Fungal Diversity (2011) 51:103–134 DOI 10.1007/s13225-011-0145-6 Capnodiaceae Putarak Chomnunti & Conrad L. Schoch & Begoña Aguirre-Hudson & Thida W. Ko-Ko & Sinang Hongsanan & E. B. Gareth Jones & Rampai Kodsueb & Rungtiwa Phookamsak & Ekachai Chukeatirote & Ali H. Bahkali & Kevin D. Hyde Received: 17 October 2011 / Accepted: 18 October 2011 / Published online: 16 November 2011 # Kevin D. Hyde 2011 Abstract In this paper we revisit the Capnodiaceae with notes on selected genera. Type specimens of the ascomycetous genera Aithaloderma, Anopeltis, Callebaea, Capnodaria, Echinothecium, Phragmocapnias and Scorias were re-examined, described and illustrated. Leptoxyphium is anamorphic Capnodiaceae and Polychaeton is a legitimate and earlier name for Capnodium, but in order to maintain nomenclatural stability we propose that the teleomorphic name should be considered for the approved lists of names currently in preparation for fungi. Notes are provided on the ascomycetous genus Scoriadopsis. However, we were unable to locate the type of this genus during the time frame of this study. The ascomycetous genera Aithaloderma, Ceramoclasteropsis, Hyaloscolecostroma and Trichomerium are excluded from Capnodiaceae on the basis of having ascostromata and trans-septate hyaline ascospores and should be accommodated in Chaetothyriaceae. Callebaea is excluded as the ascomata are thyriothecia and the genus is placed in Micropeltidaceae. Echinothecium is excluded as synonym of Sphaerellothecium and is transferred to Mycosphaerellaceae. The type specimen of Capnophaeum is lost and this should be considered as a doubtful genus. The coelomycetous Microxiphium is polyphyletic, while the status of Fumiglobus, Polychaetella and Tripospermum is unclear. Fourteen new collections of sooty moulds made in Thailand were isolated and Electronic supplementary material The online version of this article (doi:10.1007/s13225-011-0145-6) contains supplementary material, which is available to authorized users. P. Chomnunti : T. W. Ko-Ko : S. Hongsanan : R. Phookamsak : E. Chukeatirote : K. D. Hyde (*) Institute of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai 57100, Thailand e-mail: kdhyde3@gmail.com P. Chomnunti : T. W. Ko-Ko : S. Hongsanan : R. Phookamsak : E. Chukeatirote : K. D. Hyde School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand C. L. Schoch National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, 45 Center Drive, MSC 6510, Bethesda, MD 20892-6510, USA B. Aguirre-Hudson Jodrell Laboratory, Royal Botanic Gardens, Kew, Richmond, Surrey TW93DS, UK E. B. G. Jones Institute of Ocean and Earth Sciences (IOES), C308, Institute of Postgraduate Studies Building, University of Malaya, 50603, Kuala Lumpur, Malaysia R. Kodsueb Faculty of Science and Technology, Pibulsongkram Rajabhat University, Phisanulok 65000, Thailand A. H. Bahkali : K. D. Hyde Botany and Microbiology Department, King Saud University, College of Science, P.O. Box: 2455, Riyadh 1145, Saudi Arabia 104 sequenced. The nuclear large and small rDNA was partially sequenced and compared in a phylogeny used to build a more complete understanding of the relationships of genera in Capnodiaceae. Four new species are described and illustrated, while Phragmocapnias and Scorias are epitypified with fresh collections. Keywords Chaetothyriaceae . Micropeltidaceae . Molecular phylogeny . Sooty moulds Introduction The family Capnodiaceae was introduced by von Höhnel (1910) with the generic type Capnodium Montagne (1849) and presently includes 14 genera and 117 species (Kirk et al. 2008; Lumbsch and Huhndorf 2010). Capnodiaceae are sooty moulds with bitunicate asci borne in ostiolate ascomata; the family however is based mostly on ecological characters (von Arx and Müller 1975). The first complete monographic review