle of hyphae, the arrow points to a narrow fissure.—E. SEM micrograph of a thallus on andesite (substrate cross-section resin-embedded, polished, carbon coated): (a) apothecium, (t) thallus, (s) substrate (Nash 33979 - holotype).—F. Detail of Fig. E: (c) thallus cortex, (a) alga, (q) quartz crystal.—F. SEM micrograph of the rimose-areolate thallus on metamorphic sandstone (specimen critically point dried, gold coated).—G. Detail of Fig. F.—H. Light micrograph of the rimose-areolate thallus.—I. Light micrograph of a thallus on sandstone (substrate cross-section): the thallus areoles are distinctly developed on the surface.—J. SEM micrograph of a thallus on sandstone (substrate cross-section critically point dried, gold-coated): the thallus areoles are distinctly developed on the substrate surface; (a) apothecium, (t) thallus, (c) natural cavity penetrated by lichen hyphae.—K. SEM micrograph of a thallus on sandstone (substrate cross-section polished resin-embedded specimen, carbon coated): (t) thallus, (s) substrate. 235 236 → FIGURE 30. Ascospore ontogeny of Buellia species with inconspicuous saxicolous thalli from the Sonoran Desert: Immature ascospores are hyaline, mostly non-septate (Bontogeny), but some are early septate (A-ontogeny). An olive pigmentation begins to develop at the premature stage, some spores also showing wall and septum-thickening. The mature spores become brown, the ornamentation – if present – is barely visible at this stage and wall and septum thickenings are becoming reduced. Overmature spores are deep brown and the ornamentation – if present – is distinctly developed; wall and septum thickenings are largely reduced at this stage. 237 238 with age; disk black, epruinose, plane, rarely becoming convex with age; exciple of aethalea-type (Fig. 27H,I) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), similar in structure and orientation to the paraphyses, often ± reduced and transient with the dull reddish brown hypothecium (leptoclinoides-brown, textura intricata); outer excipular hyphae parallel (textura oblita), cells moderately swollen and usually strongly carbonized with various amounts of brown and aeruginose (HNO3+ violet) pigments (cf. elachista-brown & cinereorufa-green); pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown) and a diffuse aeruginose pigment (HNO3+ violet, cinereorufagreen). Asci 8-spored, clavate, Biatora-type. Ascospores (Figs. 27J and 30) broadly oblong to ellipsoid, with obtuse ends, not curved, mature spores slightly constricted, (10.0-)11.1-[12.2]-13.4(-15.0) x (6.0-)6.0-[6.6]-7.3(-9.0) µm (n = 68), one-septate; proper septum narrow, not thickening during spore ontogeny (i.e. Buellia-type); ornamentation absent in immature and premature spores, microrugulate in mature spores (best seen in DIC). Pycnidia (Fig. 27K) rare, globose, unilocular; at maturity almost entirely occupied by densely branched conidiophores; conidiogenous cells intercalary and terminal (conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicariatype sensu Vobis 1980; Vobis & Hawksworth 1981); conidia (Fig. 27L) simple, bacilliform, 4.0-7.0 µm x 1.0-1.5 µm (n = 50). Chemistry: No substances found and all spot tests negative. Thallus without amyloid reaction, only the apothecia are amyloid in Lugol’s. 239 Substrate and ecology.—On siliceous volcanic coastal rock. Distribution (Fig. 11).—Presently known only from Isla Guadalupe, off the coast of Baja California, and two localities on the Pacific coast of mainland Baja California, Mexico. Notes.—The thallus of B. tergua is very similar to B. pullata (Fig. 25H,I) and B. christophii (Fig. 25A,B), even though these two species are generally darker brown. Apothecia of B. tergua, erupt from the thallus and young apothecia thus appear aspicillioid. In B. pullata and B. christophii, apothecia emerge more gradually and young apothecia appear immersed but not aspicillioid. Buellia ryanii is the only other species with cinereorufa-green in the apothecia and diagnostic differences are presented in the notes on that species. Material examined.—MEXICO. BAJA CALIFORNIA. Moberg 8588 (UPS); Nash 38366a, 38368, 8736, 38442 (ASU). DISCUSSION Thallus Variation: chasmolithic vs. epilithic growth.—All species discussed here, are often described as having a scant or obsolete thallus and were previously interpreted as poorly developed forms of B. punctata rather than assigned independent species rank. However, in the Sonoran Desert the saxicolous species of the "Buellia punctata"-group show considerable diversity. The thallus morphology of the species is quite diagnostic and far less variable than commonly implied. The extent to which a thallus is developed on the surface or partially hidden within crevices, pores or fissures is necessarily related to substrate composition and structure. 240 → FIGURE 31. Range of conidial length in various saxicolous species of Buellia (modified from Scheidegger 1993). The bars indicate the range of measurements for each of the species. Buellia pullata overall has not only the longest conidia but also the largest variation in measurements. For B. prospersa the bold bar indicates the range of measurements given by Scheidegger (1993), the narrow bar indicates the variation observed here. Measurements overlap considerably and the asymptotic graph shows that the distinction of bacilliform vs. filiform is arbitrary and does not represent a discrete character. 241 242 Both epilithic and chasmolithic thalli clearly show some degree of substrate penetration, provided that enough space is available (Figs. 28 and 29). The species examined here, all grow on mineral-poor weathering rinds of various siliceous rocks. Although the composition of these substrates varies considerably, they are all poor in nutrients such as iron, magnesium or calcium, and no calcium carbonates were found. The fungal hyphae are apparently not able to dissolve their substrate and instead, hyphal penetration is confined to the pre-existing crevices (e. g., Figs. 28M,N and 29F,G). Both epilithic and chasmolithic thalli can thus be distinguished from truly endolithic lichens, which develop entirely within their substrate and are at least partially able to dissolve some of the minerals Bungartz et al. (2004a). Generally, chasmolithic growth (Fig. 28A) and epilithic growth can be distinguished (Fig. 29): Epilithic thalli are well developed on the substrate surface, internally distinctly stratified (Figs. 25J and 29I) and have a ± continuous surface (Fig. 29D,E,J,K). Although these thalli may be thin and inapparent, they are nevertheless largely covering the substrate (Figs. 25A,B,H,I, and 29A,L). In contrast, chasmolithic thalli grow poorly differentiated among the mineral grains (Fig. 29A,D,G,J), these thalli are less stratified and sometimes only consist of loosely associated hyphae (Fig. 28J-L). Even well developed chasmolithic thalli (Fig. 28B,C) do not establish a continuous surface crust. Among the species examined here, only B. sequax may consistently be referred to as chasmolithic, i.e. the major part of the thallus remains hidden among the mineral grains, even though some areas may develop into a discontinuous crust (Fig. 28G). Specimens of B. sequax with well developed thallus areas can be distinguished from the other, epilithic 243 species by the pale thallus colour. Only poorly developed or damaged thalli of B. prospersa may be confused with these well developed specimens of B. sequax. Comparing the types of B. abstracta and B. sequax Scheidegger (1993) argued that B. abstracta s. str. with narrow ascospores and a chasmolitic thallus cannot reasonably be distinguished from epilithic and often areolate material assigned to B. sequax s. str. He therefore synonymized B. abstracta with B. sequax, which he regarded as a highly polymorphic species. We agree that both taxa can be synonymized. Buellia sequax is, however, far less variable than this statement may imply. Chasmolithic growth is not merely the result of substrate variation because Buellia sequax consistently forms chasmolithic thalli on a variety of different substrates. We have examined type material of the two taxa as well as material from M annotated by Scheidegger. Buellia sequax s. str. indeed shows ± larger and better developed thallus areas, but even well developed thalli remain poorly delimited patches and never spread extensively into a continuous crust. Among the species discussed here, only B. ryanii forms a distinctly areolate thallus (Fig. 27A,B). The areoles form independently on a black, arachnoid hypothallus, and individual areoles regularly become subsquamulose along the margin. All other species have a rimose to rimose-areolate thallus, i.e. the surface is initially fissured and only secondarily breaks into areoles. In exceptionally thick thalli of B. christophii and B. pullata these secondary areoles can even become subsquamulose. Generally, thalli of B. christophii, B. pullata and B. tergua are very similar and easily confused (Figs. 25A,B, H,I and 27G). Buellia tergua is lighter brown than the deep brown thalli of the other two 244 species. However, only a few specimens of B. tergua are currently known and it is therefore not possible to assess whether the leather-brown colour represents a consistent trait. The three species may therefore not reliably be distinguished by their thalli alone. Spore Ontogeny and Ultrastructure.—In crustose Physciaceae, spore structure has attracted considerable attention (Mayrhofer 1982) and, especially in the genus Rinodina, several diagnostic types have been recognized because of conspicuous wall and septum thickenings (Mayrhofer & Poelt 1979; Mayrhofer 1984b; Giralt 2001). In Buellia Scheidegger (1993) first discovered thickened spore septa in a few species, and Kalb (1986) segregated the genus Hafellia for species with lateral wall thickenings. The wall thickenings of spores in the Physciaceae are, however, by no means static but the result of a dynamic spore ontogeny, which is still not completely understood. All species treated here have very similar ascospores and measurements overlap considerably. Nevertheless, diagnostic differences can be found in all species by carefully comparing their spore ontogeny (Fig. 30). For convenience, four stages may be distinguished and some characters are only present during a brief stage: Immature spores are hyaline, premature spores olive, mature spores brown and overmature spores deep brown. Immature and premature spores generally have no visible ornamentation, although in some species ornamentation may begin to develop with the onset of pigmentation. Other species barely show any ornamentation during their ontogeny, but overmature spores often appear ornamented. This may sometimes be an artefact ascribed to degeneration of the spores. Wall or septum thickenings are usually best developed in premature and mature spores and may disappear again in aging spores. 245 In Rinodina Giralt and Mayrhofer (1994a; 1995) distinguish spores, that form spore septa before apical wall thickenings (A-ontogeny), from spores with late septum formation, where apical thickenings are present before the development of a septum (Bontogeny). Giralt (2001) admits that some species have an intermediate ontogeny. In spores typically found in Buellia, apical thickenings generally do not develop (Fig. 30). Early septum formation may better be distinguished from late septum formation based on spore wall pigmentation. In spores with A-ontogeny, pigmentation develops after a septum can be distinguished in the immature spore. Late septum formation of the Bontogeny develops in premature spores after the walls have become distinctly pigmented. Buellia tergua and B. sequax can tentatively be assigned to the A-ontogeny, whereas spores of the other species appear to follow the B-ontogeny. This distinction is not easy to assess. Even spores with pigmented walls do not always have septa, and sometimes these spores can also be observed along hyaline spores with septa. A- and B-ontogeny thus does not appear to be distinctly different in Buellia s. l. Among the specimens examined here, spores of B. prospersa are most easily recognized (Fig. 30). They are the largest spores and have a conspicuous septum thickening most distinctly observed in premature spores. This septum is persistent in mature spores, but eventually reduced in overmature spores. Because of the thickening, these spores have been referred to the Physconia-type (Scheidegger 1993). In some spores the septum remains conspicuously thickened during almost the entire ontogeny. Apical thickenings are never present and the spores could thus also be referred to as Orcularia-type. Mayrhofer et al. (1999) described a similarly persistent septum 246 thickening in Amandinea insperata (Nyl.) H. Mayrhofer & Ropin. Differences between the two types seem not very distinct, especially if apical wall thickenings cannot be observed in the Physconia-type. A faint and inconspicuous thickening can also be seen in the spore ontogeny of Buellia pullata, where it may easily be overlooked. These spores could thus be assigned alternatively to the Buellia (= Beltraminea)- as well as the Physconia-type (Fig. 30). A similar situation has been discussed for B. dispersa where the septum thickening is also restricted to a brief phase of the ontogeny (Bungartz et al. 2002). In spores of both B. prospersa and B. pullata a microrugulate ornamentation forms early. It is barely visible in premature spores, but becomes distinct in mature spores. Spores of B. christophii are very distinct in the premature stage when they are very broad to almost globose and have a conspicuously evenly thickened spore wall (Fig. 25E). The broad spore wall of these premature spores is also distinct in the TEM (Fig. 25F). Mature spores of the species become less globose and are more ellipsoid to oblong (Fig. 30). With spore maturity the conspicuously thickened wall disappears and spore ornamentation cannot be distinguished in any of the stages. Spores of B. sequax are often difficult to distinguish from B. pullata, B. tergua and B. ryanii. Premature and mature spores of B. sequax are distinctly narrowly oblong. Their shape is very characteristic but transient. Scheidegger (1993, p. 356) suggests that “... The type of B. abstracta combines a chasmolithic thallus and narrow spores and was thus previously thought to be a good species.” He continues to say that intermediate forms with B. sequax can be found and that B. sequax therefore constitutes a polymorphic 247 species. We have examined type material of both taxa (see the discussion on thallus variation). The spores are not very variable but basically follow the same ontogeny with a premature stage of characteristically narrow spores. This stage, however, is easily overlooked if the apothecium contains a majority of ellipsoid spores at a later stage of the ontogeny. Spore ornamentation of B. sequax develops early, but it is inconspicuous and can usually only be distinguished in overmature spores. Nordin (1997; 2000) emphasized spore ultrastructure of Buellia species with pluriseptate ascospores. In the species studied here, TEM yields little additional information. It is noteworthy, however, that in B. sequax spores with a fractured as well as a smooth perispore have been observed in the TEM (Fig. 26L,M). This emphasizes the dynamic development of the spore. Immature and premature spores have a smooth perispore (Figs. 26L and 30), which becomes fissured and eventually fractured with maturity (Figs. 26M and 30). Spores of B. ryanii and B. tergua usually become ± constricted along the septum (Fig. 30). This constriction is not obvious in young spores, but it is usually pronounced in overmature spores. Buellia ryanii has the smallest spores with no distinct ornamentation and thin spore walls. In contrast, spores of B. tergua have thicker walls, which early develop an ornamentation. The ornamentation is best seen in mature and overmature spores. Taxonomy.—Some of the species described here have bacilliform conidia (Figs. 25G, 26J, 27F,L and 31), others filiform conidia (Figs. 25M, 26E and 31). Otherwise all species are very similar. Indeed, this similarity is the main reason why the species were previously generally identified as B. punctata s. l. We can currently not confirm that any 248 saxicolous specimens from the Sonoran Desert Region belong to B. punctata s. str. According to Mayrhofer and Moberg (2002, p. 9), "saxicolous and muscicolous material usually placed under this name is much in need of taxonomic revision." Scheidegger (1993, p. 343) mentions that “Saxicolous material, usually placed under this name, is possibly not homogeneous and is not yet completely understood by the author ...” According to Scheidegger’s (1993) description, A. punctata s. str. has spores without a septum thickening, no spore ornamentation and short filiform conidia. Sheard and May (1997) noticed that the North American material, which they assigned to A. puncata, has ornamented spores, and a septum thickening can be observed at least during some short period of spore development. They also report that they have only seen a single specimen with short filiform conidia (averaging < 15 mm) and all other specimens have longer conidia. Sheard and May (1997) did not examine saxicolous material from the Southwest, and it is possible that the specimens identified as A. punctata do not strictly belong to that taxon (Sheard, pers. communication). From the taxa treated here, the newly described species are all distinct from B. punctata s. str. because of their short, bacilliform conidia (Figs. 25G, 27F,L and 31). In addition, B. ryanii and B. tergua are also characterized by having the apothecial pigment cinereorufa-green, that is absent from B. punctata s. str. The presence of broad, almost globose spores and a thick exciple also separates B. christophii from B. punctata s. str., even if conidia are not found in all of the specimens. The genus Buellia s. l. currently represents an amalgam of not necessarily closely related species and various attempts have been made to subdivide this large and 249 heterogeneous group into smaller more strictly defined genera. In this publication we have examined a group of very similar species, some of which could have been treated within the genus Amandinea, solely justified by the filiform conidia. Amandinea, however, does not currently constitute a more precisely defined taxonomic group than Buellia. When Scheidegger (1993) described Amandinea, he provided the first valid description for a genus previously suggested, but not validly published by Choisy (1950). Choisy (1950) distinguished Amandinea from Buellia because of the “pycnoconidies aciculaires”, i.e. the acicular conidia. Scheidegger (1993) emphasized the differences in conidial length and conidiophore structure as the only diagnostic characters of the new genus Amandinea. Choisy (1950) transferred Buellia coniops (Wahlenb.) Ach., B. myriocarpa (DC. ex I. M. Lamb. & DC.) De Not. and B. stigmatea Körb. into the new genus. Scheidegger (1993) selected A. coniops as the type of Amandinea and also included Rinodina lecideina and Buellia punctata. Subsequently, the genus became more widely accepted. Matzer et al. (1994) transferred Rinodina petermannii (Hue) Darb. into Amandinea, emphasizing its filiform conidia. The species investigated by Scheidegger (1993), as well as A. petermannii, are all saxicolous. Sheard and May (1997) transfered several corticolous Buellia and Rinodina species from North America into Amandinea. These North American species all have filiform conidia. Most recently Mayrhofer and Sheard(2002) transferred Rinodina cacuminum (Th. Fr.) Malme into Amandinea because of its filiform conidia, also citing “preliminary molecular results” (p. 440), that do not support the inclusion of that species in Rinodina. 250 Marbach (2000) argued that Sheard and May (1997) provided an emended genus concept for Amandinea, allowing the inclusion of species where no conidia could be observed. Bungartz and Nash (2004b) questioned this approach. They demonstrated that B. turgescens Nyl. ex Tuck., a species which Marbach (2000) included in Amandinea without having studied the pycnidia, is synonymous with B. badia (Fr.) A. Massal. Bungartz and Nash (2004b) showed, that Buellia badia is a species with bacilliform conidia, that therefore must not be included in Amandinea. There is currently little agreement about the characters characterizing the genus Amandinea. Scheidegger (pers. communication) suggests that Amandinea may also be characterized by the scarcity of lichen secondary metabolites in the thallus. Scheidegger (1993) did not detect any secondary metabolites in the species he included in the genus. Also, Sheard and May (1997) did not document any secondary metabolites in the North American species. Matzer et al. (1994), however, report norstictic acid and ergosterol peroxide from A. petermannii. Even though Scheidegger (1993) did not detect secondary metabolites in A. lecideina (= B. prospersa), that species is clearly characterized by a pale UV+ yellow to orange thallus, a reaction caused by various xanthones. Several species, which Marbach (2000) included in Amandinea, also have characteristic secondary metabolites. In addition, Marbach (2000) mentioned that Amandinea species often have a pale hypothecium, a character more commonly associated with Rinodina rather than Buellia. The species treated by Scheidegger (1993) (including the type) and most of the species treated by Sheard and May (1997), however, have a dark hypothecium. All species treated here, also have a dark brown hypothecium. 251 In his treatment of corticolous Buellia s. l., Marbach (2000) emphasized branching patterns of the paraphyses and the pigmentation of their apical cells as taxonomically important. According to Marbach (2000, p. 52) large, globose apical cells are characteristic for Amandinea. “Tafel 1/2: Paraphysenenden” (pp. 36 and 37, Marbach 2000), however, documents only very subtle, if any differences in the paraphyses of Buellia s. l. No distinct differences that would justify a clear segregation of the genus Amandinea, are displayed in this chart. Marbach (2000) did not discuss the different apothecial pigment types in any detail, even though these differences are clearly taxonomically important [e. g., in B. ryanii and B. tergua; see also: Bungartz and Nash (2004a)]. In addition to the paraphysal characters, Marbach (2000), p. 52) emphasized that spores without wall thickenings are typical for Amandinea. Nevertheless, he included several species within the genus, that have distinctly thickened septa. As previously discussed, wall thickenings vary considerably during spore ontogeny, and the spores of some species (like B. pullata) are difficult to assign to a particular type. Conspicuously thickened spore septa are characteristic for several species transferred from Rinodina into Amandinea. When Scheidegger (1993) validated Choisy’s genus description of Amandinea, he included two species with distinctly thickened septa, including the type A. coniops. By transferring R. insperata into Amandinea, Mayrhofer et al. (1999) also explicitly included a species with distinctly thickened spore septa. However, taxa transferred from Buellia (e. g., Buellia punctata) often lack distinct septum thickening. 252 In addition to the filiform conidia, Giralt et al. (2000, p. 521) provide a list of the following characters typical for Amandinea: “apothecia lecanorine or lecideine, ascospores brown, one-septate, Buellia- or Physconia-type, often with rugulate ornamentation; and thallus with norstictic acid or more frequently without secondary lichen compounds. ...” All these characters are regularly found in Buellia and by no means exclusive to Amandinea. A lecanorine apothecium is generally more common in Rinodina, but some species in Buellia also show the tendency to form a thalline exciple. In addition, immersed lecideine apothecia often emerge from the thallus with some thallus material remaining attached to the exciple (see B. ryanii and B. tergua). The ascus of Amandinea is generally described as the Bacidia-type (Marbach 2000; Giralt 2001), and it is thus not different from Buellia s. str. (Rambold et al. 1994). Søchting et al. (2004) recently transferred several arctic species from Buellia into Amandinea, arguing that in the absence of molecular data this transfer is only justified because of filiform conidia of those species. The filiform conidia borne on type III conidiophores in a Roccella-type pycnidium (sensu Vobis 1980) therefore represent the only reliable diagnostic character consistently used to seggregate Amandinea from Buellia. Within the Physciaceae, conidial length has also been used to distinguish Physcia from Phaeophyscia (Moberg 1977), Physcia from Hyperphyscia (Choisy 1950; Hafellner et al. 1979), and to characterize the genus Mobergia (Mayrhofer et al. 1992). None of these genera, however, are distinguished solely by the length of their conidia, and the foliose genera are also well circumscribed by a range of other characters. 253 The shape of conidia (bacilliform vs. filiform) is a result of conidial length and it does not represent a general difference in conidial structure. Conidial length may vary considerably even within the same pycnidium of a single specimen. In fact, B. pullata has predominantly 20-25 µm long, filiform conidia, but overall the conidia measured from a single pycnidium of that species vary from 6 to 43 µm in length (Fig. 31). The shortest conidia are thus bacilliform rather than filiform. In B. prospersa (= A. lecideina) the range of our measurements (7-32 µm, Fig. 31) considerably exceeds the variation observed by Scheidegger (1993), who measured conidia from 15-30 µm long. Scheidegger (1993, Fig. 5) plotted conidial length of the species in his treatment to demonstrate the differences between Buellia and Amandinea. The shortest conidia were found in B. vilis (2.5 – 4 µm). Buellia subdisciformis was the species with the longest conidia (9-14 µm) still treated within Buellia, but conidia of A. punctata were only slightly longer (up to 15 µm). Both A. lecideina and A. coniops had the longest conidia (15 – 30 µm). Plotting our measurements of B. pullata and B. prospersa (= A. lecideina) to this chart results in considerable overlap and conidial length can therefore no longer be regarded a discrete character. An asymptotic graph more accurately describes the situation (Fig. 31). In addition to the conidial length, differences in branching patterns of the hyphae bearing conidia can also be observed. Conidiophore structure is, however, closely related to the length of the conidia and cannot be treated as an independent character. Long, filiform conidia are generally borne on shorter and less extensively branched conidiophores. This is a necessary consequence of ejecting conidia from the pycnidium. 254 Long filiform conidia would become entangled in densely branched conidiophores. Densely branched conidiophores can, however, produce larger quantities of short, bacilliform conidia and can thus be found in species, which have only short conidia. In lichen taxonomy conidia alone have not generally been used to distinguish taxa at the generic level. In some groups, conidia of very different length can even be found on a single thallus. The genus Micarea regularly has both microconidia and macroconidia formed in different pycnidia on the same thallus. In other groups conidia of different length occur in closely related species of the same genus. For example, Lecanora himalayae Poelt (from Nepal) and two very similar species, L. chondroderma Zahlbr. (from China) and L. beamanii B. D. Ryan (from high elevations in southern Mexico) have straight, 6-8 µm long conidia. In contrast, L. maxima Lynge has curved conidia that are 11-17(-20) µm long. Nevertheless, all taxa belong to the section Dactylon Poelt that presumably forms a natural group isolated from other species of Lecanora (Ryan 1989). Other genera, for example Haematomma (Rogers & Hafellner 1988) or Aspicilia (Clauzade & Roux 1984), also include some species with bacilliform and others with filiform conidia. In the Physciaceae, the validity of using filiform vs. bacilliform conidia as the sole character to justify Amandinea has recently been questioned. Because of their filiform conidia, some terricolous species of the Buellia epigaea-group would have to be formally transferred into Amandina, even though they appear not closely related with other species with filiform conidia (Mayrhofer and U. Grube, pers. communication, Grube & Arup 2001; Trinkaus et al. 2001). Preliminary molecular studies also do not confirm 255 Amandinea as a monophylletic group (Mayrhofer pers. communication, Grube & Arup 2001; Wedin et al. 2002). In some lichen species pycnidia are very rare and are thus rarely found in all of the specimens examined. If pycnidia are rare, it may sometimes be difficult to assess if these conidia belong to a particular lichen specimen, or instead to a lichenicolous fungus growing inside this specimen. A character, that is frequently absent and difficult to assess does not seem to be particularly suitable as the sole diagnostic feature to circumscribe a genus. A monophylletic genus Amandinea with A. coniops as the type, will most likely not include a large portion of species currently included by various authors. In the Sonoran Desert, the B. mamillana-group (Bungartz, in prep.) includes species with xanthones that all have long bacilliform to fusiform conidia. Within Buellia s. l., that species group is isolated, but not closely related to species commonly treated within Amandinea. Because of its yellowish thallus containing xanthones, Buellia prospersa (= A. lecideina) shows some affinities with the B. mamillana-group. Buellia pullata, on the other hand, is fairly similar to the type species A. coniops, even though the spore septa are hardly thickened during their ontogeny. In summary, the taxonomy of Amandinea is not well resolved. Segregating the species treated here into separate genera only on the basis of conidial length would result in a constrained and arbitrary taxonomy. Nimis (1998) discussed three main criteria that justify the recognition of a genus in lichens: (1) monophylly, (2) phylogenetic analysis of its taxa, and (3) several independent characters that circumscribe the new genus. The 256 genus concept of Amandinea currently matches none of these criteria. “If only one character is involved, this is indeed the weakest possible evidence. It could easily be in conflict with the next character to be discovered...” (Nimis 1998, p. 432). This is exactly the situation in the genus Amandinea. Including species solely on the basis of conidial length does not help to resolve the taxonomy of Rinodina or Buellia, both of which remain large, artificial and heterogeneous genera. Several recent treatments (Grube & Arup 2001; Wedin et al. 2002; Helms et al. 2003) have confirmed two general clades in the Physciaceae: (1) the Physcia-(or Rinodina)clade, characterized by a Lecanora-type ascus, a usually hyaline hypothecium and spores regularly with wall thickenings (e. g., Physcia, Heterodermia, Anaptychia, Mobergia, Tornabea, Rinodina etc.) and (2) the Buellia-clade with Bacidia-(or Biatora)-type ascus, usually a dark apothecium and spores less frequently with wall thickenings (e. g., Buellia, Dimelaena, Dermatiscum, etc.). The “Caliciaceae” apparently also belong to this second clade (Wedin et al. 2002). The resolution within the clades is currently not sufficient to justify further segregation of the crustose genera Rinodina s. l. and Buellia s. l. Until a more resolved phylogeny is achieved, using both classical and molecular data, we advocate including the species currently treated as Amandinea within the genus Buellia s.l. Excluding species with filiform conidia from Buellia does not resolve the nomenclatural problems in this rather large and heterogeneous genus. According to the code of Botanical Nomenclature (Greuter et al. 2000), B. disciformis (Fr.) Mudd. is currently the listed type of the conserved genus Buellia. Unfortunately, the genus Hafellia 257 Kalb., H. Mayrhofer & Scheid. is also based on the same type. To avoid this conflict Moberg et al. (1999) suggested conserving Buellia with Buellia aethalea (Ach.) Th. Fr. as a new type. If this proposal is rejected, most species now treated within Buellia s.l. would then have to be accommodated in a new genus. This may include some of the species currently treated within Amandinea. The name Buellia s.str. would then refer to species generally treated as Hafellia, a genus which would become obsolete. None of the species in Amandinea is closely related to that group. If, however, the proposal by Moberg et al. (1999) is accepted, some of the species currently transferred into Amandinea, may eventually turn out to be more closely related to the B. aethalea-group and still better accommodated within Buellia. ACKNOWLEDGEMENTS We are obliged to Dr. Laurence A. Garvie, Department of Geology, Arizona State University (ASU), for access to a slow-speed diamond stone saw. Dr. Donald M. Burt, Dept. of Geology (ASU), identified the rock substrates of the specimens sectioned. The chemistry of all specimens was analyzed with TLC at ASU. Specimens from the private herbarium of Dr. Christoph Scheidegger were also examined with TLC by Martin Frei Swiss Federal Institute for Forest, Snow and Landscape Research (Birmensdorf). Additionally, selected specimens were analyzed with HPLC by Dr. Jack A. Elix from the Australian National University in Canberra. Dr. Ulrik Søchting, University of Copenhagen, Denmark, Julia Blaha and Ulrike Grube, University of Graz, Austria (GZU), contributed valuable information to the discussion of the taxonomy of Amandinea. Scott Bates (ASU) and Dr. John Sheard, University of Saskatchewan, 258 Canada, kindly reviewed first drafts of the manuscript. Dr. Helmut Mayrhofer (GZU) and an anonymous reviewer provided valuable input to the final version of this publication. Dr. Christian Printzen, Senckenberg Institut, Germany, was very helpful correcting the Latin diagnoses. Dr. Donald Pinkava, ASU, provided much support on taxonomic questions regarding the application of the ICBN. We would like to thank all herbaria, which have made specimens available for study, especially Julia Blaha (GZU), who – on short notice - supplied several specimens of B. prospersa, including the type of A. lecideina. On very short notice, Dr. Orvo Vitikainen, Helsinki (H) provided the Nylander specimen of B. prospersa used for lectotypefication. This study was supported by a Sigma Xi Grant-in-Aid of Research, sponsored by the local Sigma Xi chapter at ASU, and two National Science Foundation grants (DEB-0103738, DEB-9701111). LITERATURE CITED BELLEMÈRE, A. 1994. Asci and ascospores in ascomycete systematics. pp. 111-126. In HAWKSWORTH, D. L. (ed.) Ascomycete Systematics. Problems and Perspectives in the Nineties, NATO ASI Series, Series A: Life Sciences, 269. Plenum Press, New York. BUNGARTZ, F. & T. H. NASH III. 2004a. Buellia subalbula (Nyl.) Müll. 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British Lichen Society Bulletin 57: 1-41. 265 PUBLICATION 6 (The Bryologist, submitted February 2004): The Buellia aethalea-Group in the Greater Sonoran Desert Region with some reference to similar species known from North America F. BUNGARTZ AND T.H. NASH III Arizona State University Lichen Herbarium, School of Life Sciences, P. O. Box 87 4601, Tempe, AZ 85287–4601, U.S.A. e-mail: frank.bungartz@asu.edu; tom.nash@asu.edu ABSTRACT. Species related to Buellia aethalea have been examined from the Greater Sonoran Desert Region. In the region B. aethalea s.str. is a very rare montane species. A similar, subalpine species is B. eganii, first reported from New Mexico by R. S. Egan, now validly described here. Most common in the Buellia aethalea-group is B. spuria, a species which does, however, not occur in lowland desert areas of the Sonoran Region. Buellia stellulata is similar to B. spuria but differs in thallus chemistry and is restricted to areas along the coast. Although similar in thallus morphology, B. lacteoidea is not closely related to the Buellia aethalea-group. The new name B. maculata is introduced here for the illegitimate B. stigmaea. Buellia maculata is a species with a similar thallus but not closely related to the Buellia aethalea-group and absent from the North American Southwest. Buellia lepidastra shows some affinities to the group but like B. maculata is confined to eastern North America. 