of capnodiaceous sooty moulds recognized both sexual and asexual species in Eucapnodiaceae (Fraser 1935). Batista and Ciferri (1963a, b) later provided a monograph of Capnodiaceae in the order Capnodiales. Hughes (1972) reviewed and re-classified Capnodiaceae which was characterized by the structure of the hyphae, the presence or absence of pseudoparaphyses and by deviating conidial states. Members of this family also had superficial ascomata with ovoid asci in fascicles and hyaline to dark, one to multiseptate ascospores (see also Hughes 1976). More recently Capnodiaceae have been redefined by the following features: superficial mycelium of interwoven, mucilaginous, brown, cylindrical or tapering hyphae, mostly constricted at the septa, and occur as leaf epiphytes associated with the honeydew of insects (Hughes 1976; Andrew 1982; Blakeman and Fokkema 1982), the ascomata lack pseudoparaphyses and the asci are bitunicate. The fungi are also known as sooty moulds, and tend to live in complex communities, often with multiple fungal parasites, inhabiting a common sooty mass (Faull et al. 2002; Hughes 2003). They are noted for the production of darkly pigmented hyphae, often of very characteristic morphology (Hughes 1976; Reynolds 1998). Anamorphs reported in Capnodiaceae are Acanthorus, Apiosporium, Conidiocarpus, Conidioxyphium, Fumagospora, Fumiglobus, Leptoxyphium, Mycogelidium, Phaeoxyphiella, Polychaetella, Polychaeton, Scolecoxyphium, and Tripospermum (Hyde et al. 2011). The Capnodiaceae should not be confused with Chaetothyriaceae which are also referred to as sooty moulds since they share the same ecological niche and are similar in appearance. The main differences are found in the characteristics of the ascomata, being single locules in Fungal Diversity (2011) 51:103–134 Capnodiaceae, and ascostromata, often with more than one locule in Chaetothyriaceae. In addition, phylogenetic analyses have clearly shown them to be unrelated and they were placed in two separate classes: Dothideomycetes and Eurotiomycetes respectively (Schoch et al. 2007, 2009; Geiser et al. 2006; Chomnunti et al. 2012). The easiest character by which to distinguish these families on leaf surfaces is the form of the ascomata. In Capnodiaceae ascomata are subglobose to globose, with or without setae (von Arx and Müller 1975), while in Chaetothyriaceae they are ascostromata surrounded by a pellicle of superficial mycelium (Chomnunti et al. 2012). The purpose of this study is to revisit the family Capnodiaceae by examining available generic types which are described and illustrated. We have also collected fresh specimens from Thailand and we have isolated the material in culture and sequenced the species. We incorporate 14 new taxa in the molecular analysis and provide a more complete tree than has been presented before for Capnodiales (Crous et al. 2009). Material and methods Generic type specimens of Aithaloderma, Anopeltis, Callebea, Capnodium Echinothecium, Phragmocapnias and Scorias were obtained from Herbaria S, B, NY and K, and the collections are documented morphologically. The type material of some species could not be located during the time frame of this study; however, they are discussed based on the original description and subsequent publications. The herbarium specimens were rehydrated in 5% KOH for 10 min and free hand sections prepared under the stereo microscope, mounted in water and later preserved in lactic acid. Microcharacters were examined and observed under a compound microscope (Nikon 80i) fitted with DIC, and measurements made with the Tarosoft (R) Image Frame Work. Isolates and morphology Sites were visited throughout the provinces of Chiang Mai and Chiang Rai in northern Thailand and leaves of various plants with sooty mould-like colonization were collected and brought to the laboratory in plastic bags. Sections of ascomata were made free hand and mounted in lactoglycerol. Melzer’s reagent was used to test the amyloidity of the apical ring and Indian ink was used for demonstrating the mucilaginous sheath (before adding the lactoglycerol). Pure cultures were obtained by single spore isolation. For that, a part of the hymenium containing ascospores was removed from 4 to 5 ascomata of fresh material, using a sterile needle, and placed in a drop of Fungal Diversity (2011) 51:103–134 sterile water on a glass slide. The contents were broken up further mechanically with the sterile needle until a spore suspension was obtained. The spore suspension was then transferred with a sterile pipette onto the surface of a Petri dish with 2% Difco potato-dextrose agar (PDA) as media. The plates were left overnight to germinate and observed within 12 h. Germinating spores were individually transferred onto a fresh Petri dish. Isolates were grown on PDA at 28 C for 12 h of light/12 h of dark for routine maintenance. Colony colour and characteristics were assessed after 4 weeks, and this material was used for molecular study. Types of isolated new species are deposited at the Mae Fah Luang University (MFLU) Herbarium, Chiang Rai, Thailand, and the cultures in the Culture Collection of the same institution (MFLUCC) and in BIOTEC Culture Collection (BCC). DNA isolation, amplification and sequencing The analysis was performed with two markers: partial sequences from the small and large subunits of the nuclear ribosomal RNA genes (SSU, LSU). The individual genes were aligned with SATé (Liu et al. 2009) using MAFFT (Katoh et al. 2009) as the external sequence alignment tool and RAxML (Stamatakis 2006) as the tree estimator. Representative sequences from Capnodiales were downloaded from GenBank according to Supplementary Table 1. Dothidea insculpta and D. sambuci were selected as outgroups. Phylogenetic analyses of the single genes did not yield conflicts in clades with RAxML bootstrap presence above 70% (data not shown), therefore sequences were concatenated in BioEdit (Hall 2004). This resulted in 15.5% missing and gap characters out of a total set of 1982 characters (967 obtained from SSU and 1,015 obtained from LSU). The final data matrix had 51 taxa including outgroups. A phylogenetic analysis was performed at the CIPRES webportal (Miller et al. 2010) using RAxML v. 7.2.8 as part of the “RAxML-HPC2 on TG” tool (Stamatakis 2006; Stamatakis et al. 2008). A general time reversible model (GTR) was applied with a discrete gamma distribution and four rate classes. Fifty thorough maximum likelihood (ML) tree searches were done in RAxML v. 7.2.7 under the same model, each one starting from a separate randomised tree and the best scoring tree selected with a final ln value of −7912.128405. One thousand non parametric bootstrap iterations were run with the GTR model and a discrete gamma distribution. The resulting replicates were plotted on to the best scoring tree obtained previously. The phylogram with bootstrap values above the branches is presented in Fig. 1 by using graphical options available in TreeDyn v. 198.3 (Chevenet et al. 2006). 105 We also analyzed the same data set mentioned above using the Bayesian method of Huelsenbeck et al. (2001) by implementing Markov Chain Monte Carlo (MCMC) sampling using the software MrBayes v.3.1.2 (Huelsenbeck and Ronquist 2001) at the CIPRES webportal. Data were analysed with a GTR model with gamma-distributed rate variation across sites (invariance, partitioning across genes) were applied with four discrete gamma categories. The Bayesian prior distributions treated all trees as equally likely and two parallel runs were performed with four chains each. This was continued for 2,000,000 generations