266 Species related to Buellia aethalea (Ach.) Th. Fr. are often regarded as the core group of the genus Buellia. They are potentially significant for the concept of the genus because of a proposal by Moberg et al. (1999) to change the listed type from B. disciformis to B. aethalea. This proposal has not yet formally been accepted or rejected. Species within the Buellia aethalea-group can only be included in Buellia s.str., if the proposal to list Buellia aethalea as the new type, is accepted. Otherwise a new genus name must be suggested to accommodate this group of species. Until a final decision is reached by the committee of fungi, a broad concept of the genus Buellia should reasonably be adopted. A suggestion to accommodate species from the Buellia aethaleagroup into a newly erected genus would inevitably cause confusion and is not reasonable unless the proposal by Moberg et al. (1999) is formally been rejected. Scheidegger (1993) revised the European species included in the Buellia aethaleagroup. Our revision of saxicolous species with one-septate ascospores from the Greater Sonoran Desert Region also includes several species informally included within that group. This paper focuses on these species and briefly discusses some species not found in the North American Southwest. In North America only three species can currently be included as part of the Buellia aethalea-group with some certainty: Buellia aethalea, B. spuria, and B. stellulata. Imshaug (1951) referred to these species as part of the “stirps spuria” and suggested that B. lacteoidea may also belong into the group. Buellia lacteoidea does, however, not appear to be closely related to Buellia aethalea because of several different characters, most notably spores with a distinctly thickened septum. Three other species will be 267 discussed here in the context of the Buellia aethalea-group: Buellia maculata, a species morphologically very similar to Buellia spuria, but without the characteristic aeruginose exciple pigment; Buellia lepidastra, a species with affinities to the Buellia aethalea-group not yet fully resolved; finally, B. eganii is described as superficially similar to B. aethalea but without an aeruginose exciple and lecanoric instead of norstictic acid. METHODS All specimens were examined with light microscopy using hand- and cryosections. Both conventional bright field microscopy (BF) as well as differential interference contrast (DIC) was used. Selected specimens were also studied with transmission electron microscopy (TEM) according to a protocol described in detail by Bungartz et al. (2002). To improve dehydration and infiltration this protocol has been modified according to Bungartz & Nash (2004a). All specimens were spot tested and routinely examined with standardized thin-layer chromatography (Culberson & Kristinsson 1970; Culberson & Johnson 1982; White & James 1985; Orange et al. 2001). TLC-plates were interpreted with the computer program WINTABOLITES Mietzsch et al. (1994), and scanned for permanent record Egan (2001). In addition a subset of specimens was analysed by Dr. Jack A. Elix from the Australian National University in Canberra using standardized high performance liquid chromatography (HPLC, Elix et al. 2003). Spore measurements are given according to Nordin (2000). Pigment names follow Meyer & Printzen (2000). 268 SPECIES DESCRIPTIONS BUELLIA AETHALEA (Ach.) Th. Fr., Lichenogr. Scand. 1: 604 (1874). Gyalecta aethalea Ach., Lichenogr. Univ.: 669 (1810). TYPE: UNITED KINGDOM. NORTHERN ENGLAND. Durham Co. 54°46'N, 1°34'W, ca. 67 m [original label data: 'Anglia, Durham'], Harriman s.n. (H–ACH 66! – lectotype selected by Foucard et al. 2002, UPS–ACH! – isolectotype, S – isotype). Buellia aethaleoides (Nyl.) H. Olivier Bull. Acad. Intern. Géogr. Bot 12: 176. 1903.— Lecidea aethaloides Nyl., Flora, Jena 68: 42. 1885. TYPE: FRANCE. PYRENÉES ORIENTALES. Amélie. 11 June 1884, Nylander s.n. (H-NYL 9280 – holotype). Rinodina atropallidula (Nyl.) Arnold, Flora, Jena 68: 236. 1885.—Lecanora atropallidula Nyl., Flora, Jena 55: 428. 1872. TYPE: FRANCE. PYRENÉES ORIENTALES. Força Real. 400 m, 16 July 1872, Nylander s.n. (H-NYL 28570 – holotype). Buellia nigerrima (Nyl.) Arn. Lich. exsicc. nr. 1780. 1899.—Lecidea nigerrima Nyl. in Sandst., Abh. naturw. Ver. Bremen 14: 491. 1898. TYPE: GERMANY. NIEDERSACHSEN. Oldenburg, on roof tiles of the two tile factoreis along the street from Zwischenahn to Edewecht, 53°10'N, 8°11'E, ca. 8 m [original label data: Oldenburg, auf Dachziegeln der beiden Ziegeleien an der Chaussee Zwischenahn-Edewecht] Sandstede s.n. (= Arn. Lich. exsicc. nr. 1780) (H-NYL 5795 – isotype, M! – holotype). Buellia ocellata var. tenella Müll. Arg., Flora, Jena 58: 62. 1875. TYPE: SWITZERLAND. VALAIS. Distelgrat, 1874, Brun s.n. (G – holotype). 269 Buellia baltica Erichsen, Verh. Bot. Ver. Prov. Brandenburg 72: 46. 1930. TYPE: GERMANY. SCHLESWIG-HOLSTEIN. Kreis Plön. Hohwacht, on immersed stones of scree dunes near Strandesberg, supralittoral zone, 54°19'N, 10°40'60"E, ca. 0 m [original label data: an eingebetteten Steinen der Gerölldünen bei Strandesberg, supralittoral zone], 29 August 1933, Erichsen s.n. (HBG – lectotype selected by Scheidegger 1993, S! – isolectotype); Kreis Ekernförde, an Blöcken am Strande bei Aschau, 21 September 1924, Erichsen s.n. (= Lich. Exs. P. Vrang 113, S! – syntype). Note: The specimen mentioned by Scheidegger (1993) as holotype is effectively a lectotype because the protologue by Erichsen indiscriminantly mentions several specimens. Buellia sororia Th. Fr., Lichenogr. Scand. 1: 603. 1874. TYPE: SWEDEN. SÖDERMANLAND. Vöstermo prästgård, 1872, Blomberg s.n. (UPS – lectotype selected by Scheidegger 1993). Rinodina ocellulata Bagl. & Carest., Atti Soc. Crittog. Ital. 2: 210. 1880. TYPE: ITALY. LOMBARDIA. Vallesia (= Wallis). Varallo. On an arid wall made of diorit, 45°49'N, 8°15'E , ca. 440 m [original label data: Varallo (Valesia), su di un muro a secco fatto con pietre dioritiche alle scarpie di Valmaggia], 1877, Carestia s.n. (= hb. Critt. Ital. Ser 2: no. 721) (MOD – isotype, M-0023752! – isotype). Buellia sororioides Erichsen, Verh. Bot. Ver. Prov. Brandenburg 72: 49. 1930. TYPE: GERMANY. SCHLESWIG-HOLSTEIN. Kreis Lauenburg. On erratic boulders near Buchhorst, 53°22'N, 10°34'E, ca. 8 m [original label data: Kreis Lauenburg. An erratischen Blöcken bei Buchhorst], 3 October 1926, Erichsen s.n. (HBG – lectotype); Kreis Eckernförde. An fränkischen Steingräbern am Strande bei Hemmelmark, 20 July 270 1925, Erichsen 18 (= Lich. Exs. P. Vrang 23, S! – syntype; US! – syntype); Kreis Plön. Hohwacht bei Strandersberg. An Geröll am Strand, 1933, Erichsen s.n. (S! – syntype). Note: The specimen mentioned by Scheidegger (1993) as holotype is effectively a lectotype because the protologue by Erichsen indiscriminantly mentions several specimens. Buellia sororioides f. dendritica Erichsen, Verh. Bot. Ver. Prov. Brandenburg 72: 49. 1930. TYPE: GERMANY. SCHLESWIG-HOLSTEIN. Kreis Flensburg. Angeln. On scree along the beach near Birknach [original label data: auf Geröll am Strande bei Birknach], 21 September 1914, Erichsen s.n. (HBG – holotype). Buellia subatra Erichsen, Hedwigia 70: 218. 1930. TYPE: GERMANY. SCHLESWIGHOLSTEIN. Kreis Lauenburg. Growing abundantly on a wall of boulders west of Kasseburg, along the way to Friedrichsruh, 53°34'N, 10°24'E ca. 43 m [original label data: an einem Blockwall westl. von Kasseburg, am Wege nach Friedrichsruh, in Menge], 1 April 1927, Erichsen s.n. (HBG – holotype; S! – two isotypes). For additional synonyms see Foucard et al. (2002). Thallus (Fig. 32A,B) crustose, thin, ± continuous, epilithic; areolate; hypothallus usually distinct, black, between areoles and surrounding the thallus; thallus surface matt and dull, not shiny, usually gray to pale brown, rarely dark gray, epruinose, phenocorticate; lacking Ca-oxalate crystals within thallus medulla (H2SO4–, not forming clusters of needle-shaped crystals). Apothecia lecideine; (0.1–)0.2–0.3(–0.5) mm in diameter; remaining immersed, not becoming sessile, predominantly in the center of an 271 areole; proper margin indistinct, reduced, inconspicious, black; disc black, epruinose, plane, not becoming convex with age; exciple narrow, poorly differentiated, of the aethalea-type (Fig. 32C) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), often reduced, similar in structure and orientation to the paraphyses, transient with the hyaline to pale brown hypothecium (textura intricata); outer excipular hyphae parallel, moderately swollen (textura oblita) and usually strongly carbonized with various amounts of brown and aeruginose pigments (cf. elachista-brown & cinereorufa-green, HNO3+ violet), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with brown pigment cap (cf. elachista-brown) and diffuse, aeruginose pigment (HNO3+ violet, cinereorufa-green). Asci 8-spored, clavate, Bacidiatype. Ascospores (Fig. 32D,E) broadly ellipsoid, constricted with age, with obtuse ends, not curved, (11.0–)11.6–[12.8]–14.1(–17.0) x (5.0–)7.2–[8.1]–8.9(–10.0) µm (n = 60); one-septate, proper septum narrow not thickened during spore ontogeny, lateral wall thickenings absent [Beltraminea(=Buellia)-type]; ornamentation microrugulate. Pycnidia rare, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicariatype (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 5.0–5.5 x < 1.0 µm (n = 20). Chemistry.—All Sonoran specimens are characterized by the presence of only two depsidones: norstictic and connorstictic acid. Other substances are absent. The thallus and 272 medulla react K+ yellow to red (forming orange crystals if observed in the compound microscope), P+ yellow, C–, KC–, CK–. UV– (dark). The thallus is not amyloid [at least → FIGURE 32. Buellia aethalea (light micrographs).—A. Areolate thallus with immersed apothecia (Ryan 23545).—B. Close-up of areoles with irregular, predominantly central apothecia (Ryan 23545).—C. Cross section of apothecium (arrows indicate the reduced aethalea-type exciple; Nash 25427).—D. Hymenium with asci, paraphyses and ascospores (Ryan 23545).—E. Microrugulate mature ascospores (Ryan 23545). 273 274 in all Sonoran specimens, but this reaction is variable according to Scheidegger (1993)]; apothecia react amyloid in Lugol’s (always test the thallus reaction with concentrated iodine or in the compound microscope; positive reactions can be very weak!). Substrate and ecology.—On a variety of hard, siliceous (HCl–) rock substrates. Distribution (Fig. 7).—Rare, in the Sonoran region currently known only from a few localities at higher elevation. Notes.—All Sonoran specimens have a hyaline to faintly brown hypothecium. Scheidegger (1993) reports that hypothecium pigmentation of European specimens varies from hyaline to dark brown. This is rather unusual because Buellia s.str. is generally characterized by a dark hypothecium. Scheidegger (1993) also reports that the medullary reaction with Lugol’s iodine varies considerably. All Sonoran specimens are, however, I–. Additional specimens examined.–ARGENTINA. CATAMARCA. Lamb 5587 (MSC340260). FRANCE. PYRENÉES ORIENTALES. Scheidegger Inv. Nr. 7990, 8293, 8092 (hb. Scheidegger). GERMANY. NIEDERSACHSEN. Kreis Gosslar Co. Imshaug 36300 (MSC76545); Imshaug 36300 (MSC-76545), Ullrich s.n. (MSC-67311). SWEDEN. VÄRMLANDS LAN. Sundell sn. (MSC-40864). SWITZERLAND. WALLIS. Scheidegger Inv. Nr. 8554 (hb. Scheidegger). SWEDEN. SKÅNE. Almborn s.n. (ASU). U.S.A. ARIZONA. Apache Co. Nash 26943 (ASU). Cochise Co. Weber S28044a (ASU). Coconino Co. Nash 25427b (ASU). PIMA CO. Nash 20716 (ASU). Santa Cruz Co. Nash 18583 (ASU); Scheidegger field notes 31-39 (hb. Scheidegger). COLORADO. Clear Creek Co. Anderson 5292 (ASU). 275 MICHIGAN. Unknown Co. Harris 7687 (MSC-127522). SOUTH DAKOTA. Custer Co. Wetmore 10338 B (MSC-68823). Lawrence Co. Wetmore 8506 (MSC-69496). Pennington Co. Wetmore 3451 (MSC-68965); Wetmore 7221 (MSC-68960); Wetmore 7907 (MSC68958). U.S.A.. WYOMING. Crook Co. Climbers 11423 A (MSC-69773). BUELLIA EGANII Bungartz sp. nov. Thallus saxicolus, crustaceus, areolatus, crassus, griseus vel badius, fulvus. Hypothallus atratus. Apothecia immersa, lecideina, marginibus propriis tenuibus. Excipulum tenue, fulvo-caeruleum, pigmentum aeruginosum deficiens, carbonaceum. Asci 8-spori. Sporae uniseptae, ellipsoideae vel oblongae, 12–18 x 6–9 µm. Thallus acida lecanorica continens. Similis Buelliae athaleae sed apothecibus lateralis, thallus crassus et materia chimica differt. TYPE: U.S.A. NEW MEXICO. Santa Fe Co. Lake Peak, 18 km NE of Santa Fe, 35°47'47"N, 105°46'14"W; alpine tundra; 3750 m elevation; 14 August 1969; Egan El1672 (OMA! – holotype designated here). The species is named in honor of Dr. Robert S. Egan, University of Nebraska at Omaha. He first discovered B. eganii working on his PhD dissertation. In his dissertation Egan called the species B. lakensis referring to the type locality Lake Peak, New Mexico. His description was never validly published. 276 Thallus (Fig. 33A,B) crustose, moderately thick, ± dispersed, epilithic; areolate to subsquamulose; hypothallus absent or, if present, indistinct and not prominent; thallus surface ± shiny and smooth, or matt, usually pale beige, rarely pale gray, epruinose, → FIGURE 33. Buellia eganii (light micrographs).—A. Areolate thallus with immersed apothecia (Nash 25427).—B. Close-up of areoles with irregular, predominantly marginal apothecia (Nordin 5311).—C. Cross section of apothecium (arrows indicate the reduced aethalea-type exciple; Nordin 5311).—D. Microrugulate mature ascospore (Nordin 5311).—E. Photobiont cells of Buellia eganii (arrows indicate single cells of the alga Trebouxia. sp.; Nordin 5311). 277 278 phenocorticate; without calcium oxalate crystals in the medulla (H2SO4–). Apothecia lecideine; (0.2–)0.3–0.5(–0.7) mm in diameter; remaining immersed, not becoming sessile, predominantly along the margin of an areole; proper margin indistinct, reduced, inconspicious, black; disc black, epruinose, plane, rarely becoming slightly convex with age; exciple narrow, poorly differentiated, of the aethalea-type (Fig. 33C) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), often reduced, similar in structure and orientation to the paraphyses, transient with the hyaline to faintly brown hypothecium (pigmentation ± absent, textura intricata), outer excipular hyphae parallel, moderately swollen (textura oblita) and usually strongly carbonized with various amounts of brown pigments (cf. elachistabrown, HNO3–), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown). Asci 8-spored, clavate, Bacidia-type. Ascospores (Fig. 33D) oblong to ellipsoid, usually not constricted, with obtuse ends, not curved, (12.0– )12.1–[13.7]–15.2(–18.0) x (6.0–)6.3–[7.0]–7.7(–9.0) µm (n = 60); one-septate, proper septum narrow, not thickened during spore ontogeny [Beltraminea(=Buellia)-type]; ornamentation microrugulate. Pycnidia rare, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 4.0–7.0 x < 1.0 µm (n = 20). 279 Chemistry.—With the monocyclic phenol derivate lecanoric acid and the depside 5-O-methylhiascic acid (confirmed by HPLC, J. Elix, Canberra). Thallus and medulla C+ pink (fleeting), KC+ pink (fleeting) or C–, KC–, K–, P–. UV+ pale beige to yellow. The thallus (cortex and medulla) is not amyloid; apothecia react amyloid in Lugol’s (always test the thallus reaction with concentrated Lugol's iodine or in the compound microscope; positive reactions can be very weak!). Substrate and ecology.—On a variety of siliceous (HCl–) rock substrates in subalpine to alpine habitats. Distribution (Fig. 8).—Currently only known from Lake Peak and Sierra Blanca Peak (New Mexico), Mt. Humphries (San Francisco Peaks, Arizona) and the White Mountains (Arizona) Notes.—The species is morphologically and anatomically very similar to B. aethalea but because of the different chemistry and the absence of cinereorufa-green possibly not closely related. Additional specimens examined.–U.S.A. ARIZONA. Apache Co. Nash 11720 (ASU). Coconino Co. Nash 36934. New Mexico. Santa Fe Co. Egan El-2532 (OMA). Otero Co. El-1747 (OMA). BUELLIA LACTEOIDEA de Lesd., Ann. Crypt. Exot. 5: 129. 1932. TYPE: U.S.A. NEW MEXICO. San Miguel Co. Hermit’s Peak, 35°44'39" N, 105°24'52"W; on siliceous rock [original label data: Hermit’s Peak, roche siliceuse], 3110 m elevation, 2 July 1930, Frère Arsène Brouard s.n. (W! – lectotype selected here; 280 S!, UPS! – isolectotypes). Note: The lectotype has been selected here because it can be assumed that the holotype material was destroyed in World War II. Thallus (Fig. 34A,B) crustose, thin to moderately thick, ± dispersed, epilithic; areolate; hypothallus conspiciously black, frequently strongly developed and usually between the areoles as well as surrounding the thallus outline; thallus surface matt and dull, not shiny, white to whitish gray, rarely beige, epruinose; phenocorticate; without Ca-oxalate crystals in the medulla (H2SO4–). Apothecia lecideine; (0.2–)0.3–0.6(–1.1) mm in diameter; remaining immersed, not becoming sessile (but disc sometimes convex and emerging from the surface); proper margin indistinct, reduced, inconspicious, black or rarely masked by a thalline veil; disc black, epruinose, plane, often becoming strongly convex with age; exciple narrow, poorly differentiated, of the aethalea–type (Fig. 34C,F) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), often reduced, similar in structure and orientation to the paraphyses, transient with the hyaline subhymenium and the brown hypothecium (leptoclinoides–brown, textura intricata), outer excipular hyphae parallel, moderately swollen (textura oblita) and usually strongly carbonized with various amounts of brown and aeruginose pigments (cf. elachista-brown & cinereorufa-green), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown) and a diffuse, aeruginose pigment (HNO3+ violet, cinereorufagreen). Asci 8-spored, clavate, Bacidia-type. Ascospores (Fig. 34D,E,G,H) oblong to 281 ellipsoid, usually not constricted, with obtuse ends, not curved, (12.0–)13.5–[15.1]– 16.7(–20.0) x (6.0–)6.2–[7.0]–7.9(–9.0) µm (n = 60); one-septate, proper septum soon but only briefly thickened during spore ontogeny (Physconia-type); ornamentation rugulate; septum with septal pore canal, simple pore and undifferentiated pore plug; spore wall (Fig. 34G, H) differentiated into cracked, thin perispore (0.10–0.15 µm), narrow intermediate layer (< 0.06 µm), thick proper spore wall (0.30–0.47 µm) and thick endospore (0.30–0.57 µm). Pycnidia rare, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 2.5–4.5 x 1.5–2.0 µm (n = 20). Chemistry.—Generally with the depside atranorin and the depsidones norstictic and connorstictic acid. Additionally often with gyrophoric acid (monocyclic phenol derivate). The following HPLC–artefacts were also found: haematommic acid, methyl β-orsellinate (HPLC by J. Elix, Canberra). The thallus and medulla usually react K+ yellow to red (forming needle-shaped orange crystals if observed in the compound microscope), P+ yellow, C– (rarely C+ pink), KC– (rarely KC+ pink), CK–. UV+ pale ivory (if gyrophoric acid is present). The medulla reacts very strongly amyloid and the reaction is even visible if specimens are tested directly on the thallus cortex; apothecia react amyloid in Lugol’s. Substrate and ecology.—On a variety of hard siliceous (HCl–) rock substrates. 282 → FIGURE 34. Buellia lacteoidea [A–F. light micrographs (Ryan 10885), G–H. TEM micrographs (Nordin 5162)].—A. Areolate thallus with immersed apothecia.—B. Closeup or areoles with irregular, predominantly marginal apothecia.—C. Cross section of young apothecium with hyaline subhymenium (sh).—D. Ascus with immature ascospores.—E. Premature ascospore with a ± thickened septum.—F. Cross section of older apothecium with a hyaline subhymenium (sh) and dark hypothecium (hyp).—G. Mature ascospore.—H. Spore wall of mature ascospore: (as) ascus wall; (1) perispore; (2) intermediate layer; (3) proper spore wall; (4) endospore. 283 284 Distribution (Fig. 8).—Only known from western North America; not a typical desert species but frequent at higher elevation (montane). Notes.—Buellia lacteoidea has often been confused with B. spuria. The two species are superficially very similar. Imshaug (1951) suggested that the species can be distinguished by the hyaline hypothecium of B. lacteoidea. This suggestion is misleading. Buellia lacteoidea is the only species currently known from the Sonoran Region with a differentiation of a hyaline subhymenium and a pigmented hypothecium. Although the hypothecium of young specimens of B. lacteoidea is only faintly colored, this pigmentation becomes stronger with age. If mature apothecia are sectioned for comparison, B. lacteoidea may thus easily be confused with B. spuria. The species can, nevertheless, easily be distinguished if not only the hypothecium but also the spore ontogeny and the position of the apothecia are compared. In B. lacteoidea apothecia generally remain immersed and often become deformed by adjoining areoles. Young apothecia of B. spuria are also immersed but eventually emerge from the thallus and become distinctly sessile. B. lacteoidea has spores with a brief phase when a distinctly thickened median septum can be distinguished. No wall or septum thickenings are present during the spore ontogeny of B. spuria. Both species have a medulla reacting with Lugol’s iodine, but the reaction is much stronger and more persistent in B. lacteoidea. Additional specimens examined.–MEXICO. BAJA CALIFORNIA. Nash 14739 (ASU). U.S.A. ARIZONA. Apache Co. Hertel 40155, 40181, 40175, 40138 (M); Nash 27025a, 34846, 11783, 11794, 10034a, 10034b, 10754b (ASU). Cochise Co. Darrow 4208, Nash 14579, 14586, 15637, 3888 , 9653a , Ryan 10877, 10885, 11002 (ASU); Hertel 40070, 285 40045 (M). Coconino Co. Nash 25426, 36933, 8403 (ASU); Hertel 40297, 40301, 40244 (M). Gila Co. Nash 6649 (M). Graham Co. Nash 16630 (ASU). Greenlee Co. Nash 10642b (ASU). Pima Co. Nash 27435, 20714, 4075, 40016 (M); Hertel 40013 (M); Wetmore 54553 (MIN). Santa Cruz Co. Darrow 4368, Nash 18582, 8282, 29993 (ASU). COLORADO. Boulder Co. Anderson 9287 (ASU). NEW MEXICO. Catron Co. Nash 22588 (ASU). Sierra Co. Nash 7110 (ASU). San Miguel Co. Imshaug 10016 (MSC); Imshaug 9940 (MSC-335171); Imshaug 9928 (MSC-335156); Imshaug 9930 (MSC-335157). SOUTH DAKOTA. Custer Co. Wetmore 7219 (MSC-65895); Wetmore 8281 (MSC-65940). Pennington Co. Wetmore 6450 (M-0061340); Imshaug 8034 A (MSC-65968); Wetmore 8028 A (MSC-65970); Wetmore 6450 (MSC-65837). UTAH. Garfield Co. Nebecker 2166 (ASU). BUELLIA SPURIA (Schaer.) Anzi, Cat. Lich. Sondr.: 87. 1860. Lecidea spuria Schaer., Lich. Helvet. Spicil. Sect. 3: 127. 1828. TYPE: SWITZERLAND. KANTON ZÜRICH. On granitic alpine boulders growing with Lecidea atro-alba [original label data: Ad saxa granitica, Schleicher, 1823 sub Lecidea atro-alba; Hepp, Flechten Europas Nr. 33, auf Alpenfindlingen bei Zürich], collected by Schleicher s.n., distributed by Hepp, Lich. Helvet. Exs. 33 (BERN – neotype, selected by Scheidegger 1993; M-0023903! – isoneotype, BM-000660170! – isoneotype). Note: Specimen BM-000660170 is mounted on the same sheet with BM-000660171, which, however, has different spores and is not part of the type. 286 Buellia lactea (A. Massal.) Körb., Parerg. Lich.: 183. 1860.—Buellia italica var. lactea A. Massal, Schedul. Critic. 9:193. 1856.—Catolechia lactea A. Massal., Ricerch. Auton. Lich.: 84. 1852. TYPE: ITALY. VENETO. Monte Bolca. On basalt, 45°36'N, 11°11'E, ca. 791 m [original label data: ad basaltica], 1849, Massalongo s.n. (MOD – holotype). Buellia olivaceofusca (Anzi) Zahlbr., Cat. Lich. Univ. 7: 385. 1931.—Buellia lactea var. olivaceofusca Anzi, Atti. Soc. Ital. Sci. Natur. 9: 252. 1866. TYPE: ITALY. TOSCANA. On mica-schist from the mountain Pisano [original label data: Sul micaschisto nel monte Pisano] Anzi s.n. (TO – holotype, W – isotype). Buellia italica var. recobarina A. Massal., Sched. Crit. 9: 163. 1865. TYPE: ITALY. Living on rock in the vicinity of Voltri [original label data: Vive sulli rupe nelle vicinanze di Voltri], Massalongo s.n. (W – isotype). Buellia italica var. insularis Bagl., Nuov. Giorn. Bot. Ital. 3: 264. 1871. TYPE: ITALY. SARDINIA. Orri, Baglietto s.n. (G – isotype). Buellia exilis (Kremp.) Müll. Arg., Flora 70: 61. 1887.—Lecidea exilis Kremp., Verh. zool.-bot. Ges. Wien 26: 444. 1876. non Kremp. Verh. zool.-bot. Ges. Wien 30: 340. 1880. TYPE: PERU. LIMA. Circumnavigation of the World by S.M. Donau 1868-1871, Barrania s.n. (M-0023771!). Note: The type material is a specimen named by Krempelhuber as Lecidea excilis Kremp. (Verh. zool.-bot. Ges. Wien 30: 340. 1880). Mayrhofer recognized the name as illegitimate and annotated the specimen as Buellia krempelhuberi Zahlbr. (vidi H. Mayrhofer, Graz, 1983). The specimen was examined by 287 Elix & Wrardlaw with HPLC and contains: atranorin (major), chloroatranorin (major), norstictic acid (minor), stictic acid (major), cryptostictic acid (minor). Buellia liguriensis de Lesd., Bull. Soc. Bot. France 101(5-6): 225. 1954. TYPE: ITALY. LIGURIA. Spotorno. On rock. [original label data: Liguria occid.: Spotorno; rupicola, specim. orig.]; January 1953, Sbarbaro s.n. (US! – lectotype selected here). Note: The holotype was probably destroyed in World War II. Buellia norstictica Imshaug ined. Note: Name never validly published. Two specimens deposited in M collected by Imshaug & Harris on the East Falkland Islands belong to Buellia spuria (UTM Grid 21F VC 4373, Port William: Outcrops on Engineer Point headland, The Narrows, 50 ft. 19 January 1968. Imshaug 40617; UTM Grid 21F VC 3472, Stanley: Empetrum-heath & outcrops on Tumbledown Mountain, 500-700 ft. 3 January 1968, Imshaug 39675). Thallus (Fig. 35A,B) crustose, thin to moderately thickened, ± continuous, epilithic; areolate; hypothallus conspiciously black, in most specimens strongly developed and growing between the areoles, rarely only surrounding the thallus outline; thallus surface matt and dull to ± shiny, usually white to whitish gray, rarely dark gray, epruinose, phenocorticate; without crystals in the medulla (H2SO4–). Apothecia lecideine; (0.2–)0.3– 0.4(–0.5) mm in diameter; immersed to adnate, rarely sessile; proper margin thin, ± persistent, rarely excluded with age, black or color masked by grayish remains of necrotic thalline material (thalline veil); disc black, epruinose, plane, rarely becoming slightly convex with age; exciple narrow, poorly differentiated, of the aethalea-type (Fig. 35C) 288 sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), often reduced, similar in structure and → FIGURE 35. Buellia spuria [A–D. light micrographs (Nash 10755), E–I. TEM micrographs (Nash 35879)].—A. Areolate thallus with immersed to slightly sessile, circular apothecia.—B. Close-up of areoles with ± immersed apothecia, some with a thalline veil (arrows).—C. Cross section of an apothecium.—D. Mature ascospores.—E. Immature ascus.—F. Immature ascospore: the endospore and proper spore wall not yet fully differentiated (1), but the perispore (2) is clearly visible; (as) ascus wall.—G. Mature ascospore.—H. Bacidia-type ascus (for designation of the different layers see Bellemère 1994): The a- and b-layer (ab) are barely visible and cannot reliably be distinguished (possibly as a result of fixation artefacts); (c) outer electron opaque c-layer; (d1) d1-layer, i.e. the outer tholus, which is distinctly laminated (in light microscopy this outer part stains deep blue with Lugol’s iodine); (d2) d2-layer, i.e. the inner tholus, which is not layered and ± homogeneous (not staining in Lugol’s iodine); (oc) ocular chamber.—I. Spore wall of a mature ascospore: (as) ascus wall; (1) perispore; (2) intermediate layer; (3) proper spore wall; (4) endospore. 289 290 orientation to the paraphyses, transient with the deep reddish brown hypothecium (leptoclinoides-brown, textura intricata), outer excipular hyphae parallel, moderately swollen (textura oblita) and usually strongly carbonized with various amounts of brown and aeruginose pigments (cf. elachista-brown & cinereorufa-green), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachistabrown) and a diffuse, aeruginose pigment (HNO3+ violet, cinereorufa-green). Asci 8spored, clavate, Bacidia-type (Fig. 35E,H). Ascospores (Fig. 35D,F,G) oblong to ellipsoid, usually not constricted, with obtuse ends, not curved, (11.0–)11.9–[13.3]– 14.8(–18.0) x (5.0–)5.9–[6.7]–7.4(–8.0) µm (n = 60); one-septate, proper septum narrow not thickening during spore ontogeny [Beltraminea(=Buellia)-type]; ornamentation microrugulate, or absent in some specimens; septum with septal pore canal, simple pore and undifferentiated pore plug; spore wall (Fig. 35I) differentiated into smooth to fissured, thin perispore (0.18–0.23 µm), narrow intermediate layer (< 0.03 µm), thick proper spore wall (0.32–0.50 µm) and thin endospore (0.10–0.16 µm). Pycnidia rare, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicariatype (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 4.5–6.0 x 0.5–1.0 µm (n = 20). Chemistry.—The biosynthetically related depsides atranorin and chloroatranorin and artifacts generated by the hydrolysis of atranorin (haematommic acid and methyl β- 291 orsellinate) have been detected by HPLC. In addition the depsidones norstictic with connorstictic acid are regularly present and several other depsidones can also be reported: stictic with constictic acid and cryptostictic, hypostictic and methylstictic acid have been detected (HPLC by J. Elix, Canberra). The thallus usually reacts K+ yellow to red (orange crystals forming in the compound microscope), P– or + yellow, C–, KC–, CK–. UV– (dark). The thallus is amyloid (test the medulla, not the cortex) and the apothecia also react amyloid in Lugol’s. Substrate and ecology.—On a variety of siliceous (HCl–) rock substrates. Distribution (Fig. 10).—Common and widely distributed throughout the Northern Hemisphere. In the Sonoran Region not a typical desert species but frequent at higher elevation (montane). Notes.—For comparison with B. lacteoidea see the notes on that species. Buellia stellulata is morphologically very similar but consistently has a non-amyloid medulla and contains 2-O'-methylperlatolic acid instead of norstictic acid. Additional specimens examined.—ARGENTINA. CATAMARCA. Lamb 5525 (MSC354199); Lamb 5504 (MSC-340263); Lamb 5525 (MSC-356199); Lamb 5646 (MSC340261). TUCUMAN. Lamb 5354 (MSC-). CUBA. LA HABANA. Imshaug 24742 (MSC348151); Imshaug 25013 (MSC-348113). FRANCE. LANGUEDOC. Vezda s.n. (MSC77231). ITALY. LIGURIA. Sbarbaro s.n. (MSC-146477); Sbarbaro s.n. (MSC-146480). MEXICO. BAJA CALIFORNIA. Nash 34365, 38367, 8688, 26041 (ASU). BAJA CALIFORNIA SUR. Nash 30404, 12656, 12663, 12762, 39767, 39836, 29731b (ASU). CHIHUAHUA. Nash 31199, 36168, 36255, 36477, 36636, 37522, 37724, 13748, 13813, 13721b (ASU). 292 DURANGO. Nash 31177, 13929, 13936, 14049b (ASU). SINALOA. Nash 10145, 12151, 12169, 12220, 12087b (ASU). SONORA. Wetmore 71688 (MIN); Nash 30239, 30353, 33600, 37952, 10876, 10932, 10969, 10975, 11951a, 12013, 12441, 10959e; Ryan 21626, 21642 (ASU). U.S.A. ARIZONA. Apache Co. Hertel 40116 (M); Nash 10545a, 10554, 10588a, 10602, 10608, 10612, 27025b (ASU). Cochise Co. Hertel 40054, 40020 (M); Nash 3663a, 3723, 3824, 6968, 9679, 9687, 3663c, Ryan 10727, 10956, 10987, Zambrano s.n. (ASU). Coconino Co. Zschau s.n. (ASU). Gila Co. Biringer 115 (ASU). Graham Co. Nash 36053, 36084, 14479, 4197 (ASU). Greenlee Co. Nash 10642a (ASU). Navajo Co. Nash 10290a, 11185 (ASU). Pima Co. Biringer 40, Gebauer s.n., Kantrud 179, Nash 4120, 6422, Ryan 22321, 22322b (ASU); Hertel 39914a (M). Santa Cruz Co. Biringer 79, Darrow 489, 4255, 4259, 811, Nash 22277, 13024, 18583, 18588, 18647, 6080, 8568, 8584, 13008b (ASU); Hertel 39756 (M). Yavapai Co. Nash 6337 (ASU). COLORADO. Jefferson Co. Weber 18939 (MSC-54263); Erdman 18939 (MSC-54263). LOUISIANA. Natchitoches Co. Tucker 17480 (SBBG, MSC-141381). MONTANA. Gallatin Co. Nash 21672 (ASU). NEW MEXICO. Socorro Co. Nash 22538 (ASU). TEXAS. Brewster Co. Wetmore 19408, 18467 (MIN). SOUTH DAKOTA. Custer Co. Wetmore 7583 B (MSC69458). TEXAS. Jeff Davis Co. Nash 6563, 6584 (MIN). WASHINGTON. Whatcom Co. Ryan 7856 (ASU). 293 BUELLIA STELLULATA (Taylor) Mudd, Manual Brit. Lich.: 216. 1861. Lecidea stellulata Taylor, in Mack. Flora Hibernica 2: 118. 1836. TYPE: IRELAND. Kerry Co. Kerry Mountains, possibly a mountain near Carrig East, 51°55'N, 9°37''W [original label data: Carig Mountain] Taylor ex hb. Borrer s.n. (BM000660158! – lectotype selected by Foucard et al. 2002). Note: Scheidegger (1993) referred to a specimen from BM as the holotype. Foucard et al. (2002, p. 71) correctly suggested that “Taylor cites more than one specimen and a lectotype must be selected.” Unfortunately their lectotype designation is inaccurate. The spelling “Craig Mountain” was never used by Taylor and no such place apparently exists in Ireland (S. Louwhoff, lichen curator at BM, pers. comm.). Taylor (1836) wrote in the protologue of the species: “...on silicious and aluminious rocks in the Kerry Mountains, also near the sea shore [...]. Probably this species is alluded to by Mr Borrer under Verrucaria polysticta in Eng. Bot. Sup. t. 2741.” A sheet from BM with this specimen collected by Borrer (BM-000660158) is annotated in Taylor’s handwriting as “Lecidea stellulata Tayl. Carig Mountain from Dr. Taylor in herb. Borrer”. It can reasonably be assumed that Taylor regarded this specimen as the type. Research for the type locality yielded little conclusive information. The original locality information possibly refers to a mountain near the small village East Carrig, northeast of the city of Kenmare in the Kerry Mountains, County of Kerry. Buellia candidula Arnold, Verh. zool. bot. Ges. Wien 22: 291. 1872. TYPE: ITALY. TRENTINO (= SOUTH TIROL). On boulders above Gries [original label data: An Blöcken oberhalb Gries], Arnold s.n. (M – holotype). 294 Buellia microstellulata Imsh. ined. Note: Imshaug never published this name. The specimens at MSC from South America (Juan Fernandez) belong to B. stellulata. Buellia rinodina Malme ined. Note: A specimen in Krempelhuber’s herbarium no. 3847 (M-0061371!) is annotated as “Buellia rinodina Malme (Lecidea squamulata Fee)” but this name was apparently never published. The specimen was collected by Glaziou in 1869 in Brasil, Rio de Janeiro. It clearly belongs to B. stellulata. The following species have been treated as synonyms of B. stellulata but do not belong to this taxon: Buellia minutula (Körb.) Arn., Flora 53: 215. 1870. Note: Imshaug (1951) suggested that Buellia minutula is synonymous with B. stellulata. Hafellner, however, examinded the specimen seen by Imshaug and annotated it as the type of Karschia minutula (unpublished annotation of a specimen seen in M). Buellia maritima (A. Massal.) Bagl., in A. Massal. Schedul. Critic. 8: 150. 1856.— Catolechia maritima A. Massal., Framm. Lich.: 22. 