and every 100th tree was saved. The two runs were verified for convergence and the first 50% of each run was discarded as burn in with the remaining two sets of 10,000 trees combined. The best scoring RAxML tree was used as a template and the percentage presence of all nodes within the combined set of 20,000 trees plotted on its nodes as Bayesian posterior probabilities. These are indicated as the second set of values (after the front slash) in Fig. 1 Results Molecular phylogeny In the tree (Fig. 1) we included representative sequences of Capnodiaceae, Davidiellaceae, Dissoconiaceae, Mycosphaerellaceae, Schizothyriaceae, Teratosphaeriaceae as well as uncertainly placed lineages in Capnodiales mainly following an earlier analysis by Crous et al. (2009). All groups agreed with this analysis. The phylogeny presented includes the biggest sampling of Capnodiaceae to date and includes several samples from earlier studies (Crous et al. 2009; Reynolds 1998; Ruibal et al. 2009; Schoch et al. 2007). The 15 specimens of sooty moulds from Thailand including Leptoxyphium cacuminum, Phargmocapnias betle, P. asiaticus, P. siamensis, Polychaeton coartatum and Scorias spongiosa clustered within Capnodiaceae (Fig. 1). The clade containing Capnodiaceae was only recovered in 50% of bootstrapped (BP) tree using the RAxML analysis but received a 99% posterior probability (PP) in the Bayesian analysis. This clade is recovered with high frequency in other analyses including protein coding gene markers (Schoch et al. 2009) but not well supported in other studies using only ribosomal sequence comparisons (Crous et al. 2009). The Capnodiaceae comprises four clades (A–D). Clade A (100% BP and PP) comprises three Leptoxyphium sequences from GenBank and three new strains with characters typical of Leptoxyphium sensu Hughes (1976), Olejnik et al. (1999) and Cheewangkoon et al. (2009) as well as Microxiphium citri and a rock isolate with 97% support. Microxiphium citri was isolated from citrus fruit 106 Fungal Diversity (2011) 51:103–134 Fig. 1 A RAxML maximum likelihood tree from obtained from a data set of 51 taxa including representatives of Capnodiales, focused on Capnodiaceae, comparing two genes (SSU, LSU). The first set of numbers above the nodes are bootstraps from 1000 pseudorepetitions and the second represent Bayesian posterior probabilities expressed as percentages. Only values above 50% are shown. Culture and voucher numbers are indicated after species names and names of isolates with newly obtained sequences for this study are bolded in Spain and may be wrongly named. Clade B (52% BP, 92% PP 100%)) comprises Capnodium coffeae, Conidioxyphium gardeniorum, Microxiphium aciculiforme and a new species of anamorphic Capnodium (= Polychaeton). Conidioxyphium gardeniorum is either wrongly named or is a synonym of Capnodium. Clade C are Phragmocapnias (= Conidiocarpus) species with one teleomorph species being identical to the generic type and is thus epitypified. The genus is represented by at least two morphological species which are described as new in this paper. Further collections and analyses are likely to reveal other taxa in this genus. Basal to the Phragmocapnias Clade C is Polychaeton citri, a strain of Antennariella placitae, Microxyphium theae and two ant nest wall strains. Polychaeton citri was isolated from a leaf of Citrus aurantium together with Pseudococcus citri (Crous et al. 2009). Because of difficulty in isolating Capnodium species and the fact that many species occur on the same leaf in close proximity we suspect this may be Fungal Diversity (2011) 51:103–134 wrongly named and this will have to be verified. Antennariella placitae was described by Cheewangkoon et al. (2009) but it is not clear if this is correctly placed and more collections would be needed to establish its familial relationships. Sister to these taxa is a putative strain of Capnodium salicinum which also require verification. Finally, Clade D (100% bootstrap support) comprises two isolates of Scorias spongiosa. The new isolate from Thailand is designated as the epitype here. Three Microxiphium species are included in the analysis, which Fig. 2 a–e Sooty moulds growing on host leaves. a Unidentified tree. b Bischofia javanica. c Euphorbia sp. d Psidium guajava. e Cestrum diurnum. f–i Representative pycnidia of Capnidiaceae genera accepted in this paper. f Phragmocapnias. g Capnodium. h Leptoxyphium. i Scorias 107 in the present study show that the genus is polyphyletic. Based on morphological examination of the type species, this genus may belong in Coccodiniaceae. Capnodiaceae (Sacc.) H hn. ex Theiss. 1916. Saprobic on honey dew excretions from insects, usually forming blackened sooty-like regions on green healthy leaves, stems, bark and even rocks (Fig. 2a–e), and often in association with numerous other species and fungicolous taxa. Thallus comprising mycelium with black sooty 108 growth on the host surface, producing superficial colonies with septate, cylindrical, brown hyphae. Ascomata arise from the mycelial mass, subglobose to globose, coriaceous, with or without setae, dark brown, with a central ostiole. Peridium brown, relatively thin, comprising cells of textura angularis. Pseudoparaphyses lacking. Asci bitunicate, saccate, with a short pedicel, lacking an ocular chamber. Ascospores multiseptate or muriform, hyaline to brown. Anamorph states are pycnidial, mostly elongated, with short or long narrow necks, and with or without a swelling where the conidia are produced. Ostiole prominent at the apex of the pycnidial neck. Conidia occur on typical capnodiaceous hyphae within the pycnidia and are small, one-celled, ellipsoidal, and hyaline. Family type: Capnodium Mont. (1849) (= Polychaeton (Pers.) Lév. 1846). The Melbourne Botanical Congress has recently approved large scale changes to process of naming fungi, including the abolition of Article 59. Therefore, by applying the principle of priority Polychaeton would be the earlier and valid name for the species currently assigned to the genus Capnodium. However, this process is still being finalized and it was proposed that anamorph typified names should not been taken up to displace teleomorph typified names in use until each case has been reviewed by the Fungal Committee established at the Melbourne Congress (Hawksworth 2011). As the name Capnodium is taken up for the name of the ranks of family and order in this group of fungi, this is a prime example that the retention of the name should be proposed for approval to the Committee. The family Capnodiaceae was introduced by von Höhnel (1909) and validated by Theissen in 1916 according to von Arx and Müller (1975) and Hughes (1976), and is circumscribed as dothideomycetous sooty moulds with mostly pycnidial anamorphs (Reynolds 1982). The taxa of this family can be recognized by black mycelial growth spreading on the host surface which produces superficial colonies with septate, cylindrical, dark-brown hyphae (Fig. 3a, b). Ascomata arise from the mycelial mass and are subglobose to globose (Sivanesan 1984; Barr 1987; Figs. 2e and 3c). Their anamorphic states, placed mostly in Asbolisiaceae by Batista and Cifferi (Batista and Ciferri 1963a, b) form elongated pycnidia, with short or long narrow necks, an apical ostiolar canal, and minute, one-celled and hyaline conidia (sensu Hughes 1976). In the past some members of the family had been only known from anamorphic stages, and these pycnidia had been assigned to one or other teleomorphic genera and families depending on whether the researchers gave more emphasis to Saccardoan generic