1855. Note: Scheidegger (1993) included B. maritima (= C. maritima) as synonym, but the type is identical with Buellia subalbula according to Bungartz & Nash (2004a). Thallus (Fig. 36A,B) crustose, thin to moderately thickened, ± continuous, epilithic; areolate; hypothallus conspiciously black, in most specimens strongly developed and growing between the areoles, rarely only surrounding the thallus outline; thallus surface matt and dull to ± shiny, usually white to whitish gray, rarely dark gray, epruinose, 295 phenocorticate; without crystals in thallus medulla (H2SO4–). Apothecia lecideine; (0.2–)0.3–0.4(–0.5) mm in diameter; immersed to adnate, rarely sessile; proper margin thin, ± persistent, rarely excluded with age, black or color masked by grayish remains of necrotic thalline material (thalline veil); disc black, epruinose, plane, rarely becoming slightly convex with age; exciple narrow, poorly differentiated, of the aethalea-type (Fig. 36C) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), often reduced, similar in structure and orientation to the paraphyses, transient with the deep reddish brown hypothecium (leptoclinoides-brown, textura intricata), outer excipular hyphae parallel, moderately swollen (textura oblita) and usually strongly carbonized with various amounts of brown and aeruginose pigments (cf. elachista-brown & cinereorufa-green), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachistabrown) and a diffuse, aeruginose pigment (HNO3+ violet, cinereorufa-green). Asci 8spored, clavate, Bacidia-type (Fig. 36D). Ascospores (Fig. 36E,F) oblong to ellipsoid, usually not constricted, with obtuse ends, not curved, (8.0–)8.7–[9.9]–11.1(–13.0) x (4.5–)4.8–[5.5]–6.1(–7.0) µm (n = 60); one-septate, proper septum narrow not thickened during spore ontogeny, lateral wall thickenings absent [Beltraminea (=Buellia)-type]; ornamentation microrugulate, or absent in some specimens; septum with septal pore canal, simple pore and undifferentiated pore plug; spore wall (Fig. 36G) differentiated into smooth, thin perispore (< 0.06 µm), indistinct intermediate layer (< 0.01 µm), thick proper spore wall (0.53–0.68 µm) and moderately thickened endospore (0.12–0.26 µm). 296 → FIGURE 36. Buellia stellulata [A–C. light micrographs (Ryan 31282a), D–E. TEM micrographs (Nash 38464)].—A. Areolate thallus with immersed to slightly sessile, circular apothecia.—B. Close-up of areoles with ± immersed apothecia, some with a thalline veil (arrows).—C. Cross section of an apothecium.—D. Immature Bacidia-type ascus (for designation of the different layers see Bellemère 1994): spores have not yet formed within the ascoplasm (ap); the ocular chamber (oc) is tear-drop shaped; the innermost layer of the tholus (d) is layered but not yet distinctly differentiated into a d1and d2-layer; the c-layer (c) is thick and forms the main ascus wall; a- and b-layer (ab) are barely visible and cannot be reliably distinguished (possibly as a result of fixation artefacts).—E. Ascus with mature ascospores.—F. Mature ascospore.—G. Spore wall of a mature ascospore: (as) ascus wall; (1) perispore; (2) intermediate layer; (3) proper spore wall; (4) endospore. 297 298 Pycnidia rare, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis (1980) and Vobis & Hawksworth (1981); conidia simple, bacilliform, 3.5–4.0 x 0.5–1.0 µm (n = 20). Chemistry.— The depsides atranorin and biosynthetically related 2'-Omethylperlatolic and/or confluentic acid are characteristic for this species. The thalli typically react K+ yellow (sometimes weakly), P– or P+ faintly yellow, C–, KC–, CK–. UV– (dark). The thallus (cortex and medulla) is not amyloid but the apothecia react with Lugol’s. It is also possible to confuse the black hypothallus with a positive iodine reaction when testing the medulla (if testing directly on the specimen concentrated Lugol's iodine should be used). Substrate and ecology.—Growing on hard siliceous rock (generally HCl–), especially near the coast. Distribution (Fig. 10).—Scheidegger (1993, p. 357) suggests that the species occurs “On sun exposed, calcareous or siliceous rocks in southern or central Europe”. The type specimen was, however, collected at an oceanic hillsite, in Southwestern Ireland, not central or southwestern Europe. According to the British Lichen Flora (Orange et al. 1992, p. 136) the species is most typical “On hard, rocks and pebbles, mainly maritime in the xeric supralittoral. N. & W. Britain, Ireland. N. & C. Europe.” Sheard (1964) also emphasizes the coastal distribution of the species. It is very unlikely that the species extends far inland into southern or central Europe and Scheidegger’s (1993) reference 299 thus appears somewhat misleading. Reports of B. stellulata from inland localities should carefully be re-examined. It is possible that these specimens have been misidentified. In the Sonoran Region only specimens from coastal areas of California and Baja California are currently known. Specimens from inland localities of the Sonoran Region identified as B. stellulata mostly belong to B. spuria. Along the coast specimens of B. stellulata have also been confused with B. fimbriata (Tuck.) Sheard or B. cerrusata Llimona & Werner. Both names are synonyms of B. tesserata Körb. (Rico et al. 2003). Buellia stellulata is superficially similar to the maritime B. tesserata but the two species can reliably be distinguished by their distinctly different chemistry (B. stellulata: 2-O’methylperlatolic vs. B. tesserata: divaricatic acid and related substances). Buellia tesserata is also characterized by a more distinctly coarse spore ornamentation (rugulate rather than microrugulate) and the absence of the exciple pigment cinereorufa-green from its apothecia. Buellia stellulata has a dense, black hypothallus forming a distinct thallus outline often extending between the areoles. Buellia tesserata has a fimbriate prothallus, usually not extending between the areoles (i.e. not forming a hypothallus). Additional specimens examined.–CHILE. Juan Fernandez Islands. Imshaug 38014. MEXICO. BAJA CALIFORNIA. Nash 26116, 34217, Nash 12957, Nash 12780b (ASU); Kalb 24673b (hb. Kalb); Weber L-43010, Weber L-43054 (COLO). BAJA CALIFORNIA SUR. Nash 39773 (ASU). NEW ZEALAND. SOUTH ISLAND. Marlborough Co. Nash 19784b (ASU). U.S.A. CALIFORNIA. Del Norte Co. Nash 11532 (ASU). CALIFORNIA. Los Angeles Co. Weber L-42106, Weber L-42733 (COLO); Nash 32099 (ASU). Marin Co. Weber 8159 300 (COLO). Marin Co. Weber 8097 (COLO). San Luis Obispo Co. Riefner L-86315 (COLO). San Mateo Co. Ryan 21975, Shushan S14685 (ASU). Santa Barbara Co. Nash 32343a, 32396, 32748, Ryan 31283 (ASU); Bratt 5385 (SBBG). Santa Cruz Co. Herre L78271 (COLO). Additional species discussed here: BUELLIA LEPIDASTRA (Tuck.) Tuck., Syn. N. Americ. Lich. 2: 90. 1888. Lecidea lepidastra Tuck., Am. Journ. Sci., ser. 2, 25: 249. 1858. TYPE: U.S.A. VERMONT. Windham Co. Brattleborough. On granite, Frost s.n. (FH! – lectotype selected here, two isolectotypes; MICH!). Note: Three type specimens collected by Frost in Vermont are mounted together with several other collection on a sheet at FH with the number 3276. The specimen in the lower right corner has a red sticker with the word “type”. The protologue does not specify a specimen as the holotype and the specimen labeled “type” is selected here as the lectotype. A specimen in M (M-0023904!) is not part of the type but erroneously labeled as such. The original label reads “Buellia lepidastra Tuck., Willey, Massachusetts”. Both the collector “Willey” as well as the locality “Massachusetts” have been crossed out and annotated in a different handwriting as: “Lecidea lepidastra Tuck., Americ. J. Arts and Sci. ser. 2, 25: 429 (1888). Vermont: Brattleborough, Mr. Frost [Windham County] 42°51'03"N, 72°33'30"W.” It is very unlikely that Frost or Tuckerman would have erroneously labeled this collection and later changed the label data. It is much more likely that the original data are correct and the specimen collected by Willey is not part of the 301 type collection. The specimens collected by Willey were mentioned by Tuckerman when he transferred the species into Buellia but only the collection by Frost is mentioned in the protologue of Lecidea lepidastra Tuck. This specimen, not the Willey collection, must therefore be selected as the lectotype. Additional specimens examined.—U.S.A. ALABAMA. County unknown. Peters s.n. (FH). ILLINOIS. Jackson Co. Hatcher 63 (MSC-146975). MASSACHUSETTS. Norfolk Co. Willey s.n. (BM-000660157); Willey 572, 660 (FH). Hampshire Co. Tuckerman s.n. (FH). SOUTH CAROLINA. Chester Co. Green 106 (M-0061352). BUELLIA MACULATA Bungartz, nomen novum pro Buellia stigmaea Tuck., Syn. N. Americ. Lich. 2: 90. 1888. non Körb., Syst. Lich. German.: 226. 1855. TYPE: U.S.A. PENNSYLVANIA. Chester Co. Michener 209 ex Tuckerman sheet 3281 (FH! – lectotype selected here, US! - isolectotype). Note: The species name B. stigmaea Tuck. is illegitimate because it must be considered a later homonym of B. stigmatea Körb. due to the extremely similar spelling of the species epithet: “stigmaea” vs. “stigmatea” (Art. 53.3. ICNB, Greuter et al. 2000). The original epithet “stigmaea” literally means “with stains”. The new epithet “maculata”, meaning “speckled”, has been chosen here to refer to Tuckerman’s original name. The protologue mentions specimens from several localities and a lectotype is therefore selected here. Additional specimens examined.—U.S.A. ILLINOIS. Union Co. Hatcher 47 (MSC0005815). MARYLAND. Baltimore Co. Plitt 28, Plitt 400a (MSC-0005812, MSC-0005813). MASSACHUSETTS. Barnstable Co. Brodo 4203, 4416b, 4447 (MSC-0005806, MSC- 302 0005807, MSC-0005808). NEW YORK. Suffolk Co. Brodo 2672, 3079, 3881 (MSC0005811, MSC-0005810, MSC-0005809). NORTH CAROLINA. Stanley Co. Culberson 5054 (MSC-0005816). TENNESSEE. Hamilton Co. Calkins s.n. (MSC-0005814). KEY TO THE SPECIES 1 Young spores with a distinct septum thickening (Physconia-type); layers below the hymenium initially clearly differentiated into a hyaline subhymenium and a deep reddish brown hypothecium, hypothecium pigmentation later extending and differentiation less visible ............................................................ Buellia lacteoidea Young spores without a distinct median thickening (Beltraminea-type); layers below the hymenium not differentiated, hyaline or pigmented throughout ............ 2 2(1) Thallus without atranorin; apothecia remaining immersed, disc irregular .............. 3 Thallus with atranorin; apothecia emergent, disc circular ...................................... 4 3(2) Thallus thin, pale gray to dark gray, K+ orange to red (with norstictic acid); apothecia predominantly forming in the center of the areoles; exciple and epihymenium aeruginose (cinereorufa-green, HNO3+ violet) .................................. ........................................................................................................ Buellia aethalea Thallus thick, pale gray to pale beige, K–, KC+ pink and C+ pink (with lecanoric acid); apothecia predominantly forming along the margins of the areoles; exciple and epihymenium brown, not aeruginose (HNO3–) ........................... Buellia eganii 4(3) Apothecia with aeruginose, HNO3+ violet pigment (present at least in the outer exciple) ................................................................................................................... 5 303 Apothecia without aeruginose pigment; with a distinct eastern North American distribution ..................................................................................... Buellia maculata 5(4) Thallus granular areolate to verrucose; hypothallus usually faint or absent; apothecia without thalline veil; aeruginose pigmentation usually confined to the outermost exciple; species present only in the North American Northeast (not in the Sonoran Region) .................................................................... Buellia lepidastra Thallus areolate, not granular or verrucose; hypothallus usually distinct, rarely faint or absent; apothecia often with thalline veil; aeruginose pigment extending from the exciple across the epihymenium .............................................................................. 6 6(5) Thallus with norstictic acid (K+ orange to red, forming orange needles in the compound microscope); medulla I+ blue; inland, usually montane .......................... ............................................................................................................ Buellia spuria Thallus without norstictic acid (K–, not forming crystals), instead with 2'-Omethylperlatolic acid and/or confluentic acid; coastal but not necessarily maritime . ........................................................................................................ Buellia stellulata 304 TABLE 6. Key characters of the Buellia aethalea-group from the Greater Sonoran Desert Region and similar North American species. thallus hypothallus apothecia exciple spores medulla (I-/I+) chemistry Buellia aethalea areolate, thin, pale gray to dark gray usually present, rarely faint or absent immersed, central, not becoming sessile, disc irregular aethalea-type cinereorufagreen present Beltraminiatype, i.e. no septum thickening I+ blue or I–; only norstictic acid, no atranorin Buellia spuria areolate, thin to ± thick; whitish gray to dark gray often present and strongly developed emergent, disc circular, frequently with thalline veil aethalea-type cinereorufagreen present Beltraminiatype, i.e. no septum thickening I+ blue; atranorin and norstictic acid Buellia stellulata areolate, thin to ± thick; whitish gray to dark gray often present and strongly developed emergent, disc circular, frequently with thalline veil aethalea-type cinereorufagreen present Beltraminiatype, i.e. no septum thickening I–; atranorin and 2'-O-methylperlatolic and/or confluentic acid SONORAN but possibly not closely related to the Buellia aethalea-group Buellia eganii areolate, thick, pale gray to pale brown often present, but sometimes faint or absent immersed, lateral, not becoming sessile, disc irregular aethalea-type cinereorufagreen absent Beltraminiatype, i.e. no septum thickening I–; lecanoric acid, no atranorin Buellia lacteoidea areolate, thin to thick; ivory to pale creamy white, rarely grayish often present and usually strongly developed immersed, rarely emergent, disc irregular, usually without thalline veil aethalea-type cinereorufagreen present Physconia-type, i.e with septum thickening strongly I+ blue; atranorin, norstictic, rarely gyrophoric acid NOT present in the Southwest Buellia lepidastra granular areolate to verrucose, thin to ± thick; dull sordid gray usually present but not strongly developed emergent, disc circular, without thalline veil aethalea-type cinereorufagreen in outer exciple only Beltraminiatype, i.e. no septum thickening I–; atranorin and norstictic acid Buellia maculata areolate, thin to ± thick; ivory to pale creamy white, rarely grayish often present and strongly developed emergent, disc circular, frequently with thalline veil aethalea-type cinereorufagreen absent Beltraminiatype, i.e. no septum thickening I+ blue,; atranorin and norstictic acid 305 DISCUSSION The Buellia aethalea-group can informally be recognized by a distinctly areolate thallus, often delimited by an extensive black hypothallus, evenly thin-walled Buellia(=Beltraminea)-type ascospores, a Bacidia-type ascus, and lecideine apothecia sometimes remaining immersed in the thallus. These apothecia are characterized by a thin and often reduced exciple colored by the aeruginose HNO3+ violet pigment cinereorufagreen and frequently have a thalline veil. Thallus.—All species discussed here have a distinctly areolate thallus, i.e. with segregated areoles derived from an independent, individual development. Buellia lepidastra is the only species with a ± granular areolate to verrucose thallus. Thalli of all species usually establish on a distinct, black hypothallus, which frequently not only surrounds the entire thallus as a distinct outline but can also delimit separate areoles from one another. The development of this hypothallus varies considerably among specimens of all species and cannot be used to distinguish different species. In a few specimens the black hypothallus is even absent or only poorly developed but in most specimens it is quite distinct. Thallus color of all species varies from an almost pure white to a dark gray. Buellia eganii often has a slight brownish tinge or may even appear distinctly brown. Buellia lacteoidea most typically has a creamy white or ivory thallus. Because of its aeruginose outer exciple and the thin walled ascospores without median thickening, B. lepidastra may belong to the Buellia aethalea-group. However, the thallus of B. lepidastra is distinctly granular areolate to verrucose and only rarely delimited by 306 an indistinct hypothallus. Too little material has so far been examined to assess its affinities to the Buellia aethalea-group. Apothecia.—All species have very similar apothecia, which remain immersed for much of their development. In B. aethalea and B. eganii apothecia do not emerge from the thallus (Figs. 31A,B and 32A,B). This is also the case for most specimens of B. lacteoidea (Fig. 34A,B) but a few specimens of this species have been found with considerably swollen discs which surmount the thallus surface. In all other species apothecia eventually become adnate to sessile with age. The outline of the apothecial disc is initially circular in most species. In B. aethalea, B. eganii and B. lacteoidea (Figs. 32A,B, 33A,B and 34A,B) the discs can become quite irregular and only rarely remain circular in outline. In B. eganii the apothecial disc often becomes moderately convex, whereas it remains plane to concave in B. aethalea. Apothecia of B. aethalea are typically formed in the center of the areoles, those of B. eganii and B. lacteoidea develop along the edges of the areoles. All species have lecideine apothecia. Species with immersed apothecia have been referred to as cryptolecanorine by Scheidegger (1993) even though these species do not have a thalline exciple. In species where the apothecia emerge from the thallus, necrotic thalline material often remains attached to the margin as a thalline veil (Figs. 35B and 36B). This can be very conspicuous in B. spuria and B. stellulata but is rarely observed in other species with emerging apothecia. The proper exciple of all species is fairly thin and considerably reduced. The inner excipular hyphae are similar to the paraphyses and have swollen end cells which form the outer exciple. These outer cells are usually strongly 307 carbonized with a brown pigment cap (cf. elachista-brown) and are often considerably darkened by a diffuse aeruginose pigment (cinereorufa-green). Scheidegger (1993) referred to this thin, considerably reduced exciple as the aethalea-type. He emphasized that this type is characterized by the presence of pigment A (= cinereorufagreen), but nevertheless assigns some species to this particular exciple type which lack the pigment. Cinereorufa-green is absent from B. eganii and B. maculata, even though these species have structurally similar exciples as do all other species discussed here. It is difficult to assess if only the absence of that particular pigment is enough justification to exclude B. maculata and B. eganii from the Buellia aethalea-group. The amount of cinereorufa-green can vary considerably among specimens or even species. In most species the aeruginose diffuse pigmentation extends far into the exciple and across the epihymenium. However, Imshaug (1951) noticed that the pigment is usually confined to the outermost part of the exciple of B. lepidastra. Among these species, both B. aethalea and B. eganii have a hyaline hypothecium (Fig. 32C, 33C), that is quite unusual within the genus. A hyaline hypothecium is more typical for the genus Rinodina, and Buellia is usually characterized by a dark, pigmented hypothecium (Helms et al. 2003). Giralt & Matzer (1994) argue convincingly, however, that no single character can be used to distinguish the two genera and some variation of hypothecium pigmentation has previously been reported in Buellia. According to Scheidegger (1993), specimens of B. aethalea may have either a hyaline or a pigmented hypothecium. We have not observed strong pigmentation, but some of the specimens show a faint brownish tinge. 308 Imshaug (1951) emphasized that the hypothecium of B. lacteoidea is also unpigmented. This assessment has lead to a lot of misidentifications of specimens of B. lacteoidea as B. spuria or B. stellulata. Buellia lacteoidea indeed has a large unpigmented layer of hyphae directly below the hymenium. This colorless subhymenium (Fig. 34C,F) can be distinguished from a pigmented layer below (Fig. 34F), the hypothecium. With age the pigmentation from the hypothecium becomes more pronounced and may extend partially into the subhymenium. In contrast, B. spuria and B. stellulata both have layers of hyphae below the hymenium, which are strongly pigmented throughout. Thus, a colorless subhymenium cannot be distinguished in these species and like in most other species the term hypothecium loosely applies to both layers. Asci and Ascospores.—All species have Bacidia-type asci with a conical, non-amyloid central tholus and amyloid flanks which merge at the ascus tip (Figs. 35E,H and 36D). This ascus type is the most diagnostic difference to distinguish Buellia from Rinodina. Ascospores of all species apart from B. lacteoidea are thin walled with no septum thickening (Beltraminea- or Buellia-type). Buellia lacteoidea has a distinct septum thickening (Physconia-type) at least during some stage of the spore ontogeny. Spore ornamentation of all species is microrugulate but sometimes appears smooth (not ornamented) in specimens of B. spuria and B. stellulata. Spore ornamentation is a direct result of fractures in the perispore wall layer (Nordin 2000). These fractures can reliably only be observed in the TEM. More material of both B. spuria and B. stellulata needs to be examined with the TEM to assess whether the ornamentation is a result of the spore ontogeny. It is possible that the observed variation defines distinct taxonomic groups, i.e. 309 subspecies or even cryptic species within the two taxa. Differences in spore ornamentation could, however, also be related to the ontogeny of the perispore, i.e. fracturing may occur at a later stage of spore development only. Coniodiomata.–All species have bacilliform conidia formed on similar conidiophores in structurally identical pycnidia. Measurements of the conidia are not sufficiently different to segregate the species. Chemistry.–The chemistry of all species is not complex. Buellia aethalea and B. eganii are the only species which consistently lack the depside atranorin. Buellia aethalea is characterized by the depsidone norstictic acid and the thallus reacts K+ yellow becoming deep red (forming orange crystals if observed in the compound microscope). In contrast thalli of B. eganii react K– (or indistinctly yellow) and C+ pink, KC+ pink. Egan (1971) already observed these fleeting reactions and suggested that gyrophoric or lecanoric acid should be present in the species. HPLC analysis confirms the presence of the lecanoric acid. In addition the depside 5-O-methylhiascic acid was also detected. Some specimens of B. lacteoidea contain gyrophoric acid and also react C+ pink, KC+ pink. It was first assumed that thalli of B. spuria and B. stellulata might not consistently be different in their chemistry and possibly belong to the same species. Thalli of B. spuria, however, consistently have atranorin and norstictic acid whereas B. stellulata is characterized by atranorin and 2'-O-methylperlatolic and/or confluentic acid. These differences in secondary metabolites correlate with differences in distribution and iodine reactions of the thallus. 310 As previously reported by Scheidegger (1993) the iodine reaction of the thallus varies in specimens of B. aethalea and can therefore not be used as a distinct character to distinguish this species from any of the other ones. In all other species the iodine reaction of the thallus is, however, very consistent. Thus, the two similar species B. stellulata (medulla I–) and B. spuria (medulla I+ blue) are clearly distinguished by their different reaction. Distribution.–Imshaug (1951) did not include Buellia aethalea in his monograph of North American species, but suggested that reports might be misidentifications. Although the North American Checklist (Esslinger 1997) lists Buellia aethalea, it is not certain which particular specimens these records are based on. Among the Sonoran specimens only very little material can be assigned to Buellia aethalea s.str. Several herbarium specimens identified as B. aethalea from outside the Sonoran Region also do belong to Buellia aethalea s.str. Instead most of the material proved to be Buellia spuria. Thus, it is difficult to assess how common B. aethalea occurs throughout North America. The species appears confined to higher altitudes and is possibly more frequently encountered at high latitudes. Like B. aethalea, Buellia eganii is currently known from only a few localities. All these localities are subalpine to alpine habitats, usually above the timberline. The species was initially considered a synonym of B. aethalea, but is described here as a distinct species based on the absence of cinereorufa-green from the apothecia, comparatively exuberant thalli with larger apothecia forming along the margin of thallus areoles and the presence of lecanoric instead of norstictic acid. 311 In the Sonoran Desert Region Buellia spuria is clearly the most widely distributed species of the group. Buellia spuria is most common throughout montane habitats, but in southern California and Baja California it is also quite frequently encountered at lower elevations. Nevertheless, it is not a typical desert species and absent from the central Sonoran Desert. Along the coast it is regularly confused with Buellia stellulata, an extremely similar species which maintains a distinctly different thallus chemistry and does not extend far inland. Buellia stellulata, although typical along the coast, is not strictly maritime. Imshaug (1951) reported that the species is restricted to the Pacific coast. The species, however, also occurs along the Atlantic, where it has often been confused with B. maculata. Buellia maculata differs from B. stellulata by its different thallus chemistry and the absence of any aeruginose exciple pigmentation. Buellia maculata has apparently the same chemistry as thalli of Buellia spuria but again can be distinguished from B. spuria because of the absence of cinereorufa-green in the exciple. Although previously reported from the region, Buellia lepidastra does not extend into the North American Southwest. Among the material reported as B. lepidastra from the Sonoran Region no specimens matched material from the Northeast. As Imshaug (1951) suggested, the species indeed appears to be confined to the eastern North America. ACKNOWLEDGMENTS We are grateful to Dr. John Sheard, University of Saskatchewan, Canada and Scott Bates, ASU, Arizona, for reviewing the manuscript. Loans from the following herbaria to ASU are greatly appreciated: H, UPS, M, S, US, hb. Scheidegger, OMA, W, MIN, BM, SBBG, COLO, DUKE, MICH, FH. Dr. Robert S. Egan sent us his type material of B. eganii and thus 312 provided us with the opportunity to describe this species. Dr. John A. Elix, University of Canberra, Australia, analyzed selected specimens with HPLC. James Lendemer, Academy of Natural Sciences, Philadelphia provided valuable advice on the figure plates. The study was supported by a Research Grant in Plant Systematics from the International Association of Plant Taxonomists (IAPT) and National Science Foundation Awards to ASU (DEB-0103738, DEB-9701111). A research visit to MSU by the first author was supported by a National Science Foundation Award (DBI-0237401). LITERATURE CITED BELLEMÈRE, A. 1994. Asci and ascospores in ascomycete systematics. pp. 111-126. In HAWKSWORTH, D. L. (ed.) Ascomycete Systematics. Problems and Perspectives in the Nineties, NATO ASI Series, Series A: Life Sciences, 269. Plenum Press, New York. BUNGARTZ, F. & T. H. NASH III. 2004. Buellia subalbula (Nyl.) Müll. Arg. and B. amabilis de Lesd., two species from North America with one-septate ascospores: A comparison with Buellia ["Diplotomma"] venusta (Körb.) Lettau. pp. 49-66. 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A revision of European saxicolous species of the genus Buellia De Not. and formerly included genera. Lichenologist 25: 315-364. SHEARD, J. W. 1964. The genus Buellia de Notaris in the British Isles. (Excluding section Diploicia (Massal.) Stiz.). Lichenologist 2: 225-262. SHREVE, F. & I. L. WIGGINS. 1964. Vegetation and Flora of the Sonoran Desert. Stanford University Press, Stanford. VOBIS, G. 1980. Bau und Entwicklung der Flechten-Pycnidien und ihrer Conidien. Bibliotheca Lichenologica 14: 1-141. VOBIS, G. & D. L. HAWKSWORTH. 1981. Conidial lichen-forming fungi, pp. 245-273. In COLE, G. T. & B. KENDRICK (eds.), 1: Biology of Conidial Fungi. Academic Press, New York. WHITE, F. J. & P. W. JAMES. 1985. New guide to microchemical techniques for the identification of lichen substances. British Lichen Society Bulletin 57: 1-41. 316 PUBLICATION 7 (The Bryologist, submitted February 2004): The genus Buellia s.l. in the Greater Sonoran Desert Region: saxicolous species with one-septate ascospores containing xanthones FRANK BUNGARTZ AND THOMAS H. NASH III Arizona State University Lichen Herbarium, School of Life Sciences, P. O. Box 87 4601, Tempe, AZ 85287–4601, U.S.A. e-mail: frank.bungartz@asu.edu; tom.nash@asu.edu JOHN A. ELIX Department of Chemistry, The Faculties, Australian National University, Canberra, ACT 0200, Australia. e-mail: john.elix@anu.edu.au Abstract. Six species of saxicolous Buellia s.l. containing xanthones are reported from the Greater Sonoran Desert Region: Buellia concinna, B. halonia, B. mamillana, B. prospersa, B. subaethalea and B. trachyspora. All species have a pale yellow to greenish yellow thallus characterized by the presence of xanthones. Nevertheless, they all show distinct characters and may consequently only be distantly related. Buellia concinna, a taxon previously reported only from Europe, is the valid name for the North American B. semitensis. Buellia subaethalea, first described by Bouly de Lesdain, but subsequently largely ignored, is reported for the first time from the region. Buellia trachyspora, a 317 subtropical species barely reaching the Sonoran Region, is distinguished from B. mamillana. Lichen species with xanthones can usually easily be recognized by their yellowish thallus color, frequently have a C+ conspicuously orange thallus reaction and usually fluoresce yellow or orange in UV light. Xanthones are lichen metabolites derived from acetyl CoA via the acetyl polymalonyl pathway. Unlike many other lichen substances, they are difficult to distinguish with routine thin-layer chromatography (TLC). High performance liquid chromatography (HPLC) is therefore the preferred method of identification. In Europe, Scheidegger & Ruef (1988) pioneered analyzing xanthones from saxicolous species of Buellia. Xanthones are not common in Buellia and it is currently difficult to evaluate whether species containing xanthones belong to Buellia s.str. All species from the Sonoran Region are very well distinguished from one another by distinct morphological, anatomical and ultrastructural characters. Even though some of these species may be closely related, it is unlikely that all species with xanthones share a common ancestry. Segregating these species from Buellia s.l. does not resolve the taxonomy of the species, and it is currently not advisable to erect new genera which are monotypic or contain only a few species. All species are therefore included in Buellia s.l. and detailed descriptions are provided including morphological, anatomical, ultrastructural and chemical characters. 318 METHODS All specimens were examined with light microscopy using hand- and cryosections. Both conventional bright field microscopy (BF) and differential interference contrast (DIC) was used. Selected specimens were also studied with transmission electron microscopy (TEM) according to a protocol described in detail by Bungartz et al. (2002). To improve dehydration and infiltration, this protocol has been modified according to Bungartz & Nash (2004a). All specimens were spot tested and routinely examined with standardized thin-layer chromatography (Culberson & Kristinsson 1970; Culberson & Johnson 1982; White & James 1985; Orange et al. 2001). TLC-plates were interpreted with the computer program WINTABOLITES (Mietzsch et al. 1994), and scanned for permanent record (Egan 2001). In addition a subset of specimens were analyzed using standardized high performance liquid chromatography (HPLC, Elix et al. 2003). Spores measurements are given according to Nordin (2000). Pigment names follow Meyer & Printzen (2000). SPECIES DESCRIPTIONS BUELLIA CONCINNA Th. Fr., Nova Acta Reg. Soc. Scient. Upsal. Ser. 3, 3: 332. 1860. TYPE: NORWAY. FINMARK. Varanger, Nesseby, 30 August 1857, Fries s.n. (UPS – lectotype designated by Foucard et al. 2002, BM, GZU, M!, O, PC – isolectotypes). Note: Zahlbrucker (1931) erroneously lists “1861” instead of 1860 as the publication date. Buellia semitensis Tuck., Syn. N. Amer. Lich. 2: 95. 1888. TYPE: U.S.A. CALIFORNIA. Mariposa Co. Granitic rocks, Bolander 329 ex Tuckerman sheet 3281 (FH–lectotype selected here!). Note: The lectotypefication is necessary because Tuckerman (1888, p. 96) 319 did not refer to a particular specimen in the protologue: “Granitic rocks, Yosemite valley and elsewhere, California, Bolander:” For additional synonyms see Scheidegger & Ruef (1988) and Foucard et al. (2002). Thallus (Fig. 37A,B) crustose, thin to moderately thickened, closely aggregated or dispersed, epilithic; granular to minutely bullate; prothallus absent; thallus surface shiny and smooth, rarely matt, usually pale yellow to greenish yellow, rarely becoming pale yellow (older herbarium specimens), epruinose, phenocorticate; lacking Ca-oxalate crystals within thallus medulla (H2SO4–, no clusters of Ca-sulphate needles forming). Apothecia lecideine; (0.2–)0.3–0.6(–0.8) mm in diameter; soon sessile; proper margin prominent, usually persistent, rarely excluded with age, black; disc black, epruinose or rarely with faint, yellow pruina, plane, rarely becoming slightly convex with age; exciple distinct, of the leptocline-type (Fig. 37C) sensu Scheidegger (1993), i.e. exciple thick, distinct and not distinctly differentiated into an inner and outer part, hyphae thin-walled, prosoplectenchymatous and usually ± densely interwoven (textura intricata), dull fuscous brown throughout, becoming ± carbonized by various amounts of brown pigments (cf. elachista-brown), pigmentation continuous with the epihymenium, hypothecium deep reddish brown (leptoclinoides-brown, textura intricata); hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown (cf. elachista-brown) pigment cap and a diffuse, brown pigment (HNO3+ violet, cinereorufagreen). Asci 8-spored, clavate, Bacidia-type. Ascospores (Fig. 37D–I) ellipsoid to ± crescent-shaped, i.e. often distinctly curved and characteristically with tapered ends, 320 rarely constricted, (13.0–)15.6–[18.5]–21.3(–24.0) x (6.0–)7.9–[8.9]–9.9(–11.0) µm (n = 60); one-septate (very rarely with additional median septa forming at both ends), proper septum becomes thickened early during spore ontogeny (Physconia-type; Fig. 37D,E); ornamentation microrugulate, becoming rugulate with age (best seen in DIC); septum with septal pore canal, simple pore and undifferentiated pore plug; spore wall (Fig. 37G,I) differentiated into cracked, thin perispore (0.11–0.17 µm), narrow intermediate layer (< 0.03 µm), thick proper spore wall (0.37–0.50 µm) and moderately thickened endospore (0.24–0.34 µm). Pycnidia rare, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform to ellipsoid, 2.0–4.0 x 1.5–2.0 µm (n = 20). Chemistry (Fig. 42).—Thallus characterized by the following xanthones: arthothelin, isoarthothelin, 6-O-methylarthothelin, 4,5-dichloronorlichexanthone, 4,5-dichloro-6-Omethylnorlichexanthone, asemone and thiophanic acid. In addition, the monocyclic phenol derivatives gyrophoric, lecanoric and orsellinic acid are often present. Spot test reactions usually C+ orange or sometimes ± pinkish, K+ yellow, KC+ orange, P+ orange, CK+ orange. UV+ pale or bright yellow to orange. The thallus medulla reacts ± amyloid or not amyloid; apothecia react amyloid in Lugol’s (always test with concentrated Lugol's iodine or in the compound microscope; the reactions can be very weak!). 