concepts such as spore morphology and colour, or colony hyphal development, hyphal morphology and ascomatal and pycnidial formation (as with Hughes 1976), but now the use of modern Fungal Diversity (2011) 51:103–134 molecular techniques has enabled us to confirm their relationship in the Capnodiaceae. Crous et al. (2009) used molecular methods to evaluate the monophyly of the order Capnodiales, yet they did not discuss the family in detail, though concluding that it probably contained diverse lineages. In the present account we illustrate genera of the group and we compare our morphological findings of some generic types examined with molecular results of taxa found in south eastern Asia. Key to genera accepted in Capnodiaceae 1. Pycnidia with a black stalk, and an upper brown swollen region producing conidia; ascospores hyaline Phragmocapnias (Conidiocarpus) (Fig. 2f) 1. Pycnidia without a black stalk or an upper brown swollen region producing conidia; ascospores brown or hyaline2 2. Pycnidia lacking a basal bulbous part; ascospores brown Capnodium (Polychaeton) (Fig. 2g) 2. Pycnidia with basal bulbous part; ascospores other 3 3. Ascomycetous state unknown, pycnidia with narrow bulbous base, apex cup-like which produces conidia Leptoxyphium (Fig. 2h) 3. Ascomata present, gregarious on blacked mycelial mat; pycnidia with swollen bulbous base 4 4. Ascospores mostly 3-septate. Scorias (Fig. 2i) 4. Ascospores mostly 1-septate. Scoriadopsis Phragmocapnias Theiss. & Syd., Ann mycol. 15(6): 480 (1918) [1917] Conidioxyphium Bat. & Cif., Quad. Lab crittogam., Pavia 31: 72 (1963) Podoxyphium Speg., Physis. B. Aires 4: 294 (1918) Conidiocarpus Woron., Ann mycol. 24 (3/4): 250 (1926) Saprobic on sugary exudates from insects, dark mycelium forming a soot-like coating on the upper surface of leaves. Thallus composed of black, pelliculose, reticulately branched, dense, cylindrical, radiating, septate hyphae. Ascomata scattered, subglobose to broadly ellipsoidal, barely stalked, firmly attached to the basal hyphae, dark brown, thick-walled, ostiolate, with setae. Peridium consisting of pale to dark brown cells forming a textura angularis. Asci bitunicate, 8-spored, broadly clavate, with short pedicle. Ascospores cylindric-clavate, hyaline, 4-septate and constricted at the septum. Pycnidia with a black stalk, with an upper brown swollen part which produces conidia, wall comprising cylindrical cells. Ostiole surrounded by hyaline hyphae. Conidiogenous cells form from inner cell surface of swollen part. Conidia small, ellipsoid, 1-celled, hyaline, smooth-walled. Anamorph: Conidiocarpus Woron. (Hyde et al. 2011) Fungal Diversity (2011) 51:103–134 Type species Phragmocapnias betle (Syd. P. Syd. & E.J. Butler) Theiss. & Syd., Ann mycol. 15(6): 480 (1918) [1917] ≡ Capnodium betle Syd. P. Syd. & E.J. Butler., Ann Mycol 9 (4): 384 (1911) Description from holotype (Fig. 3a–i) Saprobic on sugary exudates from insects growing on the surface of living leaves (Fig. 3a). Thallus thin, amphigenous, black, pelliculose, composed of reticulately branched, dense, cylindrical to somewhat constricted, radiating, septate hyphae (Fig. 3b). Ascomata up to 109 101–110 μm diam, 99–111 μm high (x ¼ 106  105 mm, n= 5), scattered, subglobose to broadly ellipsoidal, with short stalks, firmly attached onto the radiating basal hyphae, dark brown, thick-walled, ostiole present in mature ascomata, setae with blunt apices scattered over the ascomata (Fig. 3c, 3). Peridium 15–19 μm wide (x ¼ 17 mm, n=10), composed of pale to dark brown, cells arranged in a textura angularis (Fig. 3e). Asci 37–48×17– 30 μm (x ¼ 42  22 mm, n = 5), 8-spored, bitunicate, broadly clavate, with short pedicle (Fig. 3f–h). Ascospores 18–27×5 μm (x ¼ 21  5 mm, n=5), fasciculate, cylindricclavate, ends rounded, upper cells slightly wider than the lower cells, hyaline, 4–septate, constricted at the septa, smooth-walled (Fig. 3f–i). Fig. 3 a–i Phragmocapnias betle (holotype of Capnodium betle). a, b Ascomata on leaves. c, e Vertical section through stalked ascoma. d Ascomata exterior wall with setae. f–h Asci with short pedicel. i Cylindric–clavate ascospores. Bars: e=100 μm, c, d=50 μm, f, h, g=20 μm, i=10 μm 110 Description from epitype (Fig. 4a–i) Saprobic on sugary exudates from insects growing on the upper surface of living leaves (Fig. 4a). Thallus thin, amphigenous, black, pelliculose, composed of reticulately branched, dense, cylindrical to somewhat constricted, radiating, septate hyphae (Fig. 4a). Ascomata 82–93×84–105 μm diam (x ¼ 88  92 mm, n=10), superficial, subglobose to broadly ellipsoidal, dark brown to black, shiny, coriaceous, Fungal Diversity (2011) 51:103–134 with 3–5 setae surround the ostiole setae 50–75×3–5 μm (x ¼ 57  4 mm, n=10), dark brown, septate, tapering with rounded ends (Fig. 4a, b). Ostiole present in mature ascomata. Peridium thin, comprising cells arranged in a textura angularis. Asci 43–53×13–33 μm (x ¼ 24  21 mm, n=10), 8-spored, bitunicate, fusiform to broadly clavate, with a short pedicel, short ocular chamber observed in young asci (Fig. 4d, e). Ascospores 20–24 × 4.8–5.8 μm (x ¼ 23  5:3 mm, n=10), fasciculate, cylindric-clavate, ends rounded, upper cells slightly wider than the lower cells, Fig. 4 a–l Phragmocapnias betle (epitype). a Ascomata on living leaf. b, c Ascomata which are subglobose with setae. d, f Bitunicate asci. e Mycelium beneath ascomata. g–l Ascospores surrounded by a mucilaginous sheath. Bars: b, c=100 μm, d, f=50 μm, e=20 μm, g–l=10 μm Fungal Diversity (2011) 51:103–134 hyaline, 4 septate, constricted at all septa, guttulate, some surrounded by a mucilaginous sheath (Fig. 4f–l). Material examined: BANGLADESH, Dhaka, on leaves of Piper betle, 5 April 1910, A.L. Som 1061 (S, holotype of Capnodium betle); THAILAND, Chiang Rai Province, Mae Fah Luang University, on living leaf of Ixora sp., 24 April 2009, Putarak Chomnunti, DPC 014 (MFLU09-0650, epitype designated here), extype living culture in MFLUCC10-0053; Ibid., on living leaf of Mimusops elengi Linn., 14 July 2009, Putarak Chomnunti (MFLU 09-0647), living culture in MFLUCC10-0050. Phragmocapnias betle has been reviewed by Reynolds (1979) who recognizes this genus as having stalked ascomata with setae and hyaline ascospores with trans-septa. In MFLU 09-0647 the ascomata are slightly bigger than in the type: 87–137× 80–125 μm diam. [x ¼ 103:5  94:5 mm, n= 10], but the size of asci and ascospores fit the range given in the above descriptions: 42–50×10–15 μm [x ¼ 41  13 mm, n= 10] and 20–25× 5–8 μm [x ¼ 22  5 mm, n= 20] respectively. Phragmocapnias has been discussed fairly extensively and there are eight names listed in Index Fungorum of which five are given as congeneric (see http://indexfungorum.org/Names/ Names.asp). Reynolds (1978) however, concluded that there was only one species, P. betle and excluded the other species based on various reasons. The anamorph of P. betle was reported to be Conidiocarpus (Hughes 1976), but Reynolds (1979) concluded that the anamorph of Scorias and Phragmocapnias were uncertain. However, the phylogenetic data presented here (Fig. 1) confirm that Conidiocarpus is the anamorph of Phragmocapnias. In the combined gene phylogenetic analysis the epitype and second collection of Phragmocapnias betle clustered in Capnodiaceae and are basal to two species (6 strains) of anamorphic Phragmocapnias (= Conidiocarpus) confirming the anamorph-teleomorph linkage. The eight strains of Phragmocapnias used in this study cluster together with 75% bootstrap support with the two Phragmocapnias species closest with 99% bootstrap support. Phragmocapnias asiaticus is closest to the Phragmocapnias species and the five strains of Phragmocapnias siamensis cluster above with 81% bootstrap support indicating that they may be more than one species. Conidiocarpus penzigii Woron. (as penzigi), Ann mycol. 