321 Substrate and ecology.—On a variety of hard, siliceous (HCl–) rock substrates, often on vertical cliffs. Scheidegger & Ruef (1988) report that the species often grows as a juvenile parasite on crustose and rarely foliose lichens. The North American specimens are not lichenicolous but otherwise match the description of B. concinna in Scheidegger & Ruef (1988), Scheidegger (1993) and Foucard et al. (2002). Isolectotype material of B. concinna, examined during a visit to M, as well as a specimen from Scheidegger’s private herbarium are not lichenicolous and agree well with the North American material. Distribution (Fig. 7).—Throughout the Sonoran Desert Region moderately common at higher elevations (montane to subalpine). Widely distributed throughout the Northern Hemisphere. Notes.—Buellia concinna is easily recognized by its distinct granular thallus and the very characteristic, crescent-shaped spores. This unusual spore shape is not present in all spores of an apothecium but can usually be found in at least some of the sections examined from an apothecium. No other species from the Sonoran Region has similar spores. Selected specimens examined.—FRANCE. PYRÉNÉES-ORIENTALES. Scheidegger 7840 (hb. Scheidegger). MEXICO. HIDALGO. Nash 38098 (ASU). SONORA. Nash 12483 (ASU). NORWAY. FINMARK. Fries s.n. (MSC-69240). U.S.A. ARIZONA. Apache Co. Nash 27088, 11791 (ASU), Hertel 40189 (M). Cochise Co. Nash 14596, 19499 (ASU); Nordin 5178 (UPS). Coconino Co. Nash 7575 (ASU). Santa Cruz Co. Nash 18578 (ASU). CALIFORNIA. El Dorado Co. Ryan 23591a (ASU). CALIFORNIA. Fresno Co. Thiers 34410 (ASU). Lake Co. Sigal s.n. (ASU). Los Angeles Co. Ryan 26137b (ASU). San Diego Co. 322 Nash 14654, Ryan 25821b (ASU). Shasta Co. Nash 22836, Ryan 11352 (ASU). Tulare Co. Nash 11287 (ASU). Tuolumne Co. Ryan 24231, 24490, 23959 (ASU). COLORADO. Clear Creek Co. Anderson 6673 (ASU). El Paso Co. Anderson 8293 (ASU). SOUTH DAKOTA. Custer Co. Anderson 7879 (ASU). Unknown Co. Anderson s.n. (MSC-63646). NEW ZEALAND. Nash 19094, Nash 19112 (ASU). BUELLIA HALONIA (Ach.) Tuck., Lich. Californ.: 26. 1866. Lecidea halonia Ach., Meth. Lich.: 47. 1803. TYPE: SOUTH AFRICA. CAPE PROVINCE. Cape of Good Hope. On hard maritime rock [original label data: Caput bonae spei, in saxis durissimis ad littora Africae australioris], Osbeck s.n. (H-ACH–362! – holotype & several isotypes; UPS-ACH – isotype); ex herb. Acharius (H-NYL 9211 – possibly a type fragment). Buellia disciformis var. halonia (Ach.) Boist., Nouv. Flore Lich. 2: 235. 1903.— Lecidea disciformis var. halonia (Ach.) Nyl., Mém. Soc. Imp. Sci. Natur. Cherbourg 5: 126. 1857. Note: These combinations are based on the same type as B. halonia. Baeomyces capensis Taylor in Hook., London Journ. Bot. 6: 186. 1847.—Diploicia capensis (Taylor) Dodge, Beih. Nov. Hedwigia 38: 157. TYPE: SOUTH AFRICA. CAPE PROVINCE. Cape of good hope. ex Hooker herb. sub. “9311 Lecidea” in Taylor herb. sheet 1460 (FH – holotype, BM-000660172, BM-000660173 - isotypes). Possible synonyms: Buellia flavoareolata (Nyl.) Müll. Arg., Hedwigia 31: 283. 1892.—Lecidea flaveoareolata Nyl., Annal. Scienc. Nat. Bot., ser. 4, 3: 166. 1855. TYPE: CHILE. 323 QUEBRADA. Quilmenco, 31°45'S, 71°05'W. (H-NYL – syntype). Note: Specimens from Chile (Coquimbo Province) distributed by Weber Lich. Exs. 564a, b (ASU, SBBG), and material collected by Imshaug on an Expedition to Santa Clara (Juan Fernandez Islands, Chile; MSC-0005485, MSC-0005486, MSC-0005487, MSC-0005488) are morphologically and anatomically very similar if not identical to B. halonia. These specimens were analyzed with TLC by R.C. Harris, who detected atranorin and an unknown xanthone. Imshaug annotated several other specimens (see specimens examined) from San Juan Fernandez (Chile) as B. halonia. These specimens were not examined by TLC or HPLC but agree well with B. halonia. The type of B. flavoareolata has not been studied but it is very likely that it is a synonym of B. halonia. Thallus (Fig. 38A,B) crustose, thick, ± continuous, epilithic; areolate; prothallus distinct, delimiting the thallus as a black outline; thallus surface matt and smooth, ± shiny, usually yellowish green to pale yellow, rarely becoming pale yellow (older herbarium specimens), epruinose, phenocorticate; lacking Ca-oxalate crystals within thallus medulla (H2SO4–, no clusters of Ca-sulphate needles forming). Apothecia lecideine; (0.2–)0.3–0.5(–0.7) mm in diameter; initially immersed, young apothecia frequently ± aspicillioid, soon bursting through the thallus surface and becoming adnate to sessile; proper margin prominent, usually persistent, rarely excluded with age, black, frequently with coarse thallus fragments attached; disc black, epruinose, sometimes with yellowish pruina, plane, rarely becoming strongly convex with age; exciple distinct, of the leptocline-type (Fig. 38C, D) sensu Scheidegger (1993), i.e. exciple thick, distinct and 324 not distinctly differentiated into an inner and outer part, hyphae thin-walled, prosoplectenchymatous and usually ± densely interwoven (textura intricata), dull fuscous brown to aeruginose throughout, becoming ± carbonized by various amounts of brown and aeruginose pigments (cf. elachista-brown & cinereorufa-green), pigmentation continuous with the epihymenium, but aeruginose pigment not extending into the deep reddish brown hypothecium (leptoclinoides-brown, textura intricata); hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown) and a diffuse, aeruginose pigment (HNO3+ violet, cinereorufa-green). Asci 8-spored, clavate, Bacidia-type. Ascospores (Fig. 38E–I,L) oblong to ellipsoid, usually not constricted, with obtuse ends, not curved, (11.5–)12.4– [14.1]–15.7(–19.0) x (6.0–)6.8–[7.4]–8.0(–9.0) µm (n = 60); one-septate, proper septum soon but only briefly thickened during the ontogeny (Physconia-type; Fig. 38E,F); ornamentation weakly microrugulate (in DIC); septum with septal pore canal, simple pore and undifferentiated pore plug; spore wall (Fig. 38H,L) differentiated into smooth, thin perispore (0.14–0.20 µm), narrow intermediate layer (< 0.05 µm), thick proper spore wall (0.35–0.44 µm) and moderately thickened endospore (0.18–0.34 µm). Pycnidia (Fig. 38J) rare, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980, Fig. 38K); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 5.0–7.0 x 1.0–1.5 µm (n = 20). 325 Chemistry (Fig. 42).—The following xanthones were found with HPLC: arthothelin, isoarthothelin, 2,4-dichloronorlichexanthone, 2,5-dichloronorlichexanthone, 2,7-dichloronorlichexanthone, 4,5-dichloronorlichexanthone, 5,7-dichloronorlichexanthone, thiophanic and thiophaninic acid. The depside atranorin is also regularly present, frequently with the depsidone norstictic acid. Two bis-xanthones, eumitrin X and eumitrin Y, have been detected with HPLC in part of the specimens. Their chemical structure is currently still unresolved. Thalli typically react K+ yellow, P+ orange, C+ orange, KC+ orange and CK+ orange. Fluorescence is typically UV+ bright yellow to orange. The thallus is not amyloid, but apothecia react amyloid in Lugol’s. Substrate and ecology.—Growing abundantly on maritime siliceous mineral-poor coastal rock (generally HCl–). Distribution (Fig. 8).—Very common along the coast of southern California and Baja California. Currently known from the Pacific coast of North America, South Africa, Australia and Chile (see notes on B. flaveoareolata, which is possibly a synonym of B. halonia). Reports of this species from Europe belong to B. concinna according to Scheidegger & Ruef (1988). Notes.—Buellia halonia has frequently been confused with Lecidella asema (Nyl.) Knoph & Hertel, a crustose lichen with similar color, often growing in the same habitat. In the field granulose thalli of L. asema can usually be distinguished from areolate thalli of B. halonia. This distinctly different morphology is often sufficient for identification but the two species are more reliably distinguished by microscopic examination: 326 Lecidella asema has simple, hyaline spores. Spores of Buellia halonia are brown and one-septate. Selected specimens examined.—AUSTRALIA. SOUTH AUSTRALIA. Streimann 54946 (MSC-354293). CHILE. JUAN FERNANDEZ. Imshaug 37584 (MSC-113121); Imshaug 38241 (MSC-113136); Imshaug 38236 (MSC-113137); Imshaug 38204B (MSC-128905). MEXICO. BAJA CALIFORNIA. Nash 34051, 17166, 34202, 38387, 38565, 12955, 4910, 4911, 4922, 8690, 8719, 25186, Ryan 21332, 21333, 21462(ASU); Kalb 24668a (hb. Kalb); Weber L-36628, L-43100, L-43055 (COLO); Wetmore 63671 (MIN). BAJA CALIFORNIA SUR. Nash 40017 (ASU). SOUTH AFRICA. CAPE PROVINCE. Eaton s.n. (BM); unknown collector (H-NYL 9212); Eaton s.n. (H-NYL 2210). U.S.A. CALIFORNIA. Lake Co. Toren s.n. (SFSU), Hasse s.n. (ASU). Los Angeles Co. Marsh 6500, Nash 32020, Ryan 30942 (ASU); Weber L-42957, L-42176, L-42769, S-996, L-42661, L-42632 (COLO). San Diego Co. Bratt 5628 (SBBG). San Luis Obispo Co. Nash 36976, 36985a, Riefner 88-431 (ASU). San Mateo Co. Shushan S14688 (ASU). Santa Barbara Co. Nash 32573, 32574, 32718, 41246, 41253, 41322, 33102, Ryan 31184, Weber Lich. Exs. 680 (ASU); Weber L80190 (COLO). Ventura Co. Nash 37035, 37088, 38689, 38909 (ASU); Weber S1734 (COLO). BUELLIA MAMILLANA (Tuck.) W. A. Weber, Mycotaxon 27: 493. 1986. Rinodina mamillana Tuck., Proc. Amer. Acad. 7: 226. 1866 (1868). TYPE: U.S.A. HAWAII. Honolulu. Oahu. Volcanic Rocks, Mann s.n. ex sheet no. 2124a (FH–holotype, W–isotype!). 327 Buellia glaziouana (Kremp.) Müll. Arg., Flora 63: 19. 1880.—Lecidea glaziouana Kremp., Flora 59: 317. 1876. TYPE: BRAZIL. RIO DE JANEIRO. On granitic rocks [original label data: ad saxa granitica], Glaziou 3506 (M-0023763! – holotype, M0023765!, BM-000660175! – isotypes, C!, G! W!, – isotypes). Buellia thomae (Tuck.) Imshaug comb. ined., in Imshaug University Microfilms no. 2607. 1951.—Rinodina thomae Tuck., Syn. N. Amaric. Lich. 1: 209. 1882. TYPE: U.S.A. ALABAMA. Lawrence Co. Sandstone Rocks at Moulton, 34°28'N 87°17'W, ca. 190 m, Peters s.n. (= ex Tuckerman sheet no. 2125) (FH – holotype, MICH – isotype). Melanaspicilia contiguella (Vain.) Vain., Ann. Acad. Sci. Fenn. ser. A 6: 79. 1915— Buellia contiguella (Vain.) Malme, Bih. K. Svenska Vet.-Akad. Handl. 28: 5. 1902.— Rinodina contiguella Vain., Étude Lich. Brésil 1: 164. 1890. TYPE: BRAZIL. RIO DE JANEIRO. On granitic rock [original label data: ad saxa granitica], Glaziou 3506 (M! – holotype, isotypes). Buellia moreliiensis de Lesd., Lich. Mex: 26. 1914. TYPE: MEXICO. MICHOACÁN. Morelia. On volcanic rocks (US! – lectotype selected here). Note: The material from US is selected here as the lectotype because the holotype material was probably destroyed in World War II. Imshaug (1955) reported that he had seen type material from Magnusson’s private herbarium. Buellia glaziouana var. poliocheila (Vain.) Imshaug, Farlowia 4: 493. 1955.—Buellia poliocheila Vain., Ann. Acad, Sci. Fenn. ser. A 6(7): 85. 1915. TYPE: U.S.A. VIRGIN ISLANDS. St. Thomas. On rocks at Ma Folie, Biese s.n. (C–holotype, TUR–isotype). 328 Buellia recepta (Kremp.) Müll. Arg., Linnaea 43: 36. 1880.—Lecidea recepta Kremp., Flora 59: 318. 1876. TYPE: BRAZIL. RIO DE JANEIRO. On granitic rock. [original label data: ad saxa granitica], 1869, Glaziou 3295 (M-0023811–holotype!, M-0023810– isotype!). Buellia glaziouana var. sensitiva (Zahlbr.) Imshaug, Farlowia 4: 494. 1955.—Buellia sensitiva Zahlbr., Mycologia 22: 78. 1930. TYPE: U.S.A. PUERTO RICO. Naranjito. In an open field, 25 November 1915, Fink 102 (MICH–holotype!). Buellia antillarum de Lesd., Bull. Museum Hist. Nat. Paris II 15: 470. 1934. TYPE: FRENCH WEST INDIES. GUADELOUPE ISLAND. Cove south of Bouillante. On volcanic rock at sea level. [original label data: Anse du Sud de Bouillante] collector not known (PC–holotype). Buellia cohibilis (Nyl.) Zahlbr., Cat. lich. univ. 7: 344. 1931.—Lecidea cohibilis Nyl., Sertum ich. Trop. Labuan et Singapore 41. 1891. TYPE: U.S.A. TENNESSE. At Lookout Tower mountain, on sandstone, Calkins s.n. (H-NYL – holotype; F, FH, MICH, MO, NY, US– isotypes). Possible synonym: Buellia subglaziouana S.R. Singh & D.D. Awasthi, Biological Memoirs 6: 190-191. 1981. TYPE: INDIA. MADHYA PRADESH. Hoshangabad district, Pachmarhi, on way to Chhota Mahadeo, altitude ca. 1050 m, 26 January 1973, Singh 73.157 (AWAS – holotype). Note: No type material of B. subglaziouana has been studied. Singh & Awasthi (1981) described B. subglaziouana as different from B. glaziouana because of a deep iodine-blue medulla reaction. However, contrary to the protologue B. glaziouana also has a distinctly 329 I+ blue medulla. Buellia subglaziouana could therefore be considered a synonym of B. mamillana (= B. glaziouana). Thallus (Fig. 39A,B) crustose, thin to moderately thick, ± continuous, epilithic; rimose, rarely becoming rimose-areolate; prothallus distinct, delimiting the thallus as a black outline; thallus surface matt and smooth, rarely ± shiny, usually pale greenish yellow, rarely gray (older herbarium specimens), epruinose, phenocorticate; lacking Caoxalate crystals within thallus medulla (H2SO4–). Apothecia of the mamillana-type, i.e. initially lecanorine, but soon becoming lecideine; (0.3–)0.6–0.9(–1.1) mm in diameter; usually adnate to sessile, rarely remaining immersed; proper margin thickening with maturity, inconspicuous in immature apothecia, but soon becoming distinct, brownish black; disc dark brown to blackened, epruinose, plane, usually ± convex with age; exciple of immature apothecia thin, indistinct but soon thickening, i.e. inner excipular hyphae distinct, hyaline but becoming pigmented with age, prosoplectenchymatous (textura angularis), distinct, not reduced, similar in structure and orientation to the deep reddish brown hypothecium (leptoclinoides-brown, textura intricata), outer excipular hyphae short-celled, cells angular, ± swollen (textura angularis) and usually ± carbonized with various amounts of brown pigments (cf. elachista-brown), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown (cf. elachista-brown) pigment cap. Asci 8spored, clavate, Bacidia-type. Ascospores (Fig. 39E–G) oblong to ellipsoid, usually not constricted, with obtuse ends, not curved, (10.5–)12.0–[13.8]–15.7(–18.5) x (5.5–)6.3– 330 [7.0]–7.8(–10.0) µm (n = 60); one-septate, proper septum soon but only briefly thickened during spore ontogeny (± Physconia-type); ornamentation rugulate; septum with septal pore canal, simple pore and undifferentiated pore plug; spore wall (Fig. 39G) differentiated into smooth to cracked, thick perispore (0.32–0.43 µm), indistinct intermediate layer (< 0.01 µm), thin proper spore wall (0.09–0.16 µm) and moderately thickened endospore (0.17–0.28 µm). Pycnidia rare, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. coniophore-type III sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, fusiform, 6.0–14.0 x 1.0–1.5 µm (n = 20). Chemistry (Fig. 43).—The following xanthones where detected with HPLC: 4,5dichloro-3-O-methylnorlichexanthone, 4,5-dichlorolichexanthone, 4-chlorolichexanthone. The depside atranorin is also present, often accompanied by haematommic acid and methyl-β-orsellinate, both artifacts formed by hydrolysis of atranorin. 2'-OMethylperlatolic, divaricatic acid, norstictic, hypostictic, stictic, methylstictic, and cryptostictic acid and one unknown trace substance have also been detected by HPLC. Thalli usually react K+ yellow, P– or P+ yellow, C–, KC–, CK–, but these spot tests are sometimes weak. Thalli generally fluoresce UV+ bright or pale yellow to orange. Both thallus medulla and apothecia react distinctly amyloid. Imshaug (1955) first noticed an amyloid medulla reaction contrary to Vainio (1915) and Malme (1902), who reported a negative iodine reaction. Weber (1986) confirmed Imshaug’s observation of the positive iodine reaction. 331 Substrate and ecology.—On shaded, steep, ± vertical, siliceous rock faces (HCl–) Distribution (Fig. 8).—A montane subtropical species, common in the Sierra Madre Occidental and Southern Baja California. Known from tropical to subtropical localities throughout the world (Southeastern USA: Tuckerman 1888; Brasil: Malme 1902; Mexico: Bouly de Lesdain 1914; Caribbean: Imshaug 1955; 1957; India: Singh & Awasthi 1981). Notes.—Based on the development of the apothecia Imshaug (1955) distinguished three lines of development in B. mamillana and recognized two of these lines as varieties: (1) with apothecia remaining immersed, (2) with apothecial initials frequently abortive (var. poliocheila) and (3) with a depsidone and a K+ red spot test reaction, forming crystals (var. sensitiva). This range of variation can be explained if apothecial anatomy is examined carefully. All apothecia are initially immersed but eventually emerge from the thallus with a distinct thalline margin. Immature apothecia do not appear abortive because spores are frequently formed even in young apothecia, which are not yet enclosed by a distinctly colored envelope. The considerable variation in thallus chemistry, as confirmed by HPLC, would suggests that Imshaug’s varieties might correlate with distinct chemotypes and if so, these varieties would deserve separate taxonomic recognition. Material examined here could, however, not be segregated into distinct chemotypes. It was not possible to correlate the chemical variation with any of the “developmental lines” described by Imshaug (1955), therefore no distinct varieties are recognized here. 332 Selected specimens examined.—ARGENTINA. TUCUMAN. Lamb 5164 (MSC117950). BRAZIL. RIO DE JANEIRO. Vainio Lich. Bras. Exs. 74 (M-0061345, BM000671862); Glaziou s.n. (MSC-122653); Lamb 5014 (MSC-338305). SÃO PAULO. Kalb Lich. Neotrop. Exs. 46 (M-0061341); Lamb 5013 (MSC-117955). BRITISH ANTILLES. ST. LUCIA. Imshaug 30160, 30160, 29661, 29661, 29649, 29649, 30130, 30130, 30130, 30130 (MSC-347926, MSC-347926, MSC-76213, MSC-76213, MSC-76212, MSC-76212, MSC-347927, MSC-347927, MSC-348114, MSC-348114). CHILE. ISLA DE PASCUA (= EASTER ISLAND). Skottsberg s.n. (BM-000671861). COSTA RICA. Pittier s.n. (BM000660174, BM-000671861). CUBA. LA HABANA. Imshaug 24815 (MSC-75987). DOMINICAN REPUBLIC. DE LA VEGA. Imshaug 23738 (MSC-76115); Wetmore 23738 (MSC-76115). ECUADOR. GALAPAGOS ISLANDS. Weber Lich. Exs. 110 (M-0061342); Hill s.n. ex sheet no. 2124a (FH). FRENCH ANTILLES. GUADELOUPE. LeGallo 2707 (MSC-60972). SAINT-BARTHÉLEMY. LeGallo 2665 (MSC-61050); LeGallo 2655 (MSC61038); LeGallo 2660, 2650, 2656, 2658, 2670, 2670, 2577, 2556 (MSC-61047, MSC61071, MSC-61051, MSC-61048, MSC-61043, MSC-61044, MSC-61074, MSC-61049). JAMAICA. ST. ANDREW PARISH. Imshaug 14454, 13808, 13808, 13815, 14005, 13985, 15038 (MSC-76027, MSC-76062, MSC-148458, MSC-148457, MSC-76065, MSC-76064, MSC-148459). MEXICO. BAJA CALIFORNIA. Ryan 21461 (ASU). BAJA CALIFORNIA SUR. Nash 29793, 33778, 16953, 1262, 12640, 12743, 39733, 39923 (ASU). CHIHUAHUA. Nash 31295, 13534, 13725 (ASU). NAYARIT. Nash 39192 (ASU). SINALOA. Nash 10086, 12163a, 12214 (ASU). SONORA. Marsh 5918, Nash 25518, 25604, 30352, 33611, 10955, 11008, 11111, 11130, 11132, 11942, 11958, 11993, 12031, 12335, 1244, 14769, 10959d, 333 10964d, 12040b, 10974b, 25524, Ryan 21625, 21636, 21637, 21691a (ASU); Wetmore 69963 (COLO). PARAGUAY. DEPARTMENTO DE PARAGUARI. Malme 1486B (MSC129597). SAINT VINCENT & THE GRENADINES. SAINT VINCENT. Imshaug 30568, 30568 (MSC-347923, MSC-347923). U.S.A. ARIZONA. Cochise Co. Nash 9653b, Ryan 10704 (ASU). Gila Co. Nash 28439. Pima Co. Nash 7984 (ASU). Santa Cruz Co. Nash 25875, 13004, 20698 (ASU). ARKANSAS. Garland Co. Wetmore 86251 (MSC-379125). HAWAIIAN ISLANDS. Kauai Nash 20829 (ASU); Hawaii Co. Bailey 353 (DUKE). LOUSIANA. Natchitoches Co. Tucker 17523 (M-0061343). TENNESSEE. Hamilton Co. Calkins 224, 236 (MSC-146694, MSC-120013). TEXAS. Nacogdoches Co. Wetmore 17638 (MIN). BUELLIA PROSPERSA (Nyl.) Riddle, Brookl. Bot. Gardens Memoirs 1: 114. 1918. Recent synonyms: Rinodina lecideina H. Mayrhofer & Poelt, Bibl. Lich. 12: 112 (1979). Amandinea lecideina (H. Mayrhofer & Poelt) Scheid. & H. Mayrhofer, in Scheidegger, Lichenologist 25: 342 (1993) For a detailed description see Bungartz et al. (2004b). BUELLIA SUBAETHALEA de Lesd., Lichens du Mexique: 27. 1914. TYPE: MEXICO. MICHOACÁN. Morelia, Santa Maria mountain ridge [original label data: Loma Santa Maria] 2000 m, Frère Arsène Brouard 3654 (MSC 48484–lectotype selected here!; US!, S!–isolectotypes). Note: The lectotype is selected here because the holotype in Dunkerque was probably destroyed in World War II. 334 Thallus (Fig. 40A,B) crustose, thin to moderately thickened, ± continuous, epilithic; granular-areolate to verrucose; prothallus absent or delimiting the thallus as a black outline; thallus surface matt and smooth, ± shiny, usually pale yellow, rarely becoming yellowish gray (in the herbarium), epruinose, phenocorticate; lacking crystals within thallus medulla (H2SO4–). Apothecia lecideine; (0.1–)0.2–0.4(–0.6) mm in diameter; immersed, becoming adnate to sessile; proper margin thin, reduced, inconspicuous, black; disc black, epruinose, plane, rarely becoming slightly convex with age; exciple narrow, poorly differentiated, of the aethalea-type (Fig. 40C) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), often reduced, similar in structure and orientation to the paraphyses, transient with the deep reddish brown hypothecium (leptoclinoides-brown, textura intricata), outer excipular hyphae parallel, moderately swollen (textura oblita) and usually strongly carbonized with various amounts of brown pigments (cf. elachista-brown, HNO3–), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown (cf. elachista-brown) pigment cap. Asci 8spored, clavate, Bacidia-type. Ascospores (Fig. 40D–I) oblong to ellipsoid, often constricted, with obtuse ends, not curved, (12.0–)14.2–[16.4]–18.6(–22.0) x (6.0–)7.1– [8.1]–9.0(–11.0) µm (n = 60); one-septate, proper septum developing soon and rarely thickened during spore ontogeny, with two dark bands across each cell (Fig. 40D; ± Physconia-type); ornamentation rugulate; septum with septal pore canal, simple pore and undifferentiated pore plug; spore wall (Fig. 40G,H) differentiated into fractured, thick 335 perispore (0.31–0.47 µm), narrow intermediate layer (< 0.04 µm), moderately thickened proper spore wall (0.17–0.29 µm) and moderately thickened endospore (0.12– 0.22 µm). Pycnidia rare, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 7.0–11.5 x 1.0–1.5 µm (n = 20). Chemistry (Fig. 43).— The only xanthone detected in this species is 4,5dichlorolichexanthone. In addition the biosynthetically related depsidones norstictic, cryptostictic, methylstictic and stictic acid were detected. The thallus reacts K+ yellow to orange (forming orange needle-shaped crystals in the compound microscope), P+ yellow to orange, C–, KC–, and CK–. Thalli show a dull fluorescence, i.e. UV+ deep orange. The thallus is not amyloid, but apothecia react amyloid in Lugol’s (always test with concentrated Lugol's iodine or in the compound microscope; positive reactions can be very weak!). Substrate and ecology.—On a variety of siliceous (HCl–) rock substrates. Distribution (Fig. 10).—A montane, subtropical species, infrequent in southeastern Arizona and probably more common towards the Sierra Madre Occidental. Originally described from Michoacán, Mexico but little is presently known about the extent of the distribution. 336 Notes.—Buellia subaethalea is superficially similar to B. concinna but the thallus is verrucose-areolate rather than granular and the oblong to ellipsoid spores are never crescent-shaped and usually distinctly “banded” (see Discussion – Asci and Ascospores). Selected specimens examined.–MEXICO. CHIHUAHUA. Nash 31382 (ASU). DURANGO. Nash 31087 (ASU). SINALOA. Nash 10147, 12167 (ASU). SONORA. Nash 10871 (ASU). U.S.A. ARIZONA. Apache Co. Nash 26815, 27024, 27028, 27029 (ASU). Cochise Co. Ryan 10748, 10771, 10991 (ASU); Weber S28018, L-36928 (COLO). Gila Co. Nash 39439, 39476 (ASU). Pima Co. Darrow 722, Nash 21208, 38955 (ASU). Santa Cruz Co. Nash 25353, 18554, 8292 (ASU). TEXAS. Brewster Co. Nash 15037b (ASU). BUELLIA TRACHYSPORA Vain., Annal. Acad. Sc. Fennic. ser. A 6(7): 84. 1915. TYPE: U.S.A. VIRGIN ISLANDS. St. John Co. Reef Bay. On a cliff along a cataract [original label data. Indias occ. S. Jan, Reef Bay, In rupe ad cataractam], 7 March 1906, Boergesen 19067-3 (= Vainio 09521a) (C! – lectotype; TUR-10017! – isolectotype, both selected by Imshaug 1955). Note: The former Danish Virgin Island “St. Jan” is now part of the U.S.A. and more frequently referred to by its English name “St. John”. Imshaug (1955) regarded material in C as the holotype because most of Vainio’s tropical material is deposited there. However, the protologue does not mention a particular herbarium (TUR or C), where the holotype is deposited. Imshaug’s annotation thus must be interpreted as a lectotypification (see Art. 10.5 ICBN). The material in TUR is thus isolectotype material. Only a small fragment of this isolectotype material (Boergesen 9521a) was made available for study at ASU. This fragment is annotated as “only part of the type (# 9521) 337 S. Huhtinen, Curator, Turku University Herbarium (TUR)”. The lectotype material at C is a well developed, large specimen and annotated in Vainio’s handwriting as “type”. Buellia gyrosa Vain., Annal. Acad. Sc. Fennic. ser A. 6(7): 85. 1915. TYPE: U.S.A. VIRGIN ISLANDS. St. John Co. On rock. 15 March 1906, Boergesen s.n. (Vainio 9583; C! – holotype, TUR – isotype). Note: Imshaug (1955) studied type material from C and TUR and argued that B. gyrosa “…differs from B. trachyspora only in the gyrose nature of the apothecia…” (p. 505). The older apothecia of the type specimen have a folded margin. We agree with Imshaug’s (1955) assessment that this variation “…scarcely warrants species recognition” (p. 505). Thallus (Fig. 41A,B) crustose, thin, ± continuous, epilithic; rimose to rimose-areolate; prothallus distinct, delimiting the thallus as a black outline; thallus surface matt and smooth, not shiny, usually pale greenish yellow, rarely gray (herbarium specimens), epruinose, phenocorticate; lacking crystals within thallus medulla (H2SO4–). Apothecia lecideine; (0.3–)0.4–0.9(–1.8) mm in diameter; soon sessile; proper margin black, conspicuously thickened and frequently fractured, always persistent, not excluded with age; disc black, epruinose, plane, not becoming convex with age; exciple very thick, distinctly differentiated into an inner and outer part, of the trachyspora-type (Fig. 41C,D), i.e. inner excipular hyphae large, leptodermatous, ± isodiametric cells, reddish brown, paraplectenchymatous (textura angularis), transient with the deep reddish brown hypothecium (leptoclinoides-brown, textura intricata), outer excipular hyphae shortcelled, cells globose, mesodermatous (textura globularis) and extremely carbonized with 338 large amounts of brown pigments (cf. elachista-brown), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown (cf. elachista-brown) pigment cap (cf. elachista-brown). Asci 8-spored, clavate, Bacidia-type. Ascospores (Fig. 41E–G) oblong to ellipsoid, sometimes constricted, with obtuse ends, not curved, (14.0–)18.8– [22.6]–26.5(–34.0) x (8.0–)8.7–[9.6]–10.5(–12.0) µm (n = 60); one-septate (occasionally additional septa forming at both ends of the spore cells), proper septum narrow, not thickening during spore ontogeny [Beltraminea (=Buellia)-type]; ornamentation coarsely rugulate to areolate; septum with septal pore canal, simple pore and undifferentiated pore plug; spore wall (Fig. 41G) differentiated into coarsely fractured, thick perispore (0.27– 0.97 µm), narrow intermediate layer (< 0.05 µm), thick proper spore wall (0.38–0.53 µm) and thick endospore (0.27–0.43 µm). Pycnidia rare to common, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform to fusiform, 10.0–13.0 x 0.5–1.0 µm (n = 20). Chemistry (Fig. 44).—Xanthones detected with HPLC: 4,5-dichloro-3-Omethylnorlichexanthone, 4,5-dichlorolichexanthone. All spot tests are negative (K–, P–, C–, KC–, CK–), but the thallus has a UV+ bright orange fluorescence. The thallus medulla is not amyloid, apothecia react amyloid in Lugol’s (always test with 339 concentrated Lugol's iodine or in the compound microscope; positive reactions can be very weak!). Substrate and ecology.—On sheltered siliceous rock (HCl–), usually close to rivers and streams. Distribution (Fig. 11).—This subtropical to tropical species barely reaches the southernmost Greater Sonoran Desert Region in southwestern Chihuahua (Barranca del Cobre, La Bufa, COLO – Weber L-65270). Additional collections at ASU were collected near Compostella in Nayarit, Mexico (outside the Greater Sonoran Desert Region). The species was first described from the Virgin Islands and included by Imshaug (1957) in his "Catalogue of the West Indian Lichens" (Cuba, Jamaica, Puerto Rico, Virgin Islands: St. Thomas, St. John). Notes.—Although this species has a unique apothecial anatomy, it has been confused with B. mamillana. Thalli of the two species are indeed very similar, but young apothecia of B. trachyspora emerge early during the ontogeny and are never remotely lecanorine (not even a thalline collar or a thalline veil has been observed in the material). Selected specimens examined.–MEXICO. NAYARIT. Nash 20751 (ASU). CHIHUAHUA. Weber L-65270 (COLO). 340 → FIGURE 37. Buellia concinna (A–F. light micrographs; G–I. TEM micrographs).—A. Granular thallus with sessile apothecia (Nash 22836).—B. Close-up of granules with apothecia (Nash 22836).—C. Cross section of the leptocline-type exciple, i.e. largely undifferentiated throughout and composed of intricately interwoven, mesodermatous hyphae (Nash 22836).—D. Mature ascospores with ± tapered ends and thickened septum (arrow; Nash 22836).—E. Premature, ellipsoid pale ascospore with distinct septum thickening, and mature, crescent-shaped ascospore with deeply pigmented wall and moderately thickened septum (Nash 11791).—F. Overmature, ± citriform ascospore (Nash 22836).—G. Premature ascospore (Nash 14655): (as) ascus wall; (s) mucilaginous sheath; (1) perispore; (2) intermediate layer; (3) proper spore wall; (4) endospore (at this stage of the ontogeny the proper spore wall and the endospore are not yet fully differentiated).—H. Mature ascospore (Nash 14655).—I. Wall layers in a mature ascospore (Nash 14655): (as) ascus wall; (s) mucilaginous sheath; (1) perispore; (2) intermediate layer; (3) proper spore wall; (4) endospore (at this stage of the ontogeny the proper spore wall and the endospore are fully differentiated). 341 342 → FIGURE 38. Buellia halonia (Nash 41434; A–F. light micrographs; G–L. TEM micrographs).—A. Areolate thallus with immersed sessile apothecia.—B. Close-up of areoles and apothecia: immature apothecia are immersed and large chunks of thalline material (arrows) often remains attached during emergence.—C. Cross section of the leptocline-type exciple.—D. Detail of the outer exciple: mesodermatous hyphae are not considerably swollen at the apices and ± interwoven throughout the exciple.—E. Ascus with eight mature ascospores (arrow indicates the septum thickening).—F. Mature ascospore with distinct septum thickening (arrow).—G. Immature ascospore.—H. Wall layers in an immature ascospore: (as) ascus wall; (1) outer layer (perispore and mucilaginous sheath not distinctly differentiated); (2) inner layer (proper wall and endospore not distinctly differentiated).—I. Mature ascospore: the septum thickening of the endospore is not yet developed.—J. Pycnidium.—K. Conidiophores with bacilliform conidia, predominantly apically formed.—L. Wall layers in a mature ascospore: (as) ascus wall; (s) mucilaginous sheath; (1) perispore; (2) intermediate layer; (3) proper spore wall; (4) endospore. 343 344 → FIGURE 39. Buellia mamillana (A–E. light micrographs, F–G. TEM micrographs).— A. Rimose thallus with immersed to ± sessile apothecia (Nash 28439).—B. Close-up thallus and apothecia (Nash 25875; arrows indicate the thalline part of the margin in some premature apothecia).—C. Cross section of an immature apothecium (Nash 25604) with a distinct thalline outer exciple [c. 60 µm wide; (c) cortex; (a) algae], and a narrow (c. 10 µm wide) inner exciple (arrows).—D. Cross section of a premature apothecium (Nash 25604): the proper exciple (arrows) is expanding (c. 30 µm wide) while the outer, thalline exciple (th) becomes excluded (c. 35 µm wide).—E. Mature ascospores with rugulate ornamentation (Nash 25604).—F. Mature ascospore (Nash 37836).—G. Wall layers in a mature ascospore (Nash 37836): (as) ascus wall; (s) mucilaginous sheath; (1) perispore; (2) intermediate layer; (3) proper spore wall; (4) endospore. 345 346 → FIGURE 40. Buellia subaethalea [A–D. light micrographs (Arsène 3654–lectotype), E– I. TEM micrographs (Ryan 27029)].—A. verrucose-areolate thallus.—B. Close-up of the verrucose-areolate thallus with ± immersed to sessile apothecia.—C. Cross section of a mature apothecium with aethalea-type exciple.—D. Mature, rugulate ascospore with two distinctly darkened bands (arrows).—E. Ascus with immature ascospores.—F. Immature ascospore: (as) ascus wall; (1) outer layer (perispore and mucilaginous sheath not distinctly differentiated); (2) inner layer (proper wall and endospore not distinctly differentiated).—G. Wall in an immature ascospore.—H. Wall layers in a mature ascospore: (s) mucilaginous sheath; (1) perispore; (2) intermediate layer; (3) proper spore wall; (4) endospore.—I. Mature ascospore. Note: the two bands, which are distinctly visible in the light microscope (Fig. 4A) do not correspond with any distinct wall structure in the TEM (Fig 4F–I). 347 348 → FIGURE 41. Buellia trachyspora (Nash 20750; A–E. light micrographs, F–G. TEM micrographs).—A. Rimose thallus.—B. Close-up of apothecia.—C.Cross section of a mature apothecium with trachyspora-type exciple: (1) outer, strongly carbonized exciple; (2) inner, paraplectenchymatous exciple; (3) hypothecium extending into the inner exciple.—D. Detail of the outer and inner exciple: (1) strongly carbonized cells of the outer exciple cannot be distinguished from one another; (2) leptodermatous, ± isodiametric cells of the inner exciple.—E. Mature ascospores with a strongly rugulate to areolate ornamentation.—F. Wall layers in a mature ascospore: (s) mucilaginous sheath; (1) perispore; (2) intermediate layer; (3) proper spore wall; (4) endospore. 349 350 → FIGURE 42. HPLC chromatograms of Buellia concinna (Nash 38098) and Buellia halonia (Nash 17166). 351 352 → FIGURE 43. HPLC chromatograms of Buellia mamillana (Nash 29793) and Buellia subaethalea (Nash 25353). 353 354 → FIGURE 44. HPLC chromatogram of Buellia trachyspora (Weber L-65279). 355 356 KEY TO THE SPECIES 1 Inner exciple distinctly paraplectenchymatous, with large, leptodermatous isodiametric cells; outer exciple very strongly carbonized (small globular cells barely distinguished even in thin microtome sections); spores with a coarsely rugulate to areolate ornamentation (clearly visible at 400x) .................................... ................................................................................................. Buellia trachyspora Inner and outer exciple not as above; spores smooth to rugulate but not coarsely areolate (ornamentation, if present, barely visible at 400x) .................................. 2 2(1) Thallus granular to bullate or verrucose-areolate, i.e. entirely composed of small granules or areoles with a distinctly granular surface ........................................... 3 Thallus rimose to areolate, not verrucose or granular ........................................... 4 3(2) Mature spores oblong to ellipsoid, with obtuse ends, not curved, with two spore cells covered by a broad, darkened band; thallus granular-areolate to verrucose; ... ................................................................................................. Buellia subaethalea Mature spores ellipsoid to ± citriform, often with ±tapered ends, frequently curved (crescent-shaped), the two spore cells never covered by a darkened band; thallus granular to minutely bullate; ....................................................... Buellia concinna 4(2) Apothecial disc deep brown, rarely blackened; young apothecia remaining immersed or, if emergent, distinctly lecanorine, i.e. with a distinct thalline margin; thalline margin darkening and eventually excluded; apothecia becoming lecideine with age; ascospores rugulate ................................................. Buellia mamillana 357 Apothecial disc black, not brownish black; young apothecia sometimes emerging with a thalline veil but never distinctly lecanorine; ascospores smooth to inconspicuously microrugulate ............................................................................. 5 5(4) Thallus thick, areolate; usually with a distinct black prothallus along the thallus margin; apothecia pruinose or epruinose; exciple thick, of intricately intertwined hyphae, carbonized by a diffuse, aeruginose (HNO3+ violet) pigment; conidia bacilliform to fusiform .................................................................. Buellia halonia Thallus thin, rimose to rimose-areolate; with or without a faint black prothallus; exciple thin, of narrow, parallel hyphae with swollen end cells, carbonized by a brown, non-aeruginose pigment (HNO3–); conidia filiform .................................... ..................................................................................................... Buellia prospersa DISCUSSION Thallus.—Even though all species can be characterized by xanthones, thallus color varies considerably among the species. Some species like B. prospersa and B. mamillana may have very pale thalli and the presence of xanthones in these thalli may thus be overlooked. Thalli of B. subaethalea and B. concinna become sordid yellow with prolonged herbarium storage. Buellia halonia is usually greenish yellow rather than pale yellow. The presence of xanthones in the thalli is generally indicated by a yellow to orange fluorescence in longwave (c. 350 nm) ultraviolet light. Intensity of this fluorescence, however, varies from pale to very bright even among thalli of the same species. Buellia concinna and B. halonia both have a C+ and KC+ orange thallus but the 358 other species usually have no indicative C (or KC) reaction event though their thalli contain xanthones. Thallus morphology for each of the species is quite distinct and most species can usually be recognized even without anatomical analysis. Buellia prospersa has thin, usually inconspicuous, rimose thalli (see Bungartz et al. 2004b). In comparison thalli of B. halonia are much thicker and distinctly areolate (Fig. 38A,B). The thallus of Buellia concinna is composed of small and ± dispersed granules (Fig. 37A,B), whereas B. subaethalea has a verrucose surface of an essentially areolate thallus (Fig. 40A,B). Buellia mamillana and B. trachyspora have very similar smooth, rimose thalli which are finely fissured (Figs. 39A,B and 41A, B). These two species are difficult to distinguish by their thalline characters alone and have thus been confused. Apothecia.—Buellia trachyspora and B. mamillana have apothecia which are unique among species in the genus Buellia. Apothecia of B. trachyspora become very large (up to 1.8 mm in diameter). Unlike in B. mamillana these large apothecia very soon emerge and become sessile. They remain distinctly flattened, with a plane disc and a thick margin (Fig. 41B). Due to the extreme carbonization of the outer exciple, this margin often becomes coarsely fractured in dry herbarium specimens. The carbonized cells of the outer exciple are small and ± globose and can barely be distinguished because of their strong carbonization. The inner exciple is entirely composed of large, isodiametric cells and thus distinctly paraplectenchymatous (Fig. 41C,D). Because of the angular shape of these cells the plectenchyma may be referred to as textura angularis. The inner exciple of B. trachyspora is entirely composed of thin-walled cells which are not carbonized. In 359 contrast, many other Buellia species have an outer exciple of ± angular cells, which are smaller, carbonized and mesodermatous. This texture has also been referred to as textura angularis but is quite different from the texture seen in B. trachyspora (Scheidegger 1993; Bungartz & Nash 2004a; 2004b; Bungartz et al. 2004b). The exciple of B. trachyspora is therefore unique and cannot be assigned to any of the types described by Scheidegger (1993). Buellia mamillana is the only species currently known from the Sonoran Region with apothecia developing a distinct thalline margin (Fig. 39B,C,D). Other species currently included in Buellia s.l. may have some irregular thalline collar (Bungartz & Nash III 2004a) or a thalline veil (Bungartz & Nash III 2004c), but only in Buellia mamillana do the young apothecia develop a distinct lecanorine margin with a cortex and algal layer embracing the young apothecium (Fig. 39C). As a result, the species was originally described as a Rinodina. It is unusual that this initially well developed thalline margin subsequently becomes excluded and replaced by hyphae from the proper exciple (Fig. 39D). During this process the deep brown pigmentation of the disc extends more and more across the initially pale margin. This shift in pigmentation is a result of internal growth processes. The inner exciple expands and the thalline outer part becomes excluded. During this ontogeny, apothecia may even have a lecanorine exciple on one side, and a lecideine exciple on the other side of a cross section. This ontogeny is very unusual. Weber (1986), who transferred the species into Buellia, did not describe this unusual ontogeny. He interpreted the apothecia as “clearly lecideine”, possibly because older apothecia indeed become lecideine. 360 Apothecia of the other species are not as unusual. Scheidegger (1993) already described the intricately interwoven excipular hyphae of B. halonia (Fig. 38C,D) and assigned it to the leptocline-type. This exciple type is not significantly differentiated throughout. It is initially less strongly pigmented inside but soon becomes evenly pigmented. Buellia concinna has an exciple very similar to B. halonia, but lacks the aeruginose pigment. Like in B. halonia excipular hyphae of B. concinna are ± evenly pigmented, little differentiated and intricately interwoven (Fig. 37C). Despite this, Scheidegger (1993) assigned the exciple of B. concinna to the dispersa-type, which he described as composed of hyphae radiating from a deep reddish brown hypothecium, with a ±hyaline transition zone and a strongly carbonized outer layer of moderately swollen cells. The exciple of B. concinna does not show this differentiation and thus should be assigned to the leptocline-, not the dispersa-type. The exciple of B. subaethalea is less prominent and the inner hyphae are thinner than in B. halonia or B. concinna. In B. subaethalea only the outer cells are usually ± swollen and strongly pigmented (Fig. 40C). The exciple of B. prospersa is very similar (Bungartz et al. 2004b). Both species belong structurally to the aethalea-type, even though they lack the aeruginose pigment cinereorufa-green. Asci and Ascospores.—All species have Bacidia-type asci and a dark hypothecium, characters typically associated with the Buellia-clade rather than the Rinodina-clade (Helms et al. 2003). Ascospores of B. trachyspora have no distinct septum thickening and may thus be assigned to the Buellia (=Beltraminea)-type, even though the spores are 361 usually ± constricted along the septum and younger spores show some inconspicuous swelling where the septum forms. The ascospores of all other species have a distinct septum thickening at least during some part of their ontogeny. They can thus be referred to the Physconia-type. In most species this septum thickening is ephemeral, i.e. it is most pronounced in premature spores but becomes reduced in mature and overmature spores. Only B. prospersa has a septum, which usually is more persistent and considerably thicker than in any of the other species. These spores could, therefore, also be referred to as Orcularia-type. None of these spore types refer to strictly distinctive categories but represent an idealized concept of an inherently dynamic and transient development of the spore. For example, even though both B. concinna and B. subaethalea are formally assigned to the same spore type, their ontogenies are distinctly different from all the other species. Spores of B. concinna develop a very distinctive shape (Fig. 37D,E,F). The premature to mature ascospores are characteristically curved and have ± tapered or pointed ends. These spores are best described as a “half-moon” or crescent-shaped (Fig. 37E). The characteristic shape is not always observed because some spores do not become curved. These spores nevertheless develop ± tapered ends and thus appear ± citriform. All other species have spores with obtuse ends. Old spores of B. subaethalea frequently become ± constricted but are more importantly characterized by two, broad and distinctly darkened bands across each of the two spore cells (Fig. 40D). In the light microscope these bands feign lateral wall thickenings, which must not be confused with distinct structural lateral thickenings in 362 Callispora-type ascospores. Ultrastructural observations currently suggest that the “banding” does not correspond with structural layers of the lateral cell walls. However, comparing the ultrastructure of ascospores at different ontogenetical stages may elucidate whether these patterns correspond to some ultrastructural differentiation of the spore wall. Similar “bands” are known from ascospores of Rinodina and may be caused by a similar phenomenon to the torus. Spore ornamentation is a result of the perispore differentiation. Thus, the ultrastructure of this outer spore wall layer corresponds to the ornamentation observed in the microscope. In B. trachyspora the perispore appears coarsely fractured in the TEM (Fig. 41F,G) and a rugulate to areolate ornamentation (Fig. 41E) is visible in light microscope even at 400 x magnification. In other species, spore ornamentation can also be observed but is much less distinct. Buellia subaethalea also has rugulate ascospores (Fig. 40D,I), but the ornamentation is barely visible at 400 x and distinct only at 1000 x magnification. The rugulate ornamentation of Buellia mamillana is even less distinct (Fig. 39E) and the perispore is not strongly fractured (Fig. 39G). Finally, B. halonia and B. prospersa both have a microrugulate ornamentation. This fine ornamentation is quite distinct at 1000x in B. prospersa but barely visible in B. halonia. Conidiomata.— The conidiomata of all species are quite similar. They develop in urceolate to globose pycnidia (Fig. 38J), which are almost entirely occupied by conidiophores. These conidiophores are branched and predominantly form conidia at the apices (Fig. 38K). Some conidia are also formed intercalary on short bayonet-like protrusions. The conidia do, however, not anastomose at their apices and thus resemble 363 more closely type V instead of type VI (Vobis 1980). Buellia concinna has the shortest, ± ellipsoid conidia. Buellia halonia and B. subaethalea have slightly longer, bacilliform conidia. Conidia of B. mamillana and B. trachyspora are bacilliform to fusiform and B. prospersa has filiform conidia. Because of its long conidia, B. prospersa has also been accommodated in the genus Amandinea [as Amandinea lecideina (H. Mayrhofer & Poelt) Scheid. & H. Mayrhofer]. The segregation of an entire genus on this single character is, however, problematic (Bungartz et al. 2004b). Chemistry (Figs. 42–43).—Detailed information on the thallus chemistry of the species is provided in the species descriptions. Buellia concinna and B. halonia are the only species which contain asemone, thiophanic acid, and arthothelin together with the closely related isoarthothelin. The other species contain a variety of different lichexanthones, which are also present in B. concinna and B. halonia. Buellia subaethalea is the only species which contains a single xanthone (4,5dichchlorolichexanthone). In contrast, B. mamillana has the most complex chemistry, regularly with a variety of xanthones, depsides and depsidones. This chemical variation does not appear to correlate with any morphological differences. Distribution.—Of all the species examined only B. halonia shows a distinctly coastal, maritime distribution. Buellia concinna and B. subaethalea are montane to subalpine species and possibly occur in the same habitats. Buellia mamillana and B. trachyspora show subtropical affinities. Buellia mamillana extends far into the Sonoran Desert Region and typically grows along mountain streams. Buellia trachyspora grows in similar habitats but barely extends into the southernmost part of the Sonoran Region. 364 ACKNOWLEDGMENTS We are grateful to Dr. John Sheard, University of Saskatchewan, Canada and Scott Bates, for reviewing the manuscript. James Lendemer, Academy of Natural Sciences, Philadelphia also provided some valuable advice for the manuscript. Loans from the following herbaria to ASU are greatly appreciated: BM, C, CANL, COLO, FH, H, hb. Scheidegger, hb. Kalb, hb. Sheard, L, M, MICH, MSC, SBBG, TUR, UPS, W. Dr. Ulrik Søchting and Dr. Eric Hansen, University of Copenhagen (Denmark) and Dr. Seppo Huhtinen, University of Turku (Finland) provided valuable advice on the correct typification of Buellia trachyspora. Several lichenologists quickly responded to a literature search for rare taxonomic literature posted on the lichen list server. The study was supported by a Research Grant in Plant Systematics from the International Association of Plant Taxonomists (IAPT) and National Science Foundation Awards to ASU (DEB-0103738, DEB-9701111). A research visit of the first author to MSU was supported by a National Science Foundation Award (DBI-0237401). LITERATURE CITED BOULY DE LESDAIN, M. 1914. Lichens du Mexique (états de Puebla et du Michoacan) recueillis par le frère Arsène Brouard. imp. I. Escalante, S.A. BUNGARTZ, F. & T. H. NASH III. 2004a. Buellia subalbula (Nyl.) Müll. Arg. and B. amabilis de Lesd., two species from North America with one-septate ascospores: A comparison with Buellia ["Diplotomma"] venusta (Körb.) Lettau. pp. 49-66. In RAMBOLD, G. & P. DÖBBELER (eds.), Lichenological Contributions. Festschrift in 365 honor of Hannes Hertel on occasion of his 65th birthday, Bibliotheca Lichenologica, 88. Gebrüder Borntraeger, Stuttgart. ———. 2004b. Buellia turgescens is synonymous with Buellia badia and must not be included in Amandinea. The Bryologist 107: 21-27. ———. 2004c. The Buellia aethalea-Group in the Greater Sonoran Desert Region with some reference to similar species known from North America. The Bryologist, submitted for publication February 2004. BUNGARTZ, F., C. SCHEIDEGGER & T. H. NASH III. 2002. Buellia dispersa A. Massal., a variable lichen species from semi-arid to arid environments of North America and Europe. pp. 19-35. In LLIMONA, X., H. T. LUMBSCH & S. OTT (eds.), Progress and Problems in Lichenology at the Turn of the Millennium, Bibliotheca Lichenologica, 82. J. Cramer, Berlin, Stuttgart. BUNGARTZ, F., T. H. NASH III & B. D. RYAN. 2004. Morphology and anatomy of chasmolithic versus epilithic growth: a taxonomic revision of inconspicuous saxicolous Buellia species generally ascribed to the "Buellia punctata"-group from the Sonoran Desert Region. Canadian Journal of Botany, in press. CULBERSON, C. F. & H. KRISTINSSON. 1970. A standardized method for the identification of lichen products. Journal of Chromatography 46: 85-93. CULBERSON, C. F. & A. JOHNSON. 1982. Substitution of methyl tert.-butyl ether for diethyl ether in standardized thin-layer chromatographic method for lichen products. Journal of Chromatography 238: 438-487. EGAN, R. S. 2001. Long-term storage of TLC data. Evansia 18: 19-20. 366 ELIX, J. A., M. GIRALT & J. H. WARDLAW. 2003. New chloro-depside from the lichen Dimelaena radiata. Bibliotheca Lichenologica 86: 1-7. FOUCARD, T., R. MOBERG & A. NORDIN. 2002. Buellia, pp. 11-25, (with an appendix on pp. 70-71). In AHTI, T., P. M. JØRGENSEN, H. KRISTINSSON, R. MOBERG, U. SØCHTING & G. THOR (eds.), Physciaceae. TH-tryck AB, Uddevalla. HELMS, G., T. FRIEDL & G. RAMBOLD 2003. Phylogenetic relationships of the Physciaceae inferred from rDNA sequence data and selected phenotypic characters. Mycologia 95: 1078-1099. IMSHAUG, H. A. 1955. The lichen genus Buellia in the West Indies. Farlowia 4: 473-512. ———. 1957. Catalogue of West Indian Lichens. Bulletin of the Institute of Jamaica, Science Series 6: 1-153. MALME, G. O. A. 1902. Die Flechten der ersten Regnellschen Expedition. II. Die Gattung Rinodina (Ach.) Stiz. Bihang till Kongl. Svenska Vetenskaps-Akademiens Handlingar 28: Jan-53. MEYER, B. & C. PRINTZEN. 2000. Proposal for a standardized nomenclature and characterization of insoluble lichen pigments. Lichenologist 32: 571-583. MIETZSCH, E., H. T. LUMBSCH & J. A. ELIX. 1994. WINTABOLITES (Mactabolites for Windows). Users manual and computer program. University Essen, Essen. NORDIN, A. 2000. Taxonomy and phylogeny of Buellia species with pluriseptate spores (Lecanorales, Ascomycotina). Symbolae Botanicae Upsalienses 33: 1-117. ORANGE, A., P. W. JAMES & F. J. WHITE. 2001. Microchemical Methods for the Identification of Lichens. British Lichen Society, London. 367 SCHEIDEGGER, C. 1993. A revision of European saxicolous species of the genus Buellia De Not. and formerly included genera. Lichenologist 25: 315-364. SCHEIDEGGER, C. & B. RUEF. 1988. Die xanthonhaltigen, gesteinsbewohnenden Sippen der Flechtengattung Buellia De Not. (Physciaceae, Lecanorales) in Europa. Nova Hedwigia 47: 433-468. SHREVE, F. & I. L. WIGGINS. 1964. Vegetation and Flora of the Sonoran Desert. Stanford University Press, Stanford. SINGH, S. R. & D. D. AWASTHI. 1981. The lichen genus Buellia in India. Biological Memoirs 6: 169-196. TUCKERMAN, E. 1888. A synopsis of the North American lichens. Part. II. Comprising the Lecideacei, and (in part) the Graphidacei., New Bedford, Massachusetts. VAINIO, E. A. 1915. Additamenta ad Lichenographiam Antillarum illustrandam. Ann. Sci. Fenn. ser A. 6: 1-226. VOBIS, G. 1980. Bau und Entwicklung der Flechten-Pycnidien und ihrer Conidien. Bibliotheca Lichenologica 14: 1-141. WEBER, W. A. 1986. The lichen flora of the Galapagos Islands, Ecuador. Mycotaxon 27: 451-497. WHITE, F. J. & P. W. JAMES. 1985. New guide to microchemical techniques for the identification of lichen substances. British Lichen Society Bulletin 57: 1-41. ZAHLBRUCKNER, A. 1931. Catalogus Lichenum Universalis. Borntraeger, Leipzig. 368 PUBLICATION 8 (Mycotaxon, submitted March 2004): New and previously not recorded saxicolous species of Buellia s.l. with one-septate ascospores from the Greater Sonoran Desert Region FRANK BUNGARTZ Arizona State University, School of Life Sciences, P.O. Box 87 4601, Tempe, AZ 85287–4601, U.S.A. frank.bungartz@asu.edu. Abstract. The following species of Buellia s.l. from the Greater Sonoran Desert Region are described new to science: Buellia nashii, B. navajoensis, B. regineae and B. sheardii. A valid description is provided for B. lepidastroidea, a name introduced by Imshaug, but never validly published. Buellia argillicola is reported from the Sonoran Region for the first time. A single record of the rare and unusual B. vilis is reported from Arizona. Buellia novomexicana and B. fusca are synonymized with B. tyrolensis. Detailed descriptions are provided for all these species. In addition a dichotomous key includes all saxicolous species with one-septate ascospores currently known from the Greater Sonoran Desert Region. Zusammenfassung. Die folgenden neuen Arten der Sammelgattung Buellia s.l. werden aus der Sonoraregion beschrieben: Buellia nashii, B. navajoensis, B. regineae and B. sheardii. Eine weitere Art ist B. lepidastroidea , die bereits von Imshaug vorgeschlagen, aber nicht gültig publiziert wurde, wird hier mit einer gültigen 369 Artdiagnose versehen. Buellia argillicola wird erstmals aus der Sonoraregion nachgewiesen. Ein Einzelfund der seltenen und ungewöhnlichen Art B. vilis wird aus Arizona gemeldet. Buellia novomexicana und B. fusca werden mit B. tyrolensis synonymisiert. Alle diese Arten sind hier detailliert beschrieben. Außerdem ist ein dichotomer Schlüssel der Publikation beigefügt, welcher die Bestimmung aller bisher aus der Region bekannten, saxikolen Arten mit zweizelligen Sporen ermöglicht. INTRODUCTION A revision of the saxicolous species of Buellia s.l. with one-septate ascospores from the Greater Sonoran Desert Region currently accepts a total of thirty-one species. Several of these species have never been reported from the area or are not yet described. Descriptions of these new species and records are given here. A complete treatment of the genus will be included in the third volume of the Sonoran Desert Lichen Flora (Nash III et al. 2002) but a key to all species is published here to allow identification of all species currently known from this region. The species described here are not necessarily members of what might be considered the core group of Buellia De Not. (Bungartz & Nash III 2004c). Traditionally specimens, which are related to Buellia aethalea (Ach.) Th. Fr. have been associated with Buellia s.str. but the current Botanical Code (Greuter et al. 2000) lists Buellia disciformis (Fr.) Mudd, a species with distinct characters of Hafellia Kalb, H. Mayrhofer & Scheid., as the type. There is currently some discussion to change this typification, but a proposal by Moberg et al. (1999) is not presently accepted. Until the outcome of this proposal is decided, a broad concept of Buellia s.l. is therefore adopted. Taxonomic affinities of the 370 species described, are discussed in more detail within brief notes for each species. The genus concept of Buellia is currently in a state of much disarray (Nordin 2000; Eriksson et al. 2002a) and the question, which genera will ultimately be recognized should best be addressed using molecular tools to evaluate classical data. METHODS All specimens were examined with light microscopy using hand- and cryosections. Both conventional bright field microscopy (BF) as well as differential interference contrast (DIC) were used. Selected specimens were also studied with transmission electron microscopy (TEM) according to a protocol described in detail by Bungartz et al. (2002). To improve dehydration and infiltration this protocol has been modified according to Bungartz & Nash (2004a). All specimens were spot tested and routinely examined with standardized thin-layer chromatography (Culberson & Kristinsson 1970; Culberson & Johnson 1982; White & James 1985; Orange et al. 2001). TLC-Plates were interpreted with the computer program WINTABOLITES (Mietzsch et al. 1994), and scanned for permanent record Egan (2001). In addition a subset of specimens was analyzed by Prof. J. A. Elix from the Australian National University in Canberra using standardized High Performance Liquid Chromatography (HPLC, Feige et al. 1993). Spores measurements are given according to Nordin (2000). Pigment names follow Meyer & Printzen (2000). Detailed specimen information about all collections deposited at ASU is available at: http//seinet.asu.edu/collections/selection.jsp. 371 TAXONOMIC DESCRIPTIONS BUELLIA ARGILLICOLA de Lesd., Ann. Cryptog. Exot. II: 243. 1929. TYPE: MEXICO. MÉXICO. San Jerónimo. On volcanic rocks (pebbles embedded in soil) [original label data: Estado de Distrito Federal. San Jerónimo, Sobre Piedra], 19°13'30''N, 100°01'30''W, 2598 m alt., Pedro (Pierre) Lyonett no. 110 (MEXU! – neotype selected here). Taxonomic note.—No type material from the original collections was found. It can be assumed that de Lesdain’s holotype collection, originally located in Dunkerque (France), has been destroyed during the Second World War. The specimen selected as a neotype was collected at the type locality and agrees well with the description in the protologue. Thallus (Fig. 45A,B).—crustose, thin, usually growing in distinct circular patches, subeffigurate, several thalli often confluent, epilithic; continuous to rimose with fine fissures; prothallus distinct, delimiting the thallus as a black outline; thallus surface matt and chalky, dull, usually white, rarely gray, heavily pruinose, phenocorticate; entire thallus filled with an abundance of calcium oxalate (H2SO4+ forming clusters of needle shaped crystals). Apothecia lecideine, (0.3–)0.5–0.8(–1.1) mm in diameter, remaining immersed to indistinctly adnate; proper margin thick, persistent, not excluded with age, black, usually covered with a dense, fine, white pruina; disc black, plane, not becoming convex with age, usually covered with a dense, fine, white pruina; proper exciple similar to the aethalea-type (Fig. 45C) sensu Scheidegger (1993), i.e. inner exciple almost entirely reduced, hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), similar in structure and orientation to the paraphyses, transient with the deep reddish brown 372 hypothecium (leptoclinoides-brown, textura intricata), outer exciple expanded, globular cells strongly swollen (± textura oblita) and moderately carbonized with a brown pigment (cf. elachista-brown), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown (cf. elachista-brown) pigment cap. Asci 8-spored, clavate, Bacidiatype. Ascospores (Fig. 45D,E) — oblong to ellipsoid, usually not constricted, with obtuse ends, not curved, (12.0–)15.0–[16.8]–18.5(–20.0) x (6.0–)6.5–[7.3]–8.1(–9.0) µm (n = 60), one-septate, proper septum narrow, not thickening during spore ontogeny, lateral wall thickenings absent [Beltraminea (=Buellia)-type]; ornamentation rugulate. Pycnidia rare, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicariatype (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 4.0–6.5 x 1.0–1.5 µm (20). Chemistry.—With the depsidones norstictic, connorstictic, stictic and hypostictic acid and one unknown substance (RF 7, not visible in natural light, UV+ orange). Thallus and medulla K+ faint yellow (yellow solution, but not orange, needle-shaped crystals observed in the microscope), P+ faint yellow, C–, KC–, CK–. UV+ pale white. Thallus and apothecia react amyloid with Lugol’s (always test with concentrated Lugol's iodine or in the compound microscope; positive reactions can be very weak!). 373 Substrate and ecology.—The type specimen grows on pebbles embedded in calcareous (HCl+) soil (= tépetate), the only known Sonoran specimens grow on limestone (strongly HCl+). Distribution (Fig. 7).—Known only from very few localities. Notes.— Specimens of B. argillicola are superficially similar to B. subalbula (Nyl.) Müll. Arg. or B. venusta (Körb.) Lettau. The thalli of B. argillicola are, however, more continuous and less extensively fissured. Apothecia of B. argillicola have a plane disc with a persistent lecideine margin and no thalline collar or veil. Internally the structure resembles the aethalea-type but the outer exciple is much more prominent and persistent. Both B. subalbula and B. venusta have a thin, largely reduced exciple, which usually becomes excluded by the swelling of the disc. In B. venusta a thalline collar is common and it can sometimes be present in B. subalbula. Buellia subalbula is restricted to the coast whereas B. argillicola is known from inland localities. Buellia subalbula is characterized by a fuscous brown to deeply aeruginose exciple, which always reacts HNO3+ violet. Buellia venusta and B. argillicola have no aeruginose exciple pigment. Buellia venusta has pluriseptate, B. argillicola one-septate ascospores. Representative specimens examined.—MEXICO. SINALOA. Nash 12152 (ASU). BUELLIA LEPIDASTROIDEA Imshaug ex Bungartz sp. nov. Thallus saxicolus, crustaceus, areolatus vel bullato-areolatus, crassus, eburneus, cum hypothallo atero. Apothecia sessilia, lecideina, marginibus propriis exclusis. Excipulum tenuis, fulvum, sine pigmento aeruginoso, carbonaceum. Hymenium interspersum guttae 374 olei. Asci 8-spori. Sporae uniseptae, oblongae vel ellipsoideae, septibus incrassatus, 7–17 x 5-8 µm. Pycnidia urceolata vel globosa. Conidia bacilliformes, 2-7 x 1-2 µm. Thallus atranorina, ± diploicina et ± fulgidina continens. Medulla non- amyloideus. TYPE: MEXICO. BAJA CALIFORNIA. Isla Cedros, ridge crest overlooking western shore and adjacent canyon to the E at the NW corner of the island, 28º22'00" N, 115º15'30"W, ca. 300 m alt., on acidic rock, 19 March 1994, Nash 34458 (ASU–holotype designated here!). Taxonomic note.—The species name was first used by Imshaug (1951) in his dissertation but was never validly published. Thallus (Fig. 46A,B) crustose, thin to moderately thickened, ± continuous or becoming dispersed, epilithic; areolate to subsquamulose or bullate; prothallus delimiting the thallus margin, distinctly blackened to pale gray, rarely white and indistinct; thallus surface matt and dull, not shiny, usually ivory, rarely beige, epruinose or rarely with a fine pruina, phenocorticate; without Ca-oxalate crystals in the medulla (H2SO4–). Apothecia lecideine, (0.3–)0.5–0.8(–1.1) mm in diameter, soon sessile; proper margin thin, black, rarely persistent, usually excluded with age; disc black, epruinose, plane, soon becoming convex with age; proper exciple of the dispersa-type (Fig. 46C) sensu Scheidegger (1993), i.e. inner excipular hyphae distinct, not reduced, pigmented, prosoplectenchymatous (textura oblita), extending from the deep reddish brown hypothecium (leptoclinoides-brown, textura intricata); outer excipular hyphae shortcelled, cells angular, distinctly swollen (textura angularis) and usually ± carbonized with 375 various amounts of brown pigments (cf. elachista-brown), pigmentation continuous with the epihymenium; hymenium hyaline, inspersed with oil droplets; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachistabrown). Asci 8-spored, clavate, Bacidia-type (Fig. 46F). Ascospores (Fig. 46D,G,H) oblong to ellipsoid, usually not constricted, with obtuse ends, not curved, (7.0–)11.4– [13.2]–15.0(–17.0) x (5.0–)6.0–[6.7]–7.4(–8.0) µm (n = 60); one-septate, proper septum briefly thickened but soon becoming reduced during spore ontogeny, lateral wall thickenings absent (± Physconia-type); ornamentation absent (not visible in DIC); septum with septal pore canal, simple pore and undifferentiated pore plug; spore wall (Fig. 46G,H) differentiated into smooth, thin perispore (0.06–0.07 µm), narrow intermediate layer (< 0.03 µm), thick proper spore wall (0.34–0.54 µm) and moderately thickened endospore (0.12–0.21 µm). Pycnidia rare, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 2.0–7.0 x 1.0–2.0 µm (41). Chemistry.—With the depsides atranorin and chloroatranorin and the depsidones diploicin, 3-dechlorodiploicin, fulgidin and isofulgidin. HPLC–artefacts: haematommic acid, methyl β-orsellinate (HPLC by J. Elix, Canberra). Thallus and medulla K+ yellow, P+ orange, C–, KC–, CK–. UV+ pale ivory. The thallus medulla and cortex does not react amyloid, apothecia are amyloid in Lugol’s (always test with concentrated Lugol's iodine or in the compound microscope; positive reactions can be very weak!). 376 Substrate and ecology.— On a variety of siliceous (HCl–) rock substrates. Distribution (Fig. 8).— Common along the coast of southern California and Baja California. Notes.—Thalli of B. lepidastroidea sometimes vaguely resemble B. dispersa or B. nashii. Buellia lepidastroidea is, however, strictly coastal and characterized by oil droplets in the hymenium. This inspersion is usually distinct, especially if thick apothecial sections are treated with 5% KOH. In thin sections, oil inspersion may be less conspicuous but nevertheless always present. Thalli growing close to the seashore and subject to spray often show more exuberant growth than sheltered thalli. Their thallus areoles become strongly inflated and apothecia thus appear “stalked”. The North American specimens fit well into the concept of Imshaug’s B. lepidastroidea and the species is described here to validate his name. The species belongs to a group of taxa with indistinct Physconia-type ascospores, a dispersa-type exciple and an oil-inspersed hymenium. This group of species is currently not well understood. Buellia lepidastroidea is very similar to European material of B. excelsa (Leight.) A.L. Smith. Scheidegger (1993) selected a lectotype for B. excelsa but the material could not be found at BM. Specimens identified as B. excelsa from Scheidegger’s private herbarium have a more distinctly brown thallus, contain only atranorin, not diploicin or fulgidin and were collected at high altitude, not close to the seashore. Scheidegger (1993, p. 349) mentions that the species is distributed “in dunes and in mountains”. In the Sonoran Desert Region B. lepidastroidea is clearly confined to coastal, maritime habitats and has not been found further inland or at high altitudes. Other taxa in the Buellia excelsa-group 377 are B. boergesenii Imshaug and B. jorgensis Zahlbr. (CHILE—M-0061335, Spindler 16158, det. Follmann). Buellia conspirans (Nyl.) Vain. is a saxicolous species similar to B. boergensii according to Imshaug (1955). It is currently listed in the North American Checklist (Esslinger 1997) as a synonym of B. curatellae Malme. Buellia curatellae is, however, a corticolous species with distinct lateral spore wall thickenings. It was transferred by Marbach (2000) into the genus Hafellia. Buellia lepidastroidea is treated here as a distinct species, but a more detailed revision of the Buellia excelsa-group is necessary to evaluate the taxa currently included in this group. Because of the oil inspersion in the hymenium, species of the Buellia excelsa– group show some affinities to the genus “Hafellia”. The presence of diploicin and related substances in B. lepidastroidea is unusual for Buellia s.str. but these substances have been reported from “Hafellia”. Scheidegger (1993) did, however, not include B. excelsa within “Hafellia” because the ascospores lack lateral wall thickenings and he did not find diploicin in B. excelsa. Buellia lepidastroidea usually can be easily distinguished from B. (“Hafellia”) regineae because of its thinner thallus, with smaller, more dispersed areoles, smaller apothecia and ascospores. Lateral wall thickenings are usually present in spores of B. regineae, but less distinct than in B. leptoclinoides (see notes on B. regineae). Thalli of B. lepidastroidea and B. excelsa are very similar to B. dispersa and B. nashii. All these species also share the same exciple anatomy and have indistinct Physconia-type ascospores. The angular thickening of the spore septum, which is characteristic for the Physconia-type ascospore, can only very briefly be observed in premature ascospores 378 (Fig. 46D) but becomes soon reduced in mature spores (Fig. 46G,H). Buellia dispersa and B. nashii, however, do not contain oil droplets within the hymenium. Representative specimens examined.––MEXICO. BAJA CALIFORNIA. Egan 13790 (OMA);. Kalb 24657 (hb. Kalb); Scheidegger Inv. Nr. 170-189, 71-95, 150-155 (hb. Scheidegger);. Weber L-43033, L-43014, L-43062, L-36575 (COLO); Wetmore 63852, 75765, 72431, 63498, 75873 (MIN); Nash 38242, 34370, 38241, 8688b, 34319, 4916, 34052, 38256, 8730, 25188, 38459, 34358, Ryan 2136, 21263 (ASU). BAJA CALIFORNIA SUR. Wetmore 72223 (MIN);. Marsh 6080, Nash 8947 (ASU). –– UNITED STATES. CALIFORNIA. Los Angeles Co. Bratt 9799, 10219, 11843, 11844 (SBBG); Hasse Exs. 155 (ASU); Weber L-42606, L-42823 (COLO). Santa Barbara Co. Bratt 7379, 7605 (SBBG). CALIFORNIA. Santa Barbara Co. Nash 32646, 33021, Ryan 31255b, 31270 (ASU). WASHINGTON. Kittitas Co. Ryan 16838 (ASU). BUELLIA NASHII Bungartz sp. nov. Similis Buelliae dispersae sed apotheciae cum pigmento aeruginoso et thallus acida norstictica et connorstictica vel acida stictica continens. Etymology.