24(3/4): 250 (1926). Saprobic on sugary exudates from insects forming a sootylike coating on the upper surface of living leaves, which crumbles away easily when dry. Thallus of blackish-gray, comprising superficial mycelia 5–6 μm wide, with cylindrical, pale brown cells, constricted at the septum, anastomosing. The generic type of Conidiocarpus is C. penzigii Woron. which is saprobic on sugary exudates from insects forming 111 a sooty-like coating on the upper surface of living leaves, which crumbles away easily when dry (Woronichin 1926). Pycnidia are reported to be 420–500 μm high, arising from mycelial mass, olive-brown, stalk 270-335 μm long, 2330 μm thick, blackish, swollen part 100×33 μm, comprising short, olive-brown, angular cells, neck 50–66×13 μm, ostiole surround by hyaline hyphae up to 20 μm long. Conidia are 5–9×1.5 μm, oblong-ellipsoid, hyaline, unicellular (Woronichin 1926). We have been unable to locate the type of C. penzigii which is described from Georgia. Conidiocarpus however, is a common sooty mould and we made several collections in Thailand. Presently only two species are listed in the genus, C. penzigii (Woronichin 1926) and C. longicollus Matsush (Matsushima 2003). Conidioxyphium (2 species) and Podoxyphium (16 records) are considered as synonyms and the group needs further study. Below we introduce two new species of Phragmocapnias as anamorphic states with molecular data which differ from existing Conidiocarpus species in conidia size. We also transfer C. penzigii (Woronichin 1926) and C. longicollus Matsush to Phragmocapnias. Phragmocapnias asiaticus Chomnunti & KD Hyde, sp. nov. (Fig. 5a–k) MycoBank: 563360 Conidiocarpus penzigii Woron. similis sed conidia 2.5– 3.7×1–1.4 μm differt. Etymology: ‘asiaticus’ in reference to it origin in Asia. Habit forming a soot-like coating on the upper surface of leaves (Fig. 5a). Thallus superficial, consisting of a network of cylindrical and septate, 2.2–3.7 μm, thick (x ¼ 3 mm, n=30) hyphae, constricted at the septum, pale brown, but dark brown towards the edge (Fig. 5c, d). Pycnidia 302–387(−471) μm high (x ¼ 366 mm, n=10), arising from capnodiaceous type hyphae, black at the base and stalk, 17– 25 μm wide (x ¼ 21 mm, n=10), the conspicuous oval swelling which produce conidia is 18–24 μm wide (x ¼ 21 mm, n=10) and brown, comprising of cylindrical septate cells (Fig. 5e–g). Ostiole 4.9–6.5 (x ¼ 5:6 mm, n=10), surrounded by hyaline hyphae (Fig. 5h). Conidiogenous cells formed in the inner cells of the oval part. Conidia 2.5–3.7× 1–1.4 μm (x ¼ 3:1  1:2 mm, n=30), oblong to ellipsoid, 1-celled, hyaline, rounded ends (Fig. 5i–k). Material examined: THAILAND, Chiang Rai Province, Doi Tung, on living leaf of Coffea arabica, 15 August 2009, Jian Kui Liu, DPC 027 (MFLU09-0660, holotype), extype living culture in MFLUCC10-0062. Phragmocapnias longicollus (Matsush.) Chomnunti & KD Hyde, comb. nov. ≡ Conidiocarpus longicollus Matsush. Matsush. Mycol. Mem. 10: 85 (2003) [2001] MycoBank: 374479 112 Fungal Diversity (2011) 51:103–134 Fig. 5 a–k Phragmocapnias asiaticus (holotype). a Black mycelium covering the leaf surface. b Pycnidium with long stalks on the host. c, d Mycelial network. e, f Conical pycnidium and pycnidium wall. Note the conida in the swollen part in f. g Pycnidia. h Ostiole. i–k Conidia. Bars: c, e=50 μm, d–f=20 μm, h, k=10 μm, i, j=5 μm Phragmocapnias penzigii (Woron.) Chomnunti & KD Hyde, comb. nov. ≡ Conidiocarpus penzigii Woron. [as penzigi], Ann mycol. 24(3/4): 250 (1926). MycoBank: 273921 Phragmocapnias siamensis Chomnunti & KD Hyde, sp. nov