—The species is named in honor of my Ph.D. supervisor Dr. Thomas H. Nash III. TYPE: MEXICO. COAHUILA. 3 km W of the paved road, at the Delores sign, 26º00'00"N, 101º00'00" W (estimated coordinates), on volcanic rock, 20 March 1976, Nash 6742a (ASU! – holotype designated here; one isotypes at MEXU, one isotype at US). 379 Thallus (Figs. 47A,B) crustose, thick, ± continuous or becoming dispersed, epilithic; areolate to subsquamulose; prothallus absent; thallus surface matt and smooth to deeply fissured, dull or ± shiny, usually ivory, beige to deep brown or gray, rarely lead gray, with fine or coarse pruina, rarely epruinose, phenocorticate; with few, or rarely large amounts of calcium oxalate crystals (H2SO4+ forming clusters of needle shaped sulphate crystals). Apothecia lecideine, (0.4–)0.6–1.1(–1.2) mm in diameter, sessile; proper margin black, thin to thick, usually persistent, rarely excluded with age; disc black, usually epruinose, rarely with white pruina, plane, often becoming strongly convex with age; proper exciple of the dispersa-type (Figs. 47C) sensu Scheidegger (1993), i.e. inner exciple distinct, not reduced, hyphae pigmented, prosoplectenchymatous (textura oblita), extending from the deep reddish brown hypothecium (leptoclinoides-brown, textura intricata); outer excipular hyphae short-celled, cells angular, distinctly swollen (textura angularis) and carbonized with various amounts of a brown pigment (cf. elachista-brown); small amounts of a diffuse aeruginose pigment present but restricted to the outermost exciple cells (cinereorufa-green, HNO3+ violet); only the brown pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown). Asci 8-spored, clavate, Bacidia-type. Ascospores (Figs. 47D,E) blong to ellipsoid, rarely constricted with age, with obtuse ends, not curved, (11.0–)12.9– [14.5]–16.1(–19.0) x (6.0–)6.5–[7.3]–8.1(–9.0) µm (n = 60); one-septate, proper septum briefly thickened but very soon becoming reduced during spore ontogeny, lateral wall thickenings absent (± Physconia-type); ornamentation microrugulate. Pycnidiarare, 380 urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicariatype (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia (Fig. 47F) simple, bacilliform, 5.0–10.0x 1.5–2.0 µm (n = 20). Chemistry.—Typically with the depside atranorin and the depsidones norstictic (forming orange, needle-shaped crystals in the compound microscope) and connorstictic acid . Some specimens do not contain norstictic acid but instead are characterized by stictic and hypositictic acid. Specimens typically react K+ yellow to red (sometimes weak), P+ yellow (sometimes weak), C–, KC–, CK–. UV– (pale). The thallus is not amyloid, apothecia react amyloid in Lugol’s (always test with concentrated Lugol's iodine or in the compound microscope; positive reactions can be very weak!). Substrate and ecology.—On a variety of siliceous (HCl–) rock substrates, rarely also on sandstones with small amounts of carbonates (HCl+). Distribution (Fig. 8).—The species is widely distributed throughout the Sonoran Desert Region and adjacent areas, extending into NE Mexico, but absent from low desert elevations. Notes.—Thalli of B. nashii are very similar and almost as variable as those of B. dispersa (see the description of morphotype I in Bungartz et al. 2002). The two species nevertheless are reliably distinguished by their chemistry (2'-O-methylperlatolic acid in B. dispersa, vs. norstictic and/or stictic acid in B. nashii) and their different exciple pigmentation. No specimen with the chemistry of B. dispersa contains any traces of the 381 aeruginose, HNO3+ violet pigment cinereorufa-green. In specimens of B. nashii this pigment is always present but usually confined only to the outermost cells of the exciple. Specimens should always be tested with HNO3 for a reliable identification. Most specimens of B. nashii contain norstictic acid and are thus easily distinguished form B. dispersa because of the formation of distinct orange, needle-shaped crystals if KOH is applied to apothecial or thallus sections. Representative specimens examined.––MEXICO. BAJA CALIFORNIA. Nash 26285, 26320b (ASU). CHIHUAHUA. Nash 13817, 13766 (ASU). COAHUILA. Nash 6742c (ASU). UNITED STATES. ARIZONA. Cochise Co. Weber S8831 (COLO). Coconino Co. Boykin 2849, 2674, Nash 30677, 4980 (ASU). Gila Co. Nash 7386 (ASU). Graham Co. Nash 7661 (ASU). Maricopa Co. Nash 8411, 6266, 9850, 6659, 987, 5011 (ASU). Mohave Co. Nash 7303 (ASU). Pinal Co. Nash 6151 (ASU). Santa Cruz Co. Nash 39231 (ASU). Yavapai Co. Nash 34137, 34149, 34188, 6361, Ryan 26958 (ASU). CALIFORNIA. Inyo Co. Ryan 14881 (ASU). San Diego Co. Nash 7052 (ASU). COLORADO. San Juan Co. Nash 17757 (ASU). NEVADA. Lincoln Co. Ryan 15862 (ASU). NEW MEXICO. San Juan Co. Nash 16397, 16395, Rankert 117 (ASU). UTAH. Kane Co. Nash 5071, 5457, Nash 6610 (ASU). San Juan Co. Nebecker 2800 (ASU). Washington Co. Nash 15315 (ASU). BUELLIA NAVAJOENSIS Bungartz sp. nov. Thallus saxicolus, crustaceus, areolatus vel sublobatus, crassus, eburneus, sine hypothallo. Apothecia immersia vel adnatia, lecideina, marginibus propriis tenuis. Excipulum tenuis, fulvo-caeruleum, pigmentum aeruginosum continens, carbonaceum. 382 Asci 8-spori. Sporae uniseptae, oblongae vel anguste oblongae, septibus incrassatus 13–25 x 6-10 µm. Pycnidia urceolata vel globosa. Conidia bacilliformes, 3-4.5 x 1-1.5 µm. Thallus acida norstictica et connorstictica et substancia xanthonia continens. Medulla amyloideus. Etymology.—The species is named after the Native American Navajos and the type specimens were collected on Navajo Sandstone. TYPE: U.S.A. UTAH. Kane Co. About 16 km east of Kanab along U.S. Highway 89, in Johnson Canyon near the Kanab Stake Welfare Farm, ca. 1734 m, 37 02'14"N, 112 21'53"W, on Navajo Sandstone, Nebecker 1577 (ASU–holotype designated here!). Thallus (Fig. 48A,B) crustose, thick, ± continuous, epilithic; areolate to sublobate often forming rosettes; prothallus absent; thallus surface matt and dull, not shiny, usually ivory, beige, rarely with a pinkish tinge, usually strongly pruinose, rarely weak or absent, phenocorticate; entire thallus filled with an abundance of calcium oxalate (H2SO4+ forming needle shaped crystals). Apothecia lecideine, (0.3–)0.5–1.1(–1.4) mm in diameter, immersed, becoming adnate with age; proper margin black, thin, ± persistent, excluded with age; disc black, usually with a fine white pruina, plane, becoming slightly convex with age; exciple narrow, poorly differentiated, of the aethalea-type (Fig. 48C) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), often reduced, similar in structure and orientation to the paraphyses, transient with the deep reddish brown hypothecium (leptoclinoides-brown, textura intricata), outer excipular hyphae parallel, moderately 383 swollen (textura oblita) and moderately carbonized with various amounts of brown and aeruginose pigments (cf. elachista-brown & cinereorufa-green), pigmentation ± continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachistabrown) and a diffuse, aeruginose pigment (HNO3+ violet, cinereorufa-green). Asci — 8spored, clavate, Bacidia-type. ASCOSPORES (Fig. 48E–I) — oblong to narrowly oblong, very rarely constricted, with obtuse ends, not curved, (13.0–)16.3–[18.6]–20.9(–25.0) x (6.0–)6.2–[7.1]–8.0(–10.0) µm (n = 60); one-septate, proper septum briefly thickened but very soon becoming reduced during spore ontogeny, lateral wall thickenings absent (± Physconia-type); ornamentation microrugulate (to faintly striate); septum with septal pore canal, simple pore and undifferentiated pore plug; spore wall (Fig. 48G) differentiated into smooth to indistinctly cracked, thick perispore (0.30–0.40 µm), narrow intermediate layer (< 0.03 µm), thick proper spore wall (0.35–0.53 µm) and moderately thickened endospore (0.18–0.30 µm). Pycnidia rare, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 3.0–4.5x 1.0–1.5 µm (20). Chemistry.—Typically with the depside atranorin and traces of the depsidone norstictic acid. Several xanthones have also been detected with HPLC by J. Elix: 4, 5dichloronorlichexanthone, arthothelin, thiophanic acid. However, the thallus does not react C+ orange and the UV-fluorescence is only faint, possibly obscured by the large 384 amount of Ca-oxalate pruina on the thallus surface. Thalli usually react K+ yellow, rarely K+ yellow-red (forming orange, needle-shaped crystals if observed in the compound microscope), P ± yellow, C– , KC–, CK–. UV± pale yellow to beige. Thallus and apothecia react strongly amyloid. Substrate and ecology.—On limestone (HCl+ strongly reacting) or sandstone with some traces of carbonates (HCl+ weakly reacting, rarely HCl–). Distribution (Fig. 9).—Currently known only from the U.S.A. (Utah, New Mexico, Colorado, Northern Arizona, California). Notes.—The thick, pruinose, areolate to sublobate, deeply amyloid thalli of B. navajoensis are very distinct and specimens are not easily confused with other species. Type specimens collected by Nebecker were originally identified as B. retrovertens but do not resemble that species. Although the species is very distinct, only a few specimens have so far been collected. Representative specimens examined.––UNITED STATES. ARIZONA Apache Co. Nash 9111 (ASU). CALIFORNIA San Bernardino Co. Nash 10350 (ASU). COLORADO Larimer Co. Ryan 12132 (ASU).NEW MEXICO San Juan Co. Marsh 12 (ASU). UTAH Grand Co. Ryan 11636 (ASU). UTAH Kane Co. Nebecker 1581, 1574, 1580 (ASU). BUELLIA REGINEAE Bungartz sp. nov. Thallus saxicolus, crustaceus, areolatus vel bullato-areolatus, crassus, eburneus, cum hypothallo atero. Apothecia sessilia, lecideina, marginibus propriis tenuis. Excipulum tenuis, fulvum, sine pigmento aeruginoso, carbonaceum. Asci 8-spori. Sporae uniseptae, 385 oblongae vel ellipsoideae, septibus incrassatus 11–19 x 5.5-10 µm. Pycnidia globosa. Conidia bacilliformes, 4-5 x 0.5-1.0 µm. Thallus atranorina, gangaleoidina, diploicina, fulgidina et acida haematommica et virensica continens. Medulla amyloideus. Etymology.—The species is dedicated to my sister Regine. The species epithet “regineae” also reminds of the superficially similar but not closely related Buellia capitis-regum W.A. Weber, which has a similarly “gnarled” thallus surface. TYPE: MEXICO. BAJA CALIFORNIA. Isla de Guadalupe, along west coast at "Fondeadero del Oeste" near light, 28º58'50"N, 118º18'50"W, on N-facing cliffs above the ocean, on basalt, ca. 80 m alt., 1 January 1996, Nash 38303 (ASU–holotype designated here!). Thallus (Fig. 49A,B).—crustose, distinctly thickened, ± continuous, epilithic; bullateareolate, thallus surface “gnarled”; prothallus distinct, delimiting the thallus as a black outline; thallus surface smooth (but often eroded in herbarium specimens), matt or ± shiny, usually white to pale ivory, rarely whitish gray, epruinose, phenocorticate; lacking crystals within thallus medulla (H2SO4–). Apothecia lecideine, (0.3–)0.6–1.1(–1.4) mm in diameter, soon sessile; proper margin thin, black, rarely persistent, soon excluded with age; disc black, epruinose, plane, sometimes ± convex with age; proper exciple of the dispersa-type (Fig. 49C) sensu Scheidegger (1993), i.e. inner exciple distinct, not reduced, hyphae pigmented, prosoplectenchymatous (textura oblita), extending from the deep reddish brown hypothecium (leptoclinoides-brown, textura intricata); outer excipular hyphae short-celled, cells angular, distinctly swollen (textura angularis) and 386 usually ± carbonized with various amounts of a brown pigment (cf. elachista-brown), pigmentation continuous with the epihymenium; hymenium hyaline, inspersed with oil droplets; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown). Asci 8-spored, clavate, Bacidia-type. Ascospores (Fig. 49D–F) oblong to ellipsoid, usually not constricted, with obtuse ends, not curved, (11.0– )12.8–[14.6]–16.3(–19.0) x (5.5–)6.4–[7.4]–8.4(–10.0) µm (n = 60); one-septate, proper septum distinctly and ± persistently thickened during spore ontogeny, lateral wall inconspiciously thickened (± Callispora-type); ornamentation absent (not visible in DIC); septum with septal pore canal, simple pore and undifferentiated pore plug; spore wall (Fig. 49F) differentiated into smooth, thin perispore (0.06–0.09 µm), indistinct intermediate layer (< 0.01 µm), moderately thickened proper spore wall (0.26–0.30 µm) and thick endospore (0.23–0.41 µm). Pycnidia infrequent, globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 4.0–5.0x 0.5–1.0 µm (20). Chemistry.—The depsides atranorin and chloroatranorin together with various artifacts from hydrolysis of atranorin (chlorohaematommic acid, haematommic acid, methyl-βorsellinate) have been detected. Other biosynthetically related depsides found in the specimens are: gangaleoidin, norgangaleoidin, chlorolecidioidin, dechlorogangaleoidin. One specimen contains brialmontin 1. The following biosynthetically related depsidones were detected: diploicin, 3-dechlorodiploicin, fulgidin and isofulgidin. One specimen 387 contains virensic acid. (HPLC by J. Elix, Canberra). K+ deep yellow, P+ faintly yellow, C–, KC–, CK–. UV+ bright yellow. The thallus medulla and the apothecia react strongly amyloid. Substrate and ecology.—Growing on siliceous mineral-poor coastal rock (generally HCl–). Distribution (Fig. 9).—Currently known only from the coast of Baja California and southern California. Notes.—Buellia regineae is closely related to a group of species, which are commonly treated within Hafellia, a genus described by Kalb (1986) to accommodate species with an oil inspersed hymenium and conspicuous lateral spore wall thickenings. Etayo & Marbach (2003, p. 373) also emphasize that all species of Hafellia have “… strongly branched paraphyses (forming an epithecium) …” and typically occur in habitats with high humidity. Most species included in the genus are corticolous. Hafellia thus appears very distinct from species related to Buellia aethalea, which is often regarded as the core group of the genus Buellia. Unfortunately this concept is currently in direct violation of the Botanical Code as adopted in 1999 in St. Louis (Greuter et al. 2000). This currently accepted code lists Buellia disciformis as the type of the conserved genus Buellia. The listed type clearly belongs to Hafellia and was included within that genus by Marbach & Mayrhofer (in Marbach 2000). This transfer is formally invalid because a listed type of one genus (i.e. Buellia) cannot belong to another genus (i.e. Hafellia). To avoid taxonomic upheaval Moberg et al. (1999) proposed to change the listed type from B. disciformis to B. 388 aethalea. The majority of lichen taxonomists readily accept the genus Hafellia (Kalb 1986; Birkbeck et al. 1990; Scheidegger 1991; Sheard 1992; Scheidegger 1993; Pusswald et al. 1994; Sheard & Tønsberg 1995; Marbach 2000; Hafellner & Türk 2001; Eriksson et al. 2002a; Etayo & Marbach 2003). In contrast, Nordin (2000) adopted a very broad concept of Buellia suggesting that the proposal to change the listed type will most likely be rejected. Rico et al. (2003) went even further arguing that the acceptance of the most recent code implies that Hafellia must be treated as a synonym of Buellia s.str. Until a decision on the proposal submitted by Moberg et al. (1999) has formally been reached, species with affinities to Hafellia must therefore be included within Buellia s.l. Buellia regineae is closely related to B. leptoclinoides (Nyl.) J. Steiner (= Hafellia leptoclinoides (Nyl.) Scheid. & H. Mayrhofer nom. illegit.). Material of B. leptoclinoides was examined for comparison from M and the private herbarium of C. Scheidegger. The specimens have a more distinct lateral wall thickening than B. regineae. Nevertheless both species are very similar but the medulla of Buellia regineae reacts strongly amyloid and specimens contain fulgidin, isofulgidin and diploicin instead of placodiolic acid. Both species are saxicolous, which is unusual given that the majority of related species appears to be corticolous. Paraphyses of both B. regineae and B. leptoclinoides are not considerably different from other saxicolous species and a distinct layer on the surface of the hymenium could not be observed (i.e. a distinct epithecium is absent). Thalli growing close to the seashore and subject to spray often have more exuberant areoles and look “gnarled”. 389 Representative specimens examined.––MEXICO. BAJA CALIFORNIA. Nash 26092, 26093, 38303 (ASU); Weber s.n. (COLO). Specimens of B. leptoclinoides examined for comparison.––FRANCE. EASTERN PYRENEES (= Département Pyrenées Orientale). Scheidegger 8480a,b (hb. Scheidegger); Kalb Lichenes Neotropici Exs. 372 (M-0061356). ALSACE (= Département Alsace). Suza s.n. (M-0061354). EUROPE. UNKNOWN LOCALITY. Arnold s.n. (M-0051355). Comment.—A specimen selected by Meyer & Printzen (2000) as a reference collection for the pigment leptoclinoides-brown (Vezda Lichenes Selecti Exs. 446; M0061353) has been erroneously identified as H. leptoclinoides. The specimen in M belongs to B. saxorum. The same pigment is nevertheless present in this specimen and it is unnecessary to change the reference collection for this pigment. BUELLIA SHEARDII Bungartz, sp. nov. Thallus saxicolus, crustaceus, areolatus, tenuis vel crassus, eburneus, cum hypothallo atero. Apothecia sessilia, lecideina, marginibus propriis crassis. Excipulum crassum, fulvo-caeruleum, pigmentum aeruginosum continens, carbonaceum. Asci 8-spori. Sporae uniseptae, oblongae vel ellipsoideae, septibus angustus, 8–13.5 x 4-6 µm. Pycnidia globosa. Conidia bacilliformes vel fusiformes, 6-9 x 1.0-1.5 µm. Thallus acida norstictica et connorstictica continens. Medulla amyloideus. Etymology.—The species is named to honor Dr. John W. Sheard for his outstanding contributions to the taxonomy of Rinodina and Buellia. 390 TYPE: MEXICO. SINALOA. Topolobampo, near Gulf of California on north-facing hillside, on basalt, ca. 30 m alt., 20 March 1975, Nash 10086 (ASU–holotype designated here!, one isotype at MEXU, one isotype at US). Thallus (Fig. 50A,B) crustose, thin to moderately thickened, ± continuous, epilithic; areolate; prothallus distinct, delimiting the thallus as a black outline, rarely also between the areoles; thallus surface matt and dull, usually ivory, rarely pale beige, pruinose, phenocorticate; entire thallus with large mineral crystals (ca. 40-60 µm in diameter; H2SO4–, not forming needles). Apothecia lecideine, (0.2–)0.3–0.6(–0.8) mm in diameter, soon adnate to sessile; proper margin prominent, black or rarely masked by grayish remains of necrotic thalline material (thalline veil), ± persistent, rarely excluded with age; disc black, epruinose or faintly pruinose, plane, becoming only slightly convex with age; proper exciple of the dispersa-type (Fig. 50B) sensu Scheidegger (1993), i.e. inner excipular hyphae distinct, not reduced, pigmented, prosoplectenchymatous (textura oblita), extending from the deep reddish brown hypothecium (leptoclinoides-brown, textura intricata); outer excipular hyphae short-celled, cells angular, distinctly swollen (textura angularis) and usually ± carbonized with various amounts of brown and aeruginose pigments (cf. elachista-brown and cinereorufa-green, HNO3+ violet), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown) and a diffuse aeruginose pigment (HNO3+ violet, cinereorufagreen). Asci (Fig. 50C) 8-spored, clavate, Bacidia-type. Ascospores (Fig. 50C) oblong to 391 ellipsoid, usually not constricted, with obtuse ends, not curved, (8.0–)8.8–[10.2]– 11.6(–13.5) x (4.0–)4.1–[4.8]–5.5(–6.0) µm (n = 39); one-septate, proper septum narrow, not thickening during spore ontogeny, lateral wall thickenings absent [Beltraminea (=Buellia)-type]; ornamentation absent (not visible in DIC). Pycnidia rare, globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform to fusiform, 6.0–9.0 x 1.0–1.5 µm (20). Chemistry.—With the biosynthetically related depsides atranorin and chloroatranorin and the depsidones norstictic and connorstictic. Thallus usually K+ yellow to red (orange, needle-shaped crystals forming in the compound microscope), P– or + yellow, C–, KC–, CK–. UV– (dark). The thallus and apothecia react amyloid. Substrate and ecology.—Growing on siliceous mineral-poor coastal rock (generally HCl–). Distribution (Fig. 10).—Currently known only from the coast of Sinaloa, along the Gulf of California (Sea of Cortez). Notes—Because of the plane apothecia with a ± persistent margin B. sheardii superficially resembles B. disciformis, which, however, has a rimose-areolate rather than a distinctly areolate thallus, ascospores with median septum thickenings and does not contain cinereorufa-green in the exciple. 392 Additional specimen examined.––MEXICO. SINALOA. 39 km N of Los Mochis along route 15; 26°07'00''N, 109°03'00''W; 60 m, Nash 12088 (ASU). BUELLIA TYROLENSIS Körb., Parerg. Lich.: 187. 1860. TYPE: ITALY. TRENTINO (= SOUTH TIROL). Naifthale bei Meran, ca. 46°40'N, 11°09'E, 1853, Bamberger ex h. Heufl. s.n. (UPS–isotype). Taxonomic note.—The “Parerga Lichenologica” was issued in several "Lieferungen" beginning with pages 1-96 in 1859. Pages 97-192 were first published in the following year (1860) with the species description of Buellia tyrolensis on page 187. Zahlbruckner (1931) and Scheidegger (1993) both use the spelling "tirolensis" and cite page 460 instead of page 187. However, on page 460 (first published in 1865) no species of the genus Buellia is treated. According to Articles 60.1 and 60.4 of the code (Greuter et al. 2000) the original spelling with "y" as Buellia tyrolensis is to be maintained and must not be changed to "tirolensis". Buellia buellioides (Metzler ex Arnold) Buschardt, Bibl. Lich. 10: 86. 1979.— Rinodina buellioides Metzler in Arnold, Verh. zool. bot. Ges. Wien 23: 112 (1873) comb. novum pro Buellia fusca sensu Arnold nom. illegit., Verh. zool. bot Ges. Wien 22: 291. 1872. TYPE: ITALY. TRENTINO (= SOUTH TIROL). On porphyric rocks above Gries near Bozen, 46°31'N, 11°19'60''E [original label data: An Porhyrfelsen oberhalb Gries bei Bozen in Südtirol], 29 August 1872, Arnold Exs. 495 (M-0061319–lectotype selected here!; M-0061317–isoletotype!). Note: Buschardt (1979) did not specify a type from 393 Arnold’s exsiccati collection and the specimen with the barcode M-0061319 is therefore selected here as the lectotype. Buellia fusca (Anzi) Kernst., Zeitschr. Ferdinandeums 35: 306. 1893.—Buellia spuria var. fusca Anzi, Cal. Lich. Sondr.: 87 (1860). TYPE: ITALY. LOMBARDIA. On silicious rock near Comun Nuovo (San Martino), 46°46'60''N, 11°13'00'' E [original label data: Ad rupes silaceas prope Novum-Comum (S. Martino)], Anzi s.n. [Lich. Lang. no. 195] (M0061318!, BERN, UPS, W–isotypes) Buellia novomexicana de Lesd., Ann. Crypt. Exot. 5(2): 128 (1932). TYPE: not found, probably destroyed in Word War II. Buellia novomexicana f. pruinosa de Lesd., Ann. Crypt. Exot. 5(2): 129 (1932). TYPE: U.S.A. NEW MEXICO. San Miguel Co. Near Las Vegas. Gallinas South Canyon, 34°04' N 107°01' W, on silicious rocks, 1870 m [original label data: New Mexico. Environs de Las Vegas: Canon Sud, sur roches silicieuses], 14 April 1927, Arsène Brouard s.n. (ex hb. Vězda, STU–lectotype selected here!). Note: Type material of forma pruinosa from STU was the only material available. This material is selected here as the lectotype because de Lesdain’s holotype collection, originally located in Dunkerque (France), was probably destroyed during the Second World War. Buellia zapotensis de Lesd., Lichens du Mexique: 26. 1914. TYPE: MEXICO. MICHOACÁN. Morelia. Small hills west of Zapote hill, 19°41'60''N, 101°07'00''W, 1900 m [original label data: Morelia. lomas à l’ouest de la loma del Zapote, 1900 m], 2 February 1910, Arsène Brouard 3701 (US–lectotype selected here!). Note: The lectotype specimen 394 selected here from a duplicate of the original collection, has a non amyloid medulla (I– ) contrary to the protologue. For additional not verified synonyms see Scheidegger (1993). Thallus (Fig. 51A,B) crustose, thin to moderately thickened, ± continuous, epilithic; areolate; prothallus conspiciously black, in most specimens strongly developed and growing between the areoles (forming a hypothallus), rarely only surrounding the thallus outline; thallus surface matt and smooth, rarely ± shiny, usually deep brown, rarely olive brown, epruinose, phenocorticate; lacking crystals within the thallus medulla (H2SO4–). Apothecia lecideine, (0.2–)0.3–0.4(–0.5) mm in diameter, remaining immersed to indistinctly adnate; proper margin prominent, black or masked by grayish remains of necrotic thalline material (thalline veil), usually persistent, rarely excluded with age; disc black, epruinose, rarely with a faint white pruina, plane, rarely becoming slightly convex with age; proper exciple narrow, poorly differentiated, of the aethalea-type (Fig. 51C) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), often reduced, similar in structure and orientation to the paraphyses, transient with the deep reddish brown hypothecium (leptoclinoides-brown, textura intricata), outer excipular hyphae parallel, moderately swollen (textura oblita) and usually strongly carbonized with various amounts of a brown pigment (cf. elachista-brown), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown). Asci 8-spored, clavate, Bacidia- 395 type. Ascospores (Fig. 51D,E) oblong to ellipsoid, usually not constricted, with obtuse ends, not curved, (9.0–)9.4–[10.4]–11.3(–15.0) x (5.5–)5.8–[6.2]–6.7(–8.0) µm (n = 60); one-septate, proper septum narrow, not thickened during spore ontogeny, lateral wall thickenings absent [± Beltraminea (=Buellia)-type]; ornamentation absent (not visible in DIC); septum with septal pore canal, simple pore and undifferentiated pore plug; spore wall (Fig. 51E) differentiated into smooth, thin perispore (0.08–0.15 µm), very narrow intermediate layer (< 0.20 µm), thick proper spore wall (0.25–0.50 µm) and thick endospore (0.16–0.68 µm). Pycnidia rare to common, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogenous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 4.5–7.0x 0.5–1.0 µm (n = 20). Chemistry.— The thalli contain the biosynthetically related depsidones norstictic and connorstictic acid and/or the depside 2'-O-methylperlatolic acid. Spot test reactions are not distinct on the dark thallus and must be confirmed in the compound microscope. K+ yellow to red (± forming orange, needle-shaped crystals) or K– (no crystals), P– or + faintly yellow, C–, KC–, CK–. UV– (dark). The thallus is not amyloid, but apothecia react amyloid in Lugol’s (thallus reactions of the dark thallus should be confirmed in the compound microscope). Substrate and ecology.—On a wide variety of silicieous (HCL-) rock, typically on ± shaded and sheltered boulders and cliffs. 396 Distribution (Fig. 11).—Common and widely distributed in montane areas of the Greater Sonoran Desert Region. The species is also known from Mediterranean Europe, where it appears to be quite rare. Notes.—Generally no distinct thickening of the spore septum has been observed in any of the specimens examined with the light microscope. Scheidegger (1993) therefore assigned the spores to the Buellia-type. Unfortunately these observations do not agree very well with specimens examined with the TEM. In the TEM the endospore appears considerably thickened especially along the spore septum. Currently not enough material has been examined to evaluate this discrepancy between light and electron microscopy. Scheidegger (1993) distinguished B. fusca (with 2'-O-methylperlatolic acid) from B. tyrolensis (with norstictic acid). Although the majority of specimens examined can be divided into these two chemotypes, some specimens with both norstictic and 2'-Omethylperlatolic acid have been found. All chemotypes (only norstictic, only 2'-Omethylperlatolic and specimens containing both substances) are morphologically and anatomically identical and show no distinctly different distribution patterns. This situation is surprisingly different from B. stellulata (with 2'-O-methylperlatolic acid, coastal, non amyloid medulla) and B. spuria (with norstictic acid, inland, amyloid medulla), where the same lichen substances clearly denote two distinctly different species. Representative specimens examined.––EUROPE. ITALY. TRENTINO (= SOUTH TIROL). Arnold s.n. (M-0061312, M-0061314, M-0061316); Doppelbaur 10288 (M0061320); Metzler s.n. (M-0061313, M-0061321). LIGURIA. Sbarbaro s.n. (MSC-40629, 397 M-0061315).––NORTH AMERICA. MEXICO. BAJA CALIFORNIA SUR. Nash 12758, 39833, 40016 (ASU). CHIHUAHUA. Nash 31198, 36099, 36545, 36677, 37521, 37837, 13508b, 13720 (ASU). SONORA. Nash 25500, 25611, 37942, 12519 (ASU); Nash 11959 (MSC-335429). . –– SPAIN. ALMERIA. Scheidegger Inv. Nr. 8640 (hb. Scheidegger). UNITED STATES. ARIZONA. Cochise Co. Hertel 40053a (M); Lehto s.n., Nash 3703, 3740, 3742, 3821, Ryan 10961, 10978, 10782 (ASU); Weber s.n., S8782, S8782, Exs. 408 (COLO); Weber 8782 (MSC-40696); Shushan 8782 (MSC-40696); Shushan S-8782 (M0061367). Gila Co. Nash 7400b (ASU). Graham Co. Nash 36043 (ASU). Pima Co. Darrow 1448 (ASU); Hertel 39915 (M-0061399); Nash 27406a, 4090, 4146, Ryan 20310 (ASU). Santa Cruz Co. Darrow 477, 821, Nash 25242, 25243, 25355, 25774, 13011, 7161 (ASU); Scheidegger Inv. Nr. 31-39 (hb. Scheidegger); Schramm 196, Zschau s.n. (ASU). COLORADO. Colorado Co. Shushan 8185 (MSC-40707). MISSOURI. Taney Co. Wetmore 69088 (MIN). NEW MEXICO. Doña Ana Co. Nash 7922 (ASU). San Miguel Co. Nash 16048 (ASU). Sierra Co. Nash 7116 (ASU). Soccoro Co. Shushan S-6932 (M-0061365). SOUTH DAKOTA. Custer Co. Wetmore 10175 (MSC-70750). TEXAS. Brewster Co. Nash 15037a (ASU); Wetmore 18488b, 19391 (MIN, M-00613161); Anderson S-18696 (M0061366); Wetmore 19417 (MSC-374082); Anderson 18696 (MSC-45835); Shushan 18696 (MSC-45835). BUELLIA VILIS Th. Fr., Kgl. Vetensk. Akad. Handl. 7(2): 44. 1867. TYPE: NORWAY. SVALBARD (= SPITSBERGEN). On western sea-shore rocks. [original label data: ad saxa litore occidentali], Nordenskjöld s.n. (UPS?—type not seen). 398 Taxonomic note.—Type material of Th. Fries at UPS may be lost and a neotype may have to be selected. Scheidegger (1993) also did not see the type. The protologue and the descriptions in Leighton (1868) and Steiner (1907) are, however, very detailed and it can be reasonably assumed that the type shows the same characters as described there. A specimen from Körber’s “Typenherbar” in L is annotated as B. vilis (L-0065286). This specimen, however has hyaline, non-septate ascospores and possibly belongs to Lecidella. Buellia vilis is not closely related to B. notabilis Lynge even though the protologue and a description of B. notabilis by Thomson (1997a) suggest otherwise. A “kleptotype” specimen of B. notabilis at MSC has a distinctly epilithic, areolate, gray thallus with a non-amyloid medulla (I-). The lecideine apothecia of this specimen are sunken between the areoles. Because of a Lecanora-type ascus and hyaline hypothecium, B. notabilis may better be placed into Rinodina. For synonyms of B. vilis see Scheidegger (1993). Thallus (Fig. 52) crustose, thin, discontinuous, hidden, chasmolithic; i. e. forming inconspicuous, poorly delimited granules between the mineral grains of the substrate; prothallus absent; thallus surface matt and dull, not shiny, usually pale gray, rarely pale brown, epruinose, phenocorticate; lacking crystals within thallus medulla (H2SO4–). Apothecia lecideine, (0.2–)0.5–1.1(–1.2) mm in diameter, soon sessile; proper margin thin, black, usually persistent, rarely excluded with age; disc black, epruinose, plane, ± becoming slightly convex with age; proper exciple of the vilis-type (Fig. 52C, D) sensu 399 Scheidegger (1993), i.e. inner excipular hyphae hyaline (reacting strongly I+ blue), prosoplectenchymatous, loosely interwoven (textura intricata), transient with the hyaline hypothecium (reacting strongly I+ blue, textura intricata), outer excipular hyphae parallel, thin (textura oblita) and usually very strongly carbonized with large amounts of a dull brown-red to blackish pigment (atra-red, HNO3+ deep purple), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a deep brown to black pigment cap (atrared, HNO3+ deep purple). Asci 8-spored, clavate, Bacidia-type. Ascospores oblong to ellipsoid, usually constricted (with age), with obtuse ends, not curved, (12.0–)13.1– [14.5]–15.9(–18.0) x (5.0–)5.7–[6.6]–7.4(–9.0) µm (n = 60); one-septate, proper septum narrow, not thickened during spore ontogeny, lateral wall thickenings absent [Beltraminea (=Buellia)-type]; ornamentation absent (not visible in DIC). Pycnidia very rare, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 4.5–7.0x 0.5–1.0 µm (n = 20). Chemistry.—No substances found with TLC. All spot tests negative (K–, P–, C–, KC–, CK–). UV– (dark). The thallus hyphae react very strongly amyloid like the hyphae in the apothecium (even if tested with low iodine concentrations!). 400 Substrate and ecology.—According to Scheidegger (1993) typicallly found on small pebbles in exposed habitats. The specimens from Arizona and New Mexico were found on sandstone. Distribution (Fig. 12).—The species appears to be extremely rare in the Sonoran Desert Region and is probably restricted to montane to subalpine or even alpine habitats. A single record is currently known from the Mogollon Rim in Arizona. A second specimen, deposited at ASU has been collected at 1768 m elevation outside the Sonoran Region in New Mexico (Rankert 116). Notes.—Buellia vilis has frequently been reported from the North American Southwest but almost all reports are erroneous and based on misidentifications of B. sequax (Nyl.) Zahlbr. (see Bungartz et al. 2004b). Until recently, no single specimen was known from the area (Bungartz & Nash III 2004b). Buellia vilis is very isolated even within an expanded genus concept of Buellia s.l. because of the unique exciple type characterized by a pigment not found in any other species. The species may best be placed in a monotypic genus but the genus concept in Buellia is currently not well resolved and the introduction of new genera may only contribute to further confusion. Representative specimens examined.––AUSTRIA. TIROL. Arnold s.n. (M-0061300). FRANCE. Hautes Pyrenées. Scheidegger Inv. Nr. 8056, 8383, 8498, 8501 (hb. Scheidegger). ITALY. TRENTINO (= SOUTH TIROL). Arnold s.n. (M-0061301, M0061302). UNITED STATES. ALASKA. North Slope Co. Fryday 8279 (MSC). ARIZONA. Cocinino Co. Hertel 40242 (M). NEW MEXICO. San Juan Co. Rankert 116 (ASU). 401 TABLE 7. References to detailed descriptions of all species known from the Sonoran Desert Region, including recent synonyms. accepted name B. aethalea B. argillicola B. badia B. christophii B. concinna B. dispersa B. eganii B. halonia B. lacteoidea B. lepidastroidea B. mamillana B. nashii B. navajoensis B. paniformis B. prospersa B. pullata B. regineae B. ryanii B. sequax B. sheardii B. spuria B. stellulata B. subaethalea B. subalbula B. subdisciformis B. tergua B. tesserata B. trachyspora B. tyrolensis B. uberior B. vilis recent synonym(s) B. turgescens B. semitensis B. retrovertens B. glaziouana, B. thomae Amandinea lecideina B. saxicola B. maritima B. fimbriata, B. cerussata B. novomexicana, B. fusca references Bungartz & Nash (2004c). The Bryologist, submitted. current article Bungartz & Nash (2004b). The Bryologist 107: 21-27. Bungartz et al. (2004b). Can. J. Bot., in press. Bungartz et al. (2004c). The Bryologist 107: 21-27. Bungartz et al. (2002) Bibl. Lich. 82: 19-35. Bungartz & Nash (2004c). The Bryologist, submitted. Bungartz et al. (2004c). The Bryologist, submitted. Bungartz & Nash (2004c). The Bryologist, submitted. current article Bungartz et al. (2004c). The Bryologist 107: 21-27, Weber (1986). Mycotaxon 27:451-497. current article current article Weber (1971). The Bryologist 74:185-191. Bungartz et al. (2004b). Can. J. Bot. (in press) Bungartz et al. (2004b). Can. J. Bot. (in press) current article Bungartz et al. (2004b). Can. J. Bot. (in press) Bungartz et al. (2004b). Can. J. Bot. (in press), Scheidegger (1993). Lichenologist 25:315-364. current article Bungartz & Nash (2004c). The Bryologist, submitted Bungartz & Nash (2004c). The Bryologist, submitted Bungartz et al. (2004c). The Bryologist, submitted Bungartz & Nash (2004a) Bibl. Lich. 88:49-66. Scheidegger (1993). Lichenologist 25:315-364. Bungartz et al. (2004b). Can. J. Bot. (in press) Rico et al. (2003). Lichenologist 35:117-124 Bungartz et al. (2004c). The Bryologist 107: 21-27. current article Scheidegger (1993). The Lichenologist 25:315-364, Scheidegger (1987). Botanica Helvetica 97:99-116. current article 402 → FIGURE 45. Light micrographs of Buellia argillicola (Nash 12152).—A. Overview of the rimose thallus.—B. Close-up of the rimose thallus and apothecia.—C. Cross section of the aethalea-type exciple.—D. Oblong, ascospores.—E. Ascospore: no septum thickening is present during the spore ontogeny. 403 404 → FIGURE 46. Buellia lepidastroidea (A–E. light micrographs; F–H. TEM micrographs).—A. Overview of the areolate, sublobate thallus with sessile apothecia (Nash 6742a).—B. Close-up of thallus and apothecia (Nash 6742a).—C. Cross section through the apothecium with inspers hymenium (Nash 34458).—D. Premature ascospore with distinct septal thickening (Nash 34458).—E. Conidia (Nash 34458).—F. Bacidiatype ascus (Nash 38438; for designation of the different layers see Bellemère 1994): The a- and b-layer (ab) are barely visible and cannot reliably be distinguished (possibly as a result of fixation artifacts); (c) outer electron opaque c-layer; (d1) d1-layer, i.e. the outer tholus, which is distinctly laminated (in light microscopy this outer part stains deep blue with Lugol’s iodine); (d2) d2-layer, i.e. the inner tholus, which is not layered and ± homogeneous (not staining in Lugol’s iodine); (oc) ocular chamber .—G. Mature ascospore with reduced septal thickenings (Nash 38438).—H. Spore wall of a mature ascospore (Nash 38438): (as) ascus wall; (s) mucilaginous sheath; (1) perispore; (2) intermediate layer; (3) proper spore wall; (4) endospore. 405 406 → FIGURE 47. Buellia nashii.—A. Subsquamulose thallus with sessile apothecia (Nash 6724a).—B. Close-up of thallus (Nash 6724a).—C. Cross section of an apothecium with dispersa-type exciple (Nash 15313b).—D. Overmature, slightly constricted ascospore with reduced septum thickening (Nash 15313b).—E. Microrugulate ascospore ornamentation visible at a late stage of the spore ontogeny (Nash 15313b).—F. Conidia (Nash 15313b). 407 408 → FIGURE 48. Buellia navajoensis.—A. Overview of areolate thallus (Nebecker 1581). — B. Close-up of areolate thallus (Nebecker 1581).—C. Cross section of an apothecium with aethalea-type exciple (Nebecker 1581).—D. Hymenium with asci, paraphyses and one ascospore (Nash 1581).—E. Mature ascospore: an indistinct septum can be seen in some of the spores (Nash 1581). — F. Immature ascospore prior to septum formation (Nash 1577–holotype).—G. Mature ascospore (Nash 1577–holotype).—H. Wall layers in an immature ascospore: (1) outer layer (perispore and mucilaginous sheath not distinctly differentiated); (2) proper spore wall and (3) endospore beginning to become differentiated (Nash 1577–holotype).—I. Wall layers in a mature ascospore: (as) ascus wall; (s) mucilaginous sheath; (1) perispore; (2) intermediate layer; (3) proper spore wall; (4) endospore (Nash 1577–holotype). 409 410 → FIGURE 49. Buellia regineae.—A. Overview of the bullate-areolate thallus outlined by a distinct black prothallus (Wetmore 75787).—B. Close-up of the thallus (Wetmore 75787).—C. Apothecium with inspersed hymenium and dispersa-type exciple (Wetmore 63682).—D. Mature ascospore with indistinct lateral wall thickenings (arrow; Weber Exs. 89).—E. Mature ascospore (Nash 34319).—F. Wall layers in a mature ascospore: (s) mucilaginous sheath; (1) perispore; (2) indistinct, narrow intermediate layer; (3) proper spore wall; (4) endospore (Nash 34319). 411 412 → FIGURE 50. Buellia sheardii (Nash 10086 – holotype).—A. Areolate thallus with sessile apothecia and a black hypothallus.—B. Cross section of an apothecium with dispersa-type exciple.—C. Hymenium with Bacidia-type ascus (the arrow indicates the characteristic staining of the tholus flanks in Lugol’s iodine). 413 414 → FIGURE 51. Buellia tyrolensis.—A. Overview of the areolate thallus with black hypothallus (Nash 10987).—B. Close-up of the thallus (Nash 10987).—C. Cross section with aethalea-type exciple (Ryan 10987).—D. Mature ascospore (Bungartz 1575): The endospore appears considerably thickened in the TEM at this stage of the spore ontogeny although no septal thickening is visible in the light microscope.—E. Wall layers in a mature ascospore: (1) perispore; (2) indistinct, narrow intermediate layer; (3) proper spore wall; (4) endospore (Bungartz 1575). 415 416 → FIGURE 52. Buellia vilis.—A. Overview of the chasmolithic thallus (Hertel 40242).— B. Close-up of chasmolithic thallus (Hertel 40242).—C. cross section of an apothecium (Marsh 116).—D. cross section of an apothecium with vilis-type exciple (Marsh 116). 417 418 KEY Saxicolous species of Buellia s.l. with one-septate ascospores in the Sonoran Desert Region. 1 Thallus parasitic on other lichens ......................................................................... 2 Thallus not parasitic ............................................................................................. 3 2(1) Thallus deep brown, bullate-areolate to distinctly squamulose, parasitic on a variety of lichen genera including Xanthoparmelia, Acarospora and Dimelaena, medulla KC–, C– (without secondary metabolites or traces from the host lichen), spore ornamentation indistinct ....................................... B. badia (Fr.) A. Massal. Thallus light gray to dark gray, areolate, forming distinct insular patches in Schaereria fuscocinerea, medulla KC+ fleeting pink, C+ fleeting pink (medulla with gyrophoric acid), spore ornamentation striate ..................... B. uberior Anzi 3(1) Thallus C+ orange, KC+ orange (with xanthones) .............................................. 4 Thallus C–, KC– (with or without xanthones) .................................................... 5 4(3) Thallus granular to minutely bullate, not areolate; prothallus absent; spores with ± tapered ends, frequently curved and crescent-shaped, rarely not curved and ± citriform; exciple brown (HNO3–); montane to subalpine .................................... ................................................................................................ B. concinna Th. Fr. Thallus thick, areolate, usually with a distinct black prothallus along the margin; spores ellipsoid to oblong, with obtuse ends, not curved; exiple aeruginose (HNO3+ violet); coastal .................................................. B. halonia (Ach.) Tuck. 5(3) Inner exciple distinctly paraplectenchymatous, formed by large, leptodermatous, ± isodiametric cells; outer exciple very strongly carbonized (small, globular cells 419 barely discernible, even in thin microtome sections); spores with a coarsely rugulate to areolate ornamentation (clearly visible at 400x); rare, subtropical; in the Sonoran Desert Region currently known only from Sinaloa and Chihuahua, Mexico ................................................................................. B. trachyspora Vain. Inner exciple not paraplectenchymatous, formed by mesodermatous hyphae; outer exciple moderately to strongly carbonized (individual cells discernible, at least in thin microtome sections); spores smooth or ornamented but not coarsely areolate (ornamentation, if present, barely visible at 400x); ................................ 6 6(5) Thallus with a chalky consistency, containing large amounts of Ca-oxalates (clusters of sulphate needles in H2SO4); cortex roughened, often strongly pruinose; substrate usually calcareous, (HCl+) .................................................... 7 Thallus not chalky, without Ca-oxalates (H2SO4–), cortex smooth but sometimes obscured by fine pruina; substrate usually not calcareous (HCl–) ..................... 11 7(6) Medulla amyloid (Iconc.+ blue) ............................................................................. 8 Medulla not amyloid (Iconc.–) ............................................................................... 9 8(7) Thallus thin, rimose (finely fissured), subeffigurate; bright white; without xanthones; apothecia with thick, persistent, lecideine margin; spores rugulate, oblong to ellipsoid; spore septum not thickening during ontogeny ........................ .......................................................................................................... B. argillicola Thallus thick, distinctly areolate (cracked) to sublobate; ivory, pale beige or with faint pinkish tinge; containing xanthones; apothecia with thin, ± reduced lecideine margin; spores microrugulate to faintly striate, oblong to narrowly 420 oblong, spore septum soon, but only briefly thickening during ontogeny ............. ........................................................................................................ B. navajoensis 9(7) Thallus rimose to rimose-areolate, ± continuous, usually delimited by distinctly blackened, rarely pale prothallus; apothecia immersed to adnate; exciple narrow, often reduced to few hyphae similar in structure and orientation to the paraphyses, end cells distinctly swollen, with fuscous brown pigment cap and diffuse, aeruginose pigment (HNO3+ violet), diffuse pigment extending across epihymenium; species restricted to coastal habitats ............................................... .............................................................................. B. subalbula (Nyl.) Müll. Arg. Thallus areolate to subsquamulose, rarely sublobate, ± continuous to dispersed, not delimited by prothallus; apothecia soon sessile; exciple distinct, strongly developed, inner hyphae pigmented, mesodermatous, extending from reddish brown hypothecium, outer hyphae moderately swollen with fuscous pigment cap, aeruginose pigment absent (HNO3–) or restricted to outermost exciple cells; widely distributed throughout the Sonoran Desert Region ............................... 10 10(9) Outermost exciple with small amounts of diffuse aeruginose pigment (HNO3+ violet); thallus usually K+ orange to red (orange, needle-shaped crystals), rarely K+ yellow or K– (no crystals); with norstictic and/or stictic acid .......... B. nashii Outermost exciple cells without diffuse aeruginose pigment (HNO3–); thallus K– or K+ yellow (no crystals); with 2'-O-methylperlatolic acid .................................. ............................................................................................ B. dispersa A. Massal. 421 (Note: A revision of this morphologically variable species group using both classical and molecular data is currently in progress. For a distinction of three morphotypes see Bungartz et al. 2002) 11(6) Thallus indistinct, chasmolithic to endolithic, dispersed granules hidden among mineral crystals or entirely hidden ..................................................................... 12 Thallus distinct, epilithic, establishing ± continuous on the substrate surface ... 14 12(11) Spores with ± persistent septum thickenings; thallus pale yellow or whitish, containing xanthones (UV± pale to bright yellow or orange); conidia filiform; restricted to coastal habitats ........................................ B. prospersa (Nyl.) Riddle (Note: Specimens of B. prospersa are usually distinctly epilithic but rarely have a poorly developed and ± discontinuous thallus, which may appear chasmolithic, see Bungartz et al. 2004a) Spores without septum thickening; thallus absent or white to pale gray, without xanthones (UV– or UV± pale but not bright yellow or orange); conidia bacilliform; not restricted to coastal habitats ..................................................... 13 13(12) Thallus endolithic to chasmolithic; hyphae strongly amyloid (I+ blackish blue); hypothecium and inner exciple hyaline, outer exciple strongly carbonized with blackish red pigment (HNO3+ deep purple); spores broadly oblong to ellipsoid; in the Sonoran Region currently known from a single locality (along the Mogollon Rim) ........................................................................................... B. vilis Thallus chasmolithic; hyphae not amyloid (concentrated I–); hypothecium deep reddish brown, outer exciple weekly carbonized with a brown pigment (HNO3–); 422 young spores narrowly oblong, becoming ellipsoid with age; common throughout the Sonoran Region ............................................................................... B. sequax 14(11) Hymenium inspersed with oil droplets (especially in KOH) ............................. 15 Hymenium not inspersed .................................................................................... 16 15(14) Medulla not amyloid (I–); thallus ± continuous to dispersed, apothecia small, usually < 0.8 mm (rarely up to 1.1 mm) in diameter; spores 7–17 x 5–8 µm, with distinct septum thickening but no lateral wall thickenings ........ B. lepidastroidea Medulla amyloid (I+ blue); thallus ± continuous, not becoming dispersed; apothecia large, up to 1.5 mm in diameter; spores 11–19 x 5.5–10 µm, with distinct septum thickening and inconspicuous lateral wall thickenings ................. ............................................................................................................. B. regineae 16(14) Thallus granular-areolate to verrucose; each spore cell covered by a broad, darkened band ................................................................. B. subaethalea de Lesd. Thallus not granular or verrucose, but sometimes bullate or subsquamulose to squamulose; spore cells not covered by darkened bands 17 17(16) Thallus areolate to squamulose or sublobate; apothecia soon sessile; hypothallus absent ................................................................................................................. 18 Thallus rimose to areolate, not subsquamulose or sublobate; apothecia immersed, becoming sessile with age; usually with a distinct, black hypothallus, rarely hypothallus absent ............................................................................................. 21 18(17) Thallus deep “chocolate” brown, epruinose; forming bullate to subsquamulose areoles or distinct squamules; juvenile thalli frequently parasitic, but becoming 423 independent with age; secondary metabolites absent (or traces from the host lichen) ...................................................................................................... B. badia Thallus not deep brown (ivory, pale brown, olive brown or dark gray); with or without pruina; never parasitic; with various secondary metabolites ................ 19 19(18) Thallus with diploicin, pale brown to tawny yellow (“isabelline”) and shiny, rarely with whitish pruina, forming strongly swollen, bullate areoles (“breadloaves”); epihymenium of immature apothecia shedding a thin epinecral layer when emerging through the thallus surface, exciple of young apothecia with a thin thalline collar, which is soon shed; coastal, rare (currently known only from Guadelupe Island and few localities on the Baja peninsula) .................................. .................................................................................... B. paniformis W.A. Weber Thallus without diploicin, areolate to subsquamulose or sublobate; epihymenium not covered by epinecral surface layer, not emerging with a thalline collar; common and widely distributed throughout the Sonoran Region ..................... 20 20(19) Outermost exciple always with small amounts of diffuse aeruginose pigment (HNO+ violet); thallus usually K+ orange to red (orange, needle-shaped crystals), rarely K+ yellow or K– (no crystals); with norstictic and/or stictic acid ................................................................................................................. B. nashii Outermost exciple cells without aeruginose pigment (HNO3–); thallus K– or K+ yellow (no crystals); with 2'-O-methylperlatolic acid ......................... B. dispersa 21(17) Exciple deep brown, without aeruginose pigment (HNO3–) .............................. 22 424 Exciple and epihymenium olive brown or bluish green, rarely fuscous brown, always with an aeruginose pigment (HNO3+ violet) .......................................... 29 22(21) Spores septum distinctly thickened during some stages of the ascospore ontogeny ............................................................................................................................ 23 Spore septum not thickened during the entire ascospore ontogeny ................... 25 23(22) Apothecial disc deep brown, blackening with age; young apothecia lecanorine, becoming lecideine with age (i.e. thalline margin darkening and excluded by expansion of proper exciple), medulla amyloid (I+ blue, test carefully on thin thalli!) .......................................................... B. mamillana (Tuck.) W. A. Weber Apothecial disc deep black, not brown; young apothecia lecideine; medulla not amyloid (I–) ....................................................................................................... 24 24(23) Thallus thin, ± continuous, pale yellowish, ivory or yellowish green, containing xanthones; all spot test reactions negative; apothecial margin thin, indistinct; exciple narrow, reduced to a few leptodermatous hyphae similar in structure and orientation to the paraphyses (aethalea-type); outer exciple cells strongly inflated, carbonized by a brown pigment cap; spores smooth to microrugulate; septum thickening ± persistent; conidia filiform .......................................... B. prospersa Thallus moderately thickened, continuous, pale beige to pale brown; without xanthones, usually with atranorin and norstictic acid (K+ yellow to orange, forming crystals, P+ yellow), rarely also with gyrophoric acid (C+ pink, fleeting); apothecial margin thick, prominent; exciple broad, not reduced, of mesodermatous interwoven hyphae similar in structure to the hypothecium, outer 425 exciple cells barely inflated, brown pigmentation ± evenly distributed throughout (leptocline-type); spores microrugulate to rugulate; septum thickening soon reduced; conidia bacilliform to fusiform ......... B. subdisciformis (Leight.) Vain. 25(22) Hypothecium hyaline; apothecia remaining immersed between areoles, disc ± irregularly deformed by adjoining areoles; with lecanoric acid (C+ fleeting pink, KC+ fleeting pink or C–, KC–); subalpine to alpine, rare, known from San Francisco Peaks and White Mountains (Arizona), Lake Peak and Sierra Blanca Peak, (New Mexico) ............................................................... B. eganii Bungartz Hypothecium reddish brown; apothecia emergent; disc circular, not deformed; without lecanoric acid (C–, KC–); coastal or more widely distributed montane species ................................................................................................................ 26 26(25) Thallus pale gray to white, not beige or deep brown; spores distinctly rugulate; with divaricatic acid and biosynthetically related substances ................................ ................................................................................................... B. tesserata Körb. Thallus deep brown to olive brown, rarely pale; spores smooth or microrugulate; no divaricatic acid or related substances ............................................................ 27 27(26) Thallus distinctly areolate, thin to moderately thickened, deep brown to olive brown; with distinct black hypothallus delimiting the thallus and often extending between the areoles, inland ................................................................ B. tyrolensis Thallus rimose to rimose-areolate, thin, not olive, usually deep brown, rarely pale brown; without a hypothallus, coastal and inland ............................................. 28 426 28(27) Premature spores with evenly thickened wall, ± globose, becoming ellipsoid with age; margin prominent, thick, rarely excluded; outer exciple > 20 µm wide in cross section, conidia bacilliform .................................... B. christophii Bungartz Spores of all stages thin-walled, ellipsoid to oblong; lecideine margin thin, usually excluded; outer exciple < 20 µm in cross section; conidia filiform ........... ...................................................................................................... B. pullata Tuck. 29(21) Medulla (at least in parts) amyloid (I+ blue) ..................................................... 30 Medulla not amyloid (I–) ................................................................................... 34 30(29) Medulla K–, KC+ pink and C+ pink (gyrophoric acid), rare ............... B. uberior (Note: In the Sonoran region only parasitic material on Schaereria fuscocinerea has so far been observed. The species is keyed out here, because non-parasitic type material of B. malmei Lynge from Novaya Zemlya (arctic Russia) is anatomically, morphologically and chemically identical and possibly a rare form of B. uberior that develops independent thalli.) Medulla K+ orange to red (orange, needle-shaped crystals), KC– and C–; with norstictic acid ....................................................................................................... 31 31(30) Young spores with distinct septum thickening, medulla strongly amyloid, even if tested on thallus surface (I+ blackish blue), apothecia remaining immersed, not emerging from thallus, often deformed by adjoining areoles, rarely with a thalline veil; differentiated into distinct, hyaline subhymenium gradually merging into pale to deep reddish brown hypothecium ................... B. lacteoidea de Lesd. 427 Young spores without septum thickening, medulla amyloid, but reaction obscured if tested on thallus cortex (I+ blue), apothecia immersed to sessile, deformed or not, with or without thalline veil, hypothecium not differentiated from subhymenium, but sometimes hyaline ...................................................... 32 32(31) Apothecia soon sessile, with thick and prominent lecideine margin; rarely with thalline veil; inner excipular hyphae broad, radiating from deep reddish brown hypothecium, with a narrow, hyaline transition zone and distinctly swollen, pigmented outer exciple cells (dispersa-type); conidia > 5 µm long, ± fusiform .. ...................................................................................................... Buellia sheardii Apothecia immersed to adnate, with thin lecideine margin, young apothecia usually with distinct thalline veil; inner excipular hyphae thin, ± reduced and hyaline, not distinctly radiating from the hypothecium, similar in structure and orientation to the paraphyses, with moderately swollen, pigmented outer exciple cells (aethalea-type); conidia , 5 µm long, ± bacilliform .................................. 33 33(32) Thallus with atranorin and norstictic acid; apothecia initially immersed, becoming sessile, circular, not deformed, young apothecia frequently with thalline veil, hypothecium reddish brown throughout; montane, common ............ ......................................................................................... B. spuria (Schaer.) Anzi Thallus with norstictic acid only, no other substances; apothecia immersed, not becoming sessile, without thalline veil, often deformed (“comma-shaped”); hypothecium hyaline throughout (at least in all specimens from Sonoran Region); montane, rare ...................................................................................... B. aethalea 428 34(29) Apothecia remaining immersed, not emerging, often deformed (“commashaped”), rarely circular; all Sonoran specimens with hyaline to pale brown hypothecium; with norstictic acid only, K+ orange to red (orange, needle-shaped crystals); inland ................................................................................... B. aethalea Apothecia initially immersed, emerging, not deformed; hypothecium dark reddish brown; norstictic acid absent, K+ yellow (no crystals forming) or K–; coastal ................................................................................................................. 35 35(34) Thallus white to dark gray, not olive or brown; with atranorin and 2'-Omethylperlatolic acid; K+ yellow; young apothecia immersed, but not aspicillioid, becoming adnate to sessile, usually emerging with thalline veil ........ ................................................................................... B. stellulata (Taylor) Mudd Thallus brown or olive gray; no secondary metabolites detected; K–; young apothecia immersed, appearing aspicillioid (“bursting through the thallus surface”), becoming adnate to sessile with age, rarely with remains of necrotic material attached to margin (indistinct thalline veil) .......................................... 36 36(35) Thallus reddish brown (“leather-colored”), rimose, not delimited by distinct hypothallus, exciple deeply aeruginose, ascospores 10–15 x 6–9 µm; conidia 4–7 µm ........................................................................................... B. tergua Bungartz Thallus olive gray to brownish olive, distinctly areolate in the center, with intermediate undifferentiated transition zone along the margin, rising from distinct black, arachnoid hypothallus, exciple fuscous brown, ascospores 9–13 x 4–8 µm; conidia 2-5 µm .......................................................... B. ryanii Bungartz 429 ACKNOWLEDGEMENTS I am much indebted to my Ph.D. supervisor Dr. Thomas H. Nash III, who supported my research for the last five years and frequently offered much valuable advice for the research presented here. Other members of my graduate student committee have also been very supportive: Dr. Donald J. Pinkava, Dr. Robert W. Roberson, Dr. Bruce D. Ryan and Dr. John W. Sheard. I am grateful to Scott Bates, for reviewing the manuscript. Dr. John Sheard, University of Saskatchewan, Canada and Dr. Mayrhofer, University of Graz, Austria have kindly agreed to review the manuscript for Mycotaxon. Loans from the following herbaria to ASU are greatly appreciated: BM, CANL, COLO, FH, H, hb. Scheidegger, hb. Kalb, hb. Sheard, L, M, MICH, MSC, SBBG, UPS, W. Dr. John A. Elix, University of Canberra, Australia, analyzed selected specimens for me with HPLC. The study was supported by a Research Grant in Plant Systematics from the International Association of Plant Taxonomists (IAPT) and National Science Foundation Awards to ASU (DEB-0103738, DEB-9701111). A research visit to Michigan State University (MSU) in Fall 2003 was supported by a National Science Foundation Award (DBI0237401). The staff at MSU entered specimen data and my annotations into their database and made the data available for my research. LITERATURE CITED BIRKBECK, A. A., M. V. SARGENT & J. A. ELIX. 1990. The structure of the lichen depsidones fulgidin and isofulgidin. Australian Journal of Chemistry 43: 419-425. BUNGARTZ, F. & T. H. NASH III. 2004a. Buellia subalbula (Nyl.) Müll. Arg. and B. amabilis de Lesd., two species from North America with one-septate ascospores: 430 A comparison with Buellia ["Diplotomma"] venusta (Körb.) Lettau. pp. 49-66. In RAMBOLD, G. & P. DÖBBELER (eds.), Lichenological Contributions. Festschrift in honor of Hannes Hertel on occasion of his 65th birthday, Bibliotheca Lichenologica, 88. Gebrüder Borntraeger, Stuttgart. ———. 2004b. Buellia turgescens is synonymous with Buellia badia and must not be included in Amandinea. The Bryologist 107: 21-27. ———. 2004c. The Buellia aethalea-Group in the Greater Sonoran Desert Region with some reference to similar species known from North America. The Bryologist, submitted for publication February 2004. BUNGARTZ, F., C. SCHEIDEGGER & T. H. NASH III. 2002. Buellia dispersa A. Massal., a variable lichen species from semi-arid to arid environments of North America and Europe. pp. 19-35. In LLIMONA, X., H. T. LUMBSCH & S. OTT (eds.), Progress and Problems in Lichenology at the Turn of the Millennium, Bibliotheca Lichenologica, 82. J. Cramer, Berlin, Stuttgart. BUNGARTZ, F., T. H. NASH III & B. D. RYAN. 2004a. Morphology and anatomy of chasmolithic versus epilithic growth: a taxonomic revision of inconspicuous saxicolous Buellia species generally ascribed to the "Buellia punctata"-group from the Sonoran Desert Region. Canadian Journal of Botany, in press. BUNGARTZ, F., J. A. ELIX & T. H. NASH III. 2004b. The genus Buellia s.l. in the Greater Sonoran Desert Region: saxicolous species with one-septate ascospores containing xanthones. The Bryologist, submitted for publication February 2004. 431 BUSCHARDT, A. 1979. Zur Flechtenflora der inneralpinen Trockentäler unter besonderer Berücksichtigung des Vinschgaus. Bibliotheca Lichenologica. Band 10. J. Cramer, Vaduz. CULBERSON, C. F. & H. KRISTINSSON. 1970. A standardized method for the identification of lichen products. Journal of Chromatography 46: 85-93. CULBERSON, C. F. & A. JOHNSON. 1982. Substitution of methyl tert.-butyl ether for diethyl ether in standardized thin-layer chromatographic method for lichen products. 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The Annals and Magazine of Natural History (London) 4: 25-29. MARBACH, B. 2000. Corticole und lignicole Arten der Flechtengattung Buellia sensu lato in den Subtropen und Tropen. Bibliotheca Lichenologica 74: 1-384. MEYER, B. & C. PRINTZEN. 2000. Proposal for a standardized nomenclature and characterization of insoluble lichen pigments. Lichenologist 32: 571-583. 433 MIETZSCH, E., H. T. LUMBSCH & J. A. ELIX. 1994. WINTABOLITES (Mactabolites for Windows). Users manual and computer program. University Essen, Essen. MOBERG, R., A. NORDIN & C. SCHEIDEGGER. 1999. Proposal to change the listed type of the name Buellia, nom. cons. (Physciaceae, lichenized Ascomycota). Taxon 48: 143. NASH III, T. H., B. D. RYAN, C. GRIES & F. BUNGARTZ (eds.). 2002. Lichen Flora of the Greater Sonoran Desert Region 1: 1-532. NORDIN, A. 2000. Taxonomy and phylogeny of Buellia species with pluriseptate spores (Lecanorales, Ascomycotina). Symbolae Botanicae Upsalienses 33: 1-117. ORANGE, A., P. W. JAMES & F. J. WHITE. 2001. Microchemical Methods for the Identification of Lichens. British Lichen Society, London. PUSSWALD, W., G. KANTVILAS & H. MAYRHOFER. 1994. Hafellia dissa and H. levieri (lichenized Ascomycetes, Physciaceae), two corticolous and lignicolous species in Tasmania. Muelleria 8: 133-140. RICO, V. J., A. CALATAYUD & M. GIRALT. 2003. Buellia tesserata and Dimelaena radiata, two closely related species. Lichenologist 35: 117-124. SCHEIDEGGER, C. 1987. Buellia uberior und B. miriquidica (Physciaceae, Lecanorales), zwei lichenicole Krustenflechten auf Schaereria tenebrosa. Botanica Helvetica 97: 99-116. ———. 1991. Phytogeography of the genus Buellia (Physciaceae, Lecanorales) in Mediterranean Europe. Botanika Chronika 10: 211-220. 434 ———. 1993. A revision of European saxicolous species of the genus Buellia De Not. and formerly included genera. Lichenologist 25: 315-364. SHEARD, J. W. 1992. The lichenized ascomycete genus Hafellia in North America. The Bryologist 95: 79-87. SHEARD, J. W. & T. TØNSBERG. 1995. Rinodina stictica, a new corticolous, sorediate lichen species from the Pacific Northwest of North America. The Bryologist 98: 41-44. STEINER, J. 1907. Über Buellia saxorum und verwandte Flechtenarten. Verhandlungen der Zoologisch-Botanischen Gesellschaft in Wien\Verh. Zool.-Bot. Gesell. Wien 57: 340-371. THOMSON, J. W. 1997. American Arctic Lichens. 2. The Microlichens. The University of Wisconsin Press, Madison. VOBIS, G. 1980. Bau und Entwicklung der Flechten-Pycnidien und ihrer Conidien. Bibliotheca Lichenologica 14: 1-141. VOBIS, G. & D. L. HAWKSWORTH. 1981. Conidial lichen-forming fungi, pp. 245-273. In COLE, G. T. & B. KENDRICK (eds.), 1: Biology of Conidial Fungi. Academic Press, New York. WEBER, W. A. 1971. Four new species of Buellia (Lichenes) from western North and South America. The Bryologist 74: 185-191. ———. 1986. The lichen flora of the Galapagos Islands, Ecuador. Mycotaxon 27: 451497. 435 WHITE, F. J. & P. W. JAMES. 1985. New guide to microchemical techniques for the identification of lichen substances. British Lichen Society Bulletin 57: 1-41. ZAHLBRUCKNER, A. 1931. Catalogus Lichenum Universalis. Vol. 7. Borntraeger, Leipzig. 436 OTHER SPECIES DESCRIPTIONS: Buellia paniformis, B. subdisciformis, B. tesserata and B. uberior Descriptions of four species are provided here. These species were reported in Bungartz (2004, Publication 8), but not described in detail. BUELLIA PANIFORMIS W. A. Weber, Bryologist 74: 188. 1971. TYPE: MEXICO. BAJA CALIFORNIA. Guadalupe Island. 28°53'-29°10'N, 188°17'W, Lobster Camp, on basalt stones in canyon 1 mile west of the landing, 0-100 m, 19-29 April 1963, Weber & McCoy L-36577 (COLO–holotype!). Taxonomic note.—The species name derived from the Latin “panis”, meaning “bread”. It refers to the “bread-loaf” appearance of the areoles and is not to be confused with “panniformis” meaning with a “felt-like appearance”. Thallus (Fig. 53A,B) crustose, thick, dispersed, epilithic; bullate (forming “breadloaves”); prothallus absent; thallus surface shiny and smooth, not matt, pale brown to tawny yellow (“isabelline”), epruinose, rarely faintly pruinose, phenocorticate (with a very thick and distinctly pigmented phenocortex and distinct epinecral layer); lacking calcium oxalate crystals in the thallus medulla (H2SO4–, not forming clusters of needleshaped crystals). Apothecia lecideine; (0.3–)0.6–1.1(–1.2) mm in diameter; sessile; proper margin black, thin, rarely persistent, usually excluded with age; disc black, epruinose, plane, becoming strongly convex with age; proper exciple of the dispersa-type (Fig. 53C) sensu Scheidegger (1993), i.e. inner excipular hyphae distinct, not reduced, pigmented, prosoplectenchymatous (textura oblita), extending from the deep reddish 437 brown hypothecium (leptoclinoides-brown, textura intricata), outer excipular hyphae short-celled, cells angular, distinctly swollen (textura angularis) and ± carbonized with various amounts of a brown pigment (cf. elachista-brown, HNO3–), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachistabrown). Asci 8-spored, clavate, Bacidia-type. Ascospores (Fig. 53E-G) narrowly oblong to ellipsoid, usually not constricted, with obtuse ends, not curved, (11.0–)12.0–[13.6]– 15.2(–19.0) x (6.0–)6.0–[6.7]–7.5(–9.0) µm (n = 60); one-septate, proper septum becomes thickened early but only briefly during spore ontogeny, (± Physconia-type; Fig. 53E); ornamentation microrugulate; septum with septal pore canal, simple pore and undifferentiated pore plug; spore wall (Fig. 53G) differentiated into cracked, thin perispore (0.05–0.11 µm), narrow intermediate layer (< 0.04 µm), moderately thickened proper spore wall (0.27–0.38 µm) and thick endospore (0.37–0.46 µm). Pycnidia infrequent, urceolate to globose, unilocular, at maturity lined with short moderately branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore-type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicariatype (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 5.0–9.0 x 0.5–1.0 µm (n = 20). Chemistry.—Atranorin and chloroatranorin (biosynthetically related depsides). Diploicin, 3-demethylscensidin and isofulgidin (biosynthetically related depsidones). Thallus K+ yellow, P+ yellow, KC–, CK–. UV+ bright yellow. The thallus is not 438 amyloid, apothecia amyloid in Lugol’s (always test with concentrated Lugol's iodine or in the compound microscope; the reactions can be very weak!). → FIGURE 53. Buellia paniformis (Weber L-36577, COLO-holotype).—A. Thallus aspect of several distinctly bullate areoles.—B. Close-up of several bullate areoles and convex, lecideine apothecia.—C. Light micrograph of the dispersa-type exciple.—D. Light micrograph of the epihymenium (eh) covered by a thick epinecral layer (en).—E. Light micrograph of premature ascospores with distinct septal thickening (arrow).—F. Transmission electron micrograph of the ascospore: (as) ascus wall, (ap) ascoplasm.—G. Transmission electron micrograph of the ascospore wall: (s) mucilaginous sheath, (1) perispore, (2) intermediate layer, (3) proper wall, (4) endospore. 439 440 Substrate and ecology.—On volcanic rock (HCl–). Distribution (Fig. 9).—Known only from the type locality on Guadalupe Island and few localities along the foggy western shore of the Baja California peninsula. Additional specimens examined.—MEXICO. BAJA CALIFORNIA. Guadalupe Island. Weber L-36511, L-36576 (COLO); Nash 38445 (ASU). El Rosario near Al Aguajito. Mayrhofer 10282 (GZU). BUELLIA SUBDISCIFORMIS (Leight.) Vain., Etud. Lich. Brésil. 7: 167. 1890. Lecidea subdisciformis Leight., Lich. Fl. Great Brit.: 308. 1871. TYPE: UNITED KINGDOM. WALES. Caernarvonshire. Conway Mountain, 1851, Leighton s.n. (BM–holotype). For synonyms see Scheidegger (1993). Thallus (Fig. 54A,B) crustose, thin to moderately thickened, ± continuous, epilithic; rimose to rimose-areolate; prothallus absent or delimiting the thallus margin as a black outline; thallus surface smooth, usually matt or ± rarely shiny, usually ivory to pale brown, rarely grayish brown, epruinose, phenocorticate; lacking calcium oxalate crystals in the thallus medulla (H2SO4–, not forming clusters of needle-shaped crystals). Apothecia lecideine; (0.2–)0.4–0.6(–0.8) mm in diameter; soon adnate to sessile; proper margin black, prominent, usually persistent, rarely excluded with age; disc black, epruinose, plane, rarely becoming convex with age; proper exciple distinct, of the leptocline-type (Fig. 54C) sensu Scheidegger (1993), i.e. exciple thick and not distinctly differentiated into an inner and outer part, hyphae thin-walled (mesodermatous), 441 prosoplectenchymatous and usually ± densely interwoven (textura intricata), dull fuscous brown throughout, becoming ± carbonized by various amounts of a brown pigment (cf. elachista-brown, HNO3–), pigmentation continuous with the epihymenium, hypothecium deep reddish brown (leptoclinoides-brown, textura intricata); hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown). Asci 8-spored, clavate, Bacidia-type. Ascospores (Fig. 54D) oblong to ellipsoid, usually not constricted, with obtuse ends, not curved, (9.0–)11.3–[12.8]–14.2(–16.0) x (5.0–)5.8–[6.7]–7.6(–8.0) µm (n = 60); one-septate, proper septum becomes distinctly thickened during spore ontogeny (Physconia-type); ornamentation microrugulate to rugulate. Pycnidia rare, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicariatype (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform to fusiform, 7.0–14.5x 1.0–1.5 µm (n = 20). Chemistry.—Depsides: atranorin and gyrophoric acid. Depsidone: norstictic acid. Anthraquinones: secalonic acid C. Ergochromes (chemical structure of these not yet determined): eumitrin x, x2, y. K+ yellow, P+ yellow, C–, rarely C+ pink, KC–, rarely KC+ pink, CK–. UV– (pale). The thallus is not amyloid, apothecia amyloid in Lugol’s (always test with concentrated Lugol's iodine or in the compound microscope; the reactions can be very weak!). Substrate and ecology.—On exposed siliceous rocks (HCl–). 442 → FIGURE 54. Buellia subdisciformis (Nash 12242, ASU).—A. Rimose thallus.—B. Close-up of the rimose areolate thallus, several apothecia have distinct lecideine margin.—C. Light micrograph of the leptocline-type exciple.—D. Overmature ascospores with distinct microrugulate ornamentation; the septal thickening is no longer visible at this stage. 343 444 Distribution (Fig. 11).—According to Scheidegger (1993) widespread throughout the Mediterranean Region. Apparently quite rare in the Sonoran Region, where only few specimens have been collected. Notes.—Scheidegger (1993) distinguishes the following morphologically, anatomically and ecologically similar taxa: Buellia subdisciformis (atranorin, norstictic and connorstictic acid, medulla I-), B. sardiniensis (atranorin, gyrophoric, norstictic and connorstictic acids, medulla I+ blue) and B. saxorum (atranorin, gyrophoric and lecanoric acids, medulla I+ blue). Two specimen examined from Europe [Great Britain: Henssen 18818a and Italy: Nash 24836] have a non-amyloid medulla but contain both gyrophoric and norstictic acid. All Sonoran specimens also have a I– medulla but regularly contain atranorin and both norstictic and gyrophoric acid. Based on the negative iodine reaction of the medulla, this material as well as the specimens from Italy and Great Britain can be assigned to B. subdisciformis. It seems very likely that B. sardiniensis should be treated as a synonym of B. subdisciformis. Scheidegger (1993) suggested that B. sardiniensis could possibly be treated as a chemical strain of B. saxorum. In contrast to the iodine negative material from the Sonoran Region, material from Scheidegger’s personal collection clearly belongs to B. saxorum because of the iodine deep blue medullary reaction. The material also does not contain norstictic or connorstictic acid. It is possible that all three taxa belong to the same species and the name B. saxorum would then have taxonomic priority. Without additional specimens available from the Sonoran Region this conclusion seems, however, premature. 445 Selected specimens examined.—FRANCE. PYRÉNÉES-ORIENTALES. Scheidegger Inv. Nr. 7619, 8376 (hb. Scheidegger). ITALY. SARDINIA. Nash 24836 (ASU); Scheidegger Inv. Nr. 10782 (hb. Scheidegger). MEXICO. SINALOA. Nash 12242, 12250 (ASU). NAYARIT. Nash 39191 (ASU). UNITED KINGDOM. CHANNEL ISLANDS. Henssen 18818 (ASU). BUELLIA TESSERATA Körb., Parerg. Lich.: 189. 1860. TYPE: NORWAY. On shale, Hübner & Kurr s.n. [original label data: “An Schieferfelsen Norwegens von Hübner und Kurr gesammelt”] (L–lectotype selected by Foucard et al. 2002, UPS–isolectotype). Buellia fimbriata (Tuck.) Sheard, Bryologist 72: 221. 1969.—Rinodina radiata var. fimbriata Tuck., Synops. No. Amer. Lich. 1: 205. 1882. TYPE: U.S.A. CALIFORNIA. San Francisco Co. Cliffs at the Mission Dolores. On rocks on the coast, Bolander s.n. (FHTUCK–lectotype selected by Sheard 1969). For additional synonyms see Rico et al. (2003) and Scheidegger (1993). Taxonomic note.—Rico et al. (2003) synonymized B. fimbriata with B. tesserata. This name change is problematic and is followed here only reluctantly. According to Scheidegger (1993) the type of B. tesserata is only known from the type locality. The type specimen must have been collected before the species was described in 1860. Because then no additional specimens have been found in Norway (Foucard et al. 2002), it is very unlikely that this common Mediterranean species actually grows in Norway and the type material may have been labeled incorrectly. A more elegant solution would have 446 been to conserve the name B. fimbriata against B. tesserata because B. tesserata is based on a dubious type collection. Thallus (Fig. 55A,B) crustose, thin to moderately thickened, ± continuous, epilithic; areolate; prothallus usually distinct, black, fimbriate along the thallus outline; thallus surface matt and smooth, usually white, rarely pale gray, often pruinose, phenocorticate; lacking calcium oxalate crystals in the thallus medulla (H2SO4–, not forming clusters of needle-shaped crystals). Apothecia lecideine; (0.2–)0.3–0.4(–0.5) mm in diameter; immersed, rarely emergent; proper margin black (or color masked by grayish remains of necrotic thalline material, thalline veil), thin, rarely persistent, usually excluded with age; disc black, often whitish pruinose, plane, rarely becoming convex with age; proper exciple narrow, poorly differentiated, of the aethalea-type (Fig. 55C) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), often reduced, similar in structure and orientation to the paraphyses, transient with the deep reddish brown hypothecium (leptoclinoides-brown, textura intricata), outer excipular hyphae parallel, moderately swollen (textura oblita) and usually strongly carbonized with various amounts of a brown pigment (cf. elachista-brown HNO3–), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown). Asci 8-spored, clavate, Bacidia-type. Ascospores (Fig. 55D-F) oblong to ellipsoid, usually not constricted, with obtuse ends, not curved, (9.0–)9.0– [9.8]–10.6(–12.0) x (4.0–)4.6–[5.2]–5.7(–6.5) µm (n = 60); one-septate, proper septum narrow, not thickening during spore ontogeny (Beltraminea (=Buellia)-type); 447 ornamentation distinctly rugulate; septum with septal pore canal, simple pore and undifferentiated pore plug; spore wall (Fig. 55G) differentiated into fractured, thin perispore (0.16–0.26 µm), narrow intermediate layer (< 0.03 µm), moderately thickened proper spore wall (0.18–0.25 µm) and moderately thickened endospore (0.17–0.25 µm). Pycnidia infrequent, urceolate to globose, unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 6.0–11.0 x 1.0–1.5 µm (n = 20). Chemistry.—Depsides: atranorin and chloroatranorin (biosynthetically related); haematommic acid and methyl-β-orsellinate (artifacts from hydrolysis of atranorin); divaricatic, nordivaricatic, 3-chlorodivaricatic acid (biosynthetically related); 2'-Omethylperlatolic acid; 3,5-dichloro-2'-O-methylanziaic acid; 3,5-dichloro-2'-Omethylnorimbricaric acid (new lichen substance, not previously reported in nature, Elix, pers. comm.). Rarely also with the following depsidones: norstictic, cryptostictic and stictic acid (biosynthetically related) and a trace of an unknown substance. Usually K+ yellow to red, P– or + faintly yellow, C–, rarely C+ fleeting pink, KC–, rarely KC+ fleeting pink, CK– (reactions often weak). UV– (pale). The thallus is not amyloid, apothecia amyloid in Lugol’s (always test with concentrated Lugol's iodine or in the compound microscope; the reactions can be very weak!). Substrate and ecology.—A distinctly maritime species, typically found on siliceous rocks (HCL–) close to the seashore. 448 → FIGURE 55. Buellia tesserata (A-E. light micrographs of Nash 29731, ASU, F.G. Transmission electron micrograph of Bungartz 3116, hb. Bungartz).—A. Areolate thallus aspect.—B. Close-up of the fimbriate prothallus (arrow).—C. The aethalea-type exciple.—D. Mature ascospores with distinct rugulate ornamentation.—E. Mature ascospores with thin median septum.—F. Mature ascospore.—G. Ascospore wall: (s) mucilaginous sheath, (1) perispore, (2) intermediate layer, (3) proper wall, (4) endospore. 449 450 Distribution (Fig. 11).—In the Sonoran Region common on coastal rock along the shores of southern California and the Baja California peninsula; also known from coasts of the Mediterranean (Turkey, Greece, Italy, Portugal, France, Morocco and Algeria) and the Cape Verde Islands (see Rico et al. 2003). Notes.—The species is very closely related to Dimelaena radiata (Tuck.) Hale & W.L. Culb. Both species share an identical chemistry, anatomy and spore structure (Rico et al. 2003). The two taxa should consequently be united in the same genus. Emending the genus concept of Dimelaena to include taxa with a fimbriate margin is, however, problematic because a lobate margin is the major criterion currently used to segregate Dimelaena from Buellia. Although the two taxa are very similar, they nevertheless maintain a distinctly different thallus morphology when growing together in the same habitat. This suggests that the different morphology is not an environmental variation, but that the two taxa are indeed separate species. Selected specimens examined.—FRANCE. PYRÉNÉES-ORIENTALES. Scheidegger Inv. Nr. 7541, 8373 (hb. Scheidegger). GREECE. Sipman 42863 (B). ITALY. SARDINIA. Scheidegger Inv. Nr. 10253 (hb. Scheidegger); Nash 25097 (ASU). MEXICO. BAJA CALIFORNIA. Scheidegger 40-59 (hb. Scheidegger); Wetmore 70263, 72260 (MIN); Kalb 24673a, 24750a (hb. Kalb), Weber L-43032 (COLO); Nash 4575b, 26188 (ASU). MEXICO, BAJA CALIFORNIA SUR. Wetmore 70225, 72167 (MIN); Moberg 10377 (UPS); Marsh 6078, Nash 33893b, 16929 (ASU). SONORA. Nash 10986b (ASU). SPAIN. ANDALUCÍA. Almería. Llimona s.n. (Vězda Lich. Sel. Exs.1325, ASU). USA. California. Los Angeles Co. Hasse 9b (ASU); Sheard 5015 (CANL); Nash 32114 (ASU); Weber L- 451 42165 (COLO). Marin Co. Ryan 21929 (ASU); Weber L-60792 (COLO). San Diego Co. Bratt 5622 (SBBG). San Luis Obispo Co. Riefner 88-40 (ASU). San Mateo Co. Ryan 21975a (ASU). Santa Barbara Co. Bratt 3665 (SBBG); Wetmore 73801 (MIN); Sheard 5094b (CANL); Nash 32749 (ASU); Weber L-81086 (COLO). Ventura Co. Bratt 7846 (SBBG); Marsh 7832, Nash 38741 (ASU). BUELLIA UBERIOR Anzi, Atti. Soc. Sci. Nat. 9: 252. 1866. Holotype: ITALY. Lombardia, Alpe Braulio, valle Zebru, Anzi s.n. (M-lectotype, HNYL-9321-isotype). For additional synonyms see Scheidegger (1987). Thallus (Fig. 56A,C) crustose, moderately thickened, ± continuous, epilithic; areolate; prothallus absent; thallus surface matt and dull, not shiny, usually deep gray, rarely bluish gray, epruinose, phenocorticate; lacking calcium oxalate crystals in the thallus medulla (H2SO4–, not forming clusters of needle-shaped crystals). Apothecia lecideine; (0.1–)0.2– 0.3(–0.4) mm in diameter; remaining immersed; proper margin black, thin, rarely persistent, usually excluded with age; disc black, epruinose, plane, usually becoming slightly convex with age; proper exciple narrow, poorly differentiated, of the aethaleatype (Fig. 56B) sensu Scheidegger (1993), i.e. inner excipular hyphae narrow, hyaline, prosoplectenchymatous (textura oblita), often reduced, similar in structure and orientation to the paraphyses, transient with the deep reddish brown hypothecium (leptoclinoides-brown, textura intricata), outer excipular hyphae parallel, moderately swollen (textura oblita) and usually strongly carbonized with various amounts of brown 452 and aeruginose pigments (cf. elachista-brown & cinereorufa-green, HNO3+ violet), pigmentation continuous with the epihymenium; hymenium hyaline, not inspersed; paraphyses simple to moderately branched, apically swollen, with a brown pigment cap (cf. elachista-brown) and a diffuse aeruginose pigment (HNO3+ violet, cinereorufagreen). Asci 8-spored, clavate, Bacidia-type. Ascospores (Fig. 56D) broadly ellipsoid, rarely ± constricted, with obtuse ends, not curved, (8.0–)9.1–[10.1]–11.1(–14.0) x (4.5–)4.9–[5.6]–6.3(–7.0) µm (n = 60); one-septate, proper septum narrow, not thickening → FIGURE 56. Buellia uberior (Nash 25408, ASU).—A. Areolate thallus forming an insular patch in the areolate thallus of Schaereria fuscocinerea.—B. Light micrograph of the aethalea-type exciple.—C. Close-up of the parasitic thallus.—D. Mature, slightly constricted ascospores. 453 454 during spore ontogeny (Beltraminea (=Buellia)-type); ornamentation striate (visible in DIC, but often inconspicuous). Pycnidia unilocular, at maturity almost entirely occupied by densely branched conidiophores; conidiogeneous cells mostly terminal, rarely also intercalary (cf. conidiophore type V sensu Vobis 1980); pycnidial ontogeny similar to the Umbilicaria-type (sensu Vobis 1980 and Vobis & Hawksworth 1981); conidia simple, bacilliform, 4.5–7.0 x 0.5–1.0 µm (n = 20). Chemistry.—Typically with the depside gyrophoric acid; Scheidegger (1993) also reports stictic acid from some of the samples he examined. K–, P–, C+ pink, KC+ pink (test the white thallus medulla). UV– (dark). The thallus is amyloid, apothecia amyloid in Lugol’s (always test with concentrated Lugol's iodine or in the compound microscope; the reactions can be very weak!). Substrate and ecology.—The only known specimens from the Sonoran Desert Region form distinct parasitic patches in Schaereria fuscocinerea (Nyl.) Clauzade & Cl. Roux. Distribution (Fig. 11).—Closely associated with its host and rarely collected, e.g. along the Mogollon Rim (Arizona). Notes.—In the Sonoran Region only parasitic material on Schaereria fuscocinerea has so far been observed. Non-parasitic type material of B. malmei from Novaya Zemlya is anatomically, morphologically and chemically identical and possibly a rare form of B. uberior that develops independent thalli. 455 TAXONOMIC PERSPECTIVES Taxonomy must not be based on arbitrarily selected characters or personal preferences but on scientific evidence. The Linnaean nomenclature provides a well established framework for taxonomy, but has been criticized as too rigid and artificial (http://www.ohiou.edu/phylocode/). This criticism must not be ignored! Linnaean nomenclature will not survive if new genera are based on very little evidence. Ideally, biological nomenclature should be in agreement with evolutionary theory and thus reflect the phylogeny of a species. Species must be recognized as evolutionary and ecological units and not as static concepts. Hierarchical ranks like genus or family should reflect how species have evolved and how they are related. However, nomenclature also serves a second purpose. Biological sciences rely on a common framework of naming organisms. The Linnaean type concept ensures that species concepts are not purely theoretical, but based on empirical reference points. Even though species are evolutionary and ecological units, they remain abstract without reference collections and diagnostic characters. A ranking, hierarchical system is not arbitrary, if it provides a convenient framework for the biological sciences. Thus, for example, most botanists know what members of the “sunflower family” look like. It may be more difficult to convey the concept of a lichen genus, but taxa will ultimately only be accepted if they can be recognized. The challenge of taxonomy is not to distinguish every difference but to evaluate which characters are meaningful. Nimis (1998) suggested five criteria to test whether new generic segregates should be accepted, paraphrased as follows: 456 (1) a genus should be based not only on classical but also on molecular evidence; this is not yet the case for most segregates of Buellia s.l. (2) a genus should be tested with a phylogenetic analysis; first attempts to analyze Buellia s.l. have been presented but are by no means exhaustive (Imshaug 1951; Scheidegger 1993; Marbach 2000; Nordin 2000). (3) a genus should be based on several discriminatory characters, not on a single character alone; this is not the case for genera like Amandinea. (4) the majority of species in a genus should be taken into account before a new generic segregate is accepted; so far only subsets of Buellia s.l. have been examined and the classical data are insufficient to examine large data sets including representative members of all generic segregates. (5) a new genus should at least convey new information; some segregates of Buellia like Hafellia or the Buellia alboatra-group could be accepted because they seem to be fairly well delimited, most other genera, however, convey very little information. Even the most sophisticated taxonomies reflect hypotheses and are thus subjective. These hypotheses change with our understanding of the natural world. Striving for a balance between a working taxonomy and an ideal phylogeny will thus remain a challenge. 457 REFERENCES ABU-ZINADA, A. H., D. L. HAWKSWORTH & H. A. BOKHARY. 1986. The lichens of Saudi Arabia, with a key to the species reported. 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New guide to microchemical techniques for the identification of lichen substances. British Lichen Society Bulletin 57: 1-41. WIRTH, V. 1994. Checkliste der Flechten und flechtenbewohnenden Pilze Deutschlands eine Arbeitshilfe [Checklist of the lichens and lichenicolous fungi of Germany]. Stuttgarter Beitrage zur Naturkunde, Serie A 517: 1-63. ———. 1995. Die Flechten Baden-Württembergs, Teil 1 & 2. Ulmer, Stuttgart. ZAHLBRUCKNER, A. 1898. Beiträge zur Flechtenflora Niederösterreichs V. Verhandlungen der Zoologisch-Botanischen Gesellschaft in Wien 48: 349-370. ———. 1926. Lichenes (Flechten). B. Spezieller Teil: Schlauchflechten (Ascolichenes); Hymenolichenes (Basidiomycetenflechten). Nachträge zu Teil 1, Abteilung 1 bis Ende 1906. Zweite Auflage. 8, pp. 61-236. In ENGLER, A. & K. PRANTL (eds.), Die natürlichen Pflanzenfamilien nebst ihren Gattungen und wichtigeren Arten. Borntraeger, Leipzig. 479 ———. 1931. Catalogus Lichenum Universalis. Vol. 7. Borntraeger, Leipzig. APPENDIX I LIST OF SPECIMENS EXAMINED WITH HPLC Specimens examined with HPLC by Dr. J. A. Elix (Australian National University, Canberra), artifacts from the hydrolysis of atranorin are marked by (*). B. christophii Weber L-42662 (COLO): parietin (not detected in any specimens by TLC and most likely a contamination from surrounding Caloplaca sp.). B. concinna Nash 11287 (ASU): arthothelin, 4,5-dichloro-6-O-methylnorlichexanthone, 6-Omethylarthothelin. Nash 18578 (ASU): 4,5-dichloronorlichexanthone, arthothelin, thiophanic acid. Nash 22836 (ASU): 4,5-dichloro-6-O-methylnorlichexanthone, 6-Omethylarthothelin. Nash 38098 (ASU): 4,5-dichloronorlichexanthone, arthothelin, isoarthothelin, thiophanic acid. Nordin 5178 (ASU): 4,5-dichloronorlichexanthone, arthothelin, isoarthothelin, thiophanic acid. Ryan 24231 (ASU): gyrophoric acid, 6-O-methylarthothelin. Ryan 24490 (ASU): 6-O-methylarthothelin Ryan 26137b (ASU): arthothelin, asemone, thiophanic acid. Scheidegger 7840 (hb. Scheidegger): arthothelin. Thiers 34410 (ASU): orsellinic acid, lecanoric acid, gyrophoric acid, 6-Omethylarthothelin B. eganii Nash 36934 (ASU): lecanoric acid, 5-O-methylhiascic acid. Nash 11720 (ASU): lecanoric acid, 5-O-methylhiascic acid. B. halonia Nash 17166 (ASU): 2,5-dichloronorlichexanthone, 4,5-dichloronorlichexanthone, 2,4dichloronorlichexanthone, arthothelin, atranorin, thiophanic acid. Nash 38387 (ASU): norstictic acid, eumitrin x & y (chemical structure of these ergochromes not yet determined!), isoarthothelin, thiophanic acid. Nash 40017 (ASU): isoarthothelin, thiophaninic acid, thiophanic acid. Nash 8027b (ASU): 2,5-dichloronorlichexanthone, 4,5-dichloronorlichexanthone, 2,4dichloronorlichexanthone, arthothelin, thiophanic acid. Weber L-42632 (COLO): 2,5-dichloronorlichexanthone, isoarthothelin, thiophanic acid. Wirth 22809 (STU): 2,5-dichloronorlichexanthone, 2,7-dichloronorlichexanthone, 5,7dichloronorlichexanthone, isoarthothelin, atranorin, thiophanic acid. B. lacteoidea Nash 22588 (ASU): connorstictic acid, norstictic acid, gyrophoric acid, atranorin. Nebecker 2166 (ASU): connorstictic acid, methyl-β-orsellinate*, norstictic acid, haematommic acid*, atranorin. B. lepidastroidea Wetmore 63852 (ASU): gangaleoidin, fulgidin, 3-dechlorodiploicin, isofulgidin, atranorin, chloroatranorin, diploicin. Weber L-42606 (ASU): methyl-β-orsellinate*, haematommic acid*, fulgidin, 3dechlorodiploicin, isofulgidin, atranorin, diploicin, chloroatranorin. 481 B. mamillana Marsh 5918 (ASU): cryptostictic acid, stictic acid, norstictic acid, ?unknown trace, divaricatic acid, atranorin, 2'-O-methylperlatolic acid, 4,5-dichlorolichexanthone. Nash 29793 (ASU): cryptostictic acid, stictic acid, methyl-β-orsellinate*, norstictic acid, methylstictic acid, hypostictic acid, haematommic acid*, atranorin, 4,5dichloro-3-O-methylnorlichexanthone, 4,5-dichlorolichexanthone. Nash 37836 (ASU): cryptostictic acid, stictic acid, norstictic acid, methylstictic acid, hypostictic acid, atranorin, 4,5-dichloro-3-O-methylnorlichexanthone, 4,5dichlorolichexanthone. Nash 7984 (ASU): cryptostictic acid, stictic acid, norstictic acid, methylstictic acid, hypostictic acid, atranorin, 4,5-dichlorolichexanthone. Weber S-8881 (COLO): norstictic acid, 4,5-dichloro-3-O-methylnorlichexanthone, 4chlorolichexanthone, 4,5-dichlorolichexanthone. B. navajoensis Nash 30659c (ASU): 4,5-dichloronorlichexanthone, arthothelin, atranorin, thiophanic acid. B. oidalea Ryan 31282 (ASU): orsellinic acid, gyrophoric acid, atranorin, 2'-O-methylperlatolic acid. B. paniformis Bungartz 3161 (hb. Bungartz): methyl-β-orsellinate*, haematommic acid*, isofulgidin, atranorin, chloroatranorin, diploicin. Nash 38445 (ASU): 3-demethylscensidin, isofulgidin, atranorin, chloroatranorin, diploicin. Weber L-36511 (COLO): isofulgidin, atranorin, chloroatranorin, diploicin. B. pullata: Nash 41227 (ASU): no substances detected! B. regineae Nash 26092 (ASU): fulgidin, 3-dechlorodiploicin, isofulgidin, atranorin, chloroatranorin, diploicin, brialmontin 1. Nash 34319 (ASU): dechlorogangaleoidin, gangaleoidin, norgangaleoidin, virensic acid, chlorolecidioidin, fulgidin, 3-dechlorodiploicin, isofulgidin, atranorin, chloroatranorin, diploicin. Nash 38303b (ASU): methyl-β-orsellinate*, haematommic acid *, chlorohaematommic acid*, fulgidin, isofulgidin, atranorin, chloroatranorin, diploicin. B. ryanii Wetmore 74077 (ASU): no substances detected! B. sequax Nash 8035 (ASU): no substances detected! B. spuria Weber Exs. 98 (COLO): fulgidin, 3-dechlorodiploicin, isofulgidin, atranorin, chloroatranorin, diploicin. Weber L-65225 (COLO): methyl-β-orsellinate, norstictic, atranorin, chloroatranorin. 482 Wetmore 18467 (MIN): cryptostictic acid, stictic acid, methyl-β-orsellinate*, norstictic acid, methylstictic acid, hypostictic acid, haematommic acid*, atranorin, chloroatranorin. B. subaethalea Nash 25353 (ASU): cryptostictic acid, norstictic acid, methylstictic acid, 4,5dichlorolichexanthone. Weber L-36928 (COLO): norstictic acid, 4,5-dichlorolichexanthone. B. subdisciformis Nash 24836 (ASU): norstictic acid, eumitrin x, x2, y (chemical structure of these ergochromes not yet determined!), secalonic acid c, gyrophoric acid, atranorin. B. tesserata Marsh 6084 (ASU): ?unknown trace, 3-chlorodivaricatic acid, atranorin. Nash 29731d (ASU): 3,5-dichloro-2'-O-methylnorimbricaric acid (new lichen substance, not previously reported in nature!), 3,5-dichloro-2'-O-methylanziaic acid. Nash 37838 (ASU): cryptostictic acid, stictic acid, methyl-β-orsellinate*, norstictic acid, haematommic acid*, atranorin, chloroatranorin. Weber L-42165 (COLO): divaricatic acid, atranorin, 2'-O-methylperlatolic acid. Weber L-81089 (COLO): 3-chlorodivaricatic acid. B. trachyspora Weber L-65279 (COLO): 4,5-dichloro-3-O-methylnorlichexanthone, 4,5dichlorolichexanthone. B. triphragmoides Nash 38513a (ASU): no substances detected. B. tyrolensis Ryan 10978 (ASU): 2'-O-methylperlatolic acid. 483 APPENDIX II CHARACTER CODING FOR THE PHYLOGENETIC ANALYSIS Character Coding (1) Substrate: 0. on bark, 1. on rock, 2. on soil, 3. both on rock and bark. Note: Marbach (2000) suggested that corticolous species were not closely related to saxicolous taxa. To evaluate this hypothesis information about the substrate was included even though substrate preference must be regarded as an ecological rather that taxonomic character. (2) Prothallus: 0. absent, 1. present. Note: It is problematic to include variable thallus characters in any analysis. Nevertheless most of species can be distinguished by the presence or absence of a prothallus. Species like B. spuria, B. tyrolensis or B. stellulata usually have a distinct black prothallus. The extent to which this prothallus is developed may be substrate dependant. Thus, on hard, impenetrable rock a prothallus frequently becomes more exuberant, extending between the areoles, thus forming a hypothallus. Other species nevertheless never form a prothallus even on hard, impenetrable substrates (e.g., B. dispersa, B. nashii or B. paniformis). (3) Conidia: 0. bacilliform (short & broad; most conidia < 5 µm), 1. fusiform (medium length and ± tapered; most conidia 5-14 µm), 2. filiform (long and usually curved; most conidia > 14 µm). Note: Nordin (2000) did not include condial length because species with pluriseptate spores show no or little variation. The character has been used to segregate Amandinea from Buellia. It was included here to test whether species with filiform conidia were supported as a distinct group. (4) Parasitism: 0. not parasitic (not associated with a host lichen), 1. facultative parasitic (distinct, separate thallus developing with age), 2. obligate parasite (thallus not distinctly separate from the host). Note: Some species of Buellia s.l. show parasitic tendencies or are even obligate parasites. The character was included because Marbach (2000) and Kalb (2004) suggested to transfer the facultative juvenile parasite B. badia into the separate genus Monerolechia. Like substrate preferences parasitism is strictly an ecological rather than a taxonomic character. (5) Thallus lobation: 0. squamules or lobes absent, 1. squamules or lobes present. Note: Nordin (2000) did not include this character suggesting that the formation of thallus lobes in placodioid taxa was an autapomorphy. Nordin & Mattsson (2001), however, included this vegetative character to compare crustose, foliose and fruticose taxa. The character is included here without deciding a priori whether it is informative or not. There is no distinct difference between subsquamulose, squamulose or even lobate thalli. All these thalli have areoles with tendencies to form some sort of thallus lobation. (6) Thallus continuity: 0. thallus initially forming a continuous crust, becoming rimose to rarely rimose areolate, 1. thallus forming distinct areoles, which grow aggregated or dispersed and may be bullate, granular, verrucose, subsquamulose or rarely squamulose. Note: Two general tendencies of thallus development in saxicolous species can be distinguished: Some thalli form initially as a continuous crust and secondarily become fissured (rimose) or cracked (rimose-areolate). Distinctly different from this development is a thallus developing from dispersed, separate granules which often become flattened areoles. Admittedly the two ontogenies are often difficult to distinguish. 485 (7) Iodine reaction of the medulla: 0. negative in all specimens, 1. positive at least in some of the specimens. Note: The reaction of the thallus medulla with concentrated Lugol’s iodine is diagnostic in most species. Only B. aethalea shows some variation (Scheidegger 1993). The species was coded as I+ because at least the Sonoran specimens all react I+ blue. (8) Soredia: 0. absent, 1. present. Note: Only very few species of Buellia s.l. form soredia and the character may be significant. (9) calcium oxalates presence: 0. absent, 1. present. Note: Species, which were not examined with X-ray diffraction were tested with 10% H2SO4 (Bungartz & Nash 2004a, Publication 4). (10) Apothecium diameter: 0. small (< 0.5 mm), 1. medium (0.5-1 mm), 2. large (> 1 mm). (11) Apothecium position: 0. soon sessile, 1. long or permanently remaining immersed. (12) Thalline exciple: 0. absent, 1. present. Note: A true, thalline exciple with a cortex and photobiont cells is present in the two Rinodina species and young apothecia of B. mamillana. All other species do not have a true thalline exciple. (13) Disc pruina: 0. absent, 1. present. (14) Pruina substance: 0. calcium oxalates, 2. xanthones. (15) Width of the proper exciple: 0. < 50 µm, 1. 50-100 µm, 2. > 100 µm. Note: This is another potentially problematic character because the width varies considerable among young and older apothecia. (16) Exciple type: 0. aethalea-type (rim distinctly darker than the inner exciple), 1. leptoclinoides-type (exciple dark throughout), 2. dispersa-type (middle part of young apothecia pale, eventually evenly pigmented in old apothecia), 3. Exciple pale throughout (Buellia mamillana was included here because the pigmentation of the excipular hyphae is initially very weak and only becomes darker in mature apothecia), 4. trachyspora-type (distinctly paraplectenchymatous inner exciple). 5. vilis-type (inner exciple hyaline and densely interwoven). Note: Nordin’s (2000) character no. 9 “Excipulum pigmentation” had to be modified. His pigmentation-types correspond to some extent to the exciple types described by Scheidegger (1993). Most exciple types sensu Nordin (2000) can therefore easily be assigned to the categories used here. However, Buellia vilis and B. trachyspora have very unique exciple types not found in any other species. Thus, two character states were added to the analysis. (17) Width of exciple cells: 0. > 4 µm, 1. < 4 µm. Note: This character may not be independent from the exciple type. (18) Hymenium inspersion: 0. oil droplets absent, 1. oil droplets present. (19) Ascus type: 0. amyloid reaction absent at the ascus tip (Lecanora-type), 1. amyloid reaction of the ascus flanks extends across the tip (Bacidia-type). (20) Hypothecium pigmentation: 0. hyaline in all specimens, 1. dark brown in all specimens, 2. both populations with pale or dark hymenium have been reported. Note: B. aethalea is the only species which has specimens either with a pale or dark brown hypothecium. Sonoran specimens all have a very weakly pigmented hypothecium. (21) Subhymenium pigmentation: 0. hyaline, 1. brown. Note: Almost all species of Buellia s.l. show no distinct differentiation of a subhymenium from the hypothecium. Both layers are usually deeply pigmented. However, in B. lacteoidea a distinct 486 differentiation can be observed. It would be misleading to code this species as polymorphic. The hypothecium of B. lacteoidea is clearly pigmented, but the subhymenium remains unpigmented for considerable time. (22) Spore size: 0. small (< 18 µm), 1. medium (18-20 µm), 2. large (20-28 µm), 3. very large (> 28 µm). (23) Perispore: 0. absent, 1. present. (24) Perispore thickness: 0. < 0.27 µm, 1. > 0.30 µm. (25) Proper wall thickness: 1. < 0.44 µm, 0. 0.48-0.74 µm, 2. > 0.82 µm. (26) Spore ornamentation: 0. absent, 1. present. (27) Apical wall thickenings: 0. absent, 1. present. (28) Lateral wall thickenings: 0. absent, 1. present. (29) Septal wall thickenings: 0. absent, 1. present. (30) Median septa: 0. one-septate, 1. additional median septa present. (31) Longitudinal septa: 0. absent from all cells, 1. less than 2 longitudinal septa (submuriform), 2. 3-6 longitudinal septa (muriform), 3. more than 6 longitudinal septa (muriform). (32) Orcinol depsides: 0. absent, 1. present. (33) Orcinol depsidones: 0. absent, 1. present. (34) Orcinol depsides or orcinol depsidones : 0. absent, 1. present. (35) β-Orcinol depsides: 0. absent, 1. present. (36) β-Orcinol depsidones: 0. absent, 1. present. (37) β-Orcinol depsides or β-orcinol depsidones: 0. absent, 1. present. (38) Xanthones: 0. absent, 1. present. (39) 6-O-methylarthothelin: 0. absent, 1. present. (40) Arthothelin: 0. absent, 1. present. (41) Usnic acid and related substances: 0. absent, 1. present. (42) Placodiolic acid: 0. absent, 1. present. (43) Anthraquinones: 0. absent, 1. present. (44) Aeruginose, HNO3+ violet exciple pigment (cinereorufa-green): 0. absent, 1. present. (45) Yellow, K+ red pigment in thallus or medulla: 0. absent, 1. present. 487 BIOGRAPHICAL SKETCH Frank Bungartz was born in Rheinbach, Germany on the 26th of October 1967. He received his elementary education at the Franziskusschule in Euskirchen, Germany and completed his secondary education at the Gymnasium Marienschule in Euskirchen. In 1988, Frank enrolled at Bonn University, majoring in Biology and Geography. He received his bachelor (Vordiplom) in 1991. During his graduate studies at Bonn University, Frank received an ERASMUS grant from the European Union to spent one year as an exchange student at the University of East Anglia, Norwich, U.K. Upon return, Frank began his research project on the lichen and vascular plant vegetation of the Brodenbach Valley, Mosel, Germany. He finished his master’s thesis (Diplomarbeit) in 1996 receiving his Master’s of Science (Diplom) from Bonn University. The following three years, Frank acquired professional experience as an ecological consultant, specializing in lichen ecology. In 1999 he was invited by Dr. Thomas H. Nash III to begin a PhD at Arizona State University. During his studies at Arizona State University, Frank became an active participant of the Sonoran Desert Lichen Flora Project, working both as an editor and contributor to the Lichen Flora of the Greater Sonoran Desert Region.

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The lichen genus Buellia de Not. in the greater Sonoran Desert Region: saxicolous species with one-septate ascospores

The lichen genus Buellia de Not. in the greater Sonoran Desert Region: saxicolous species with one-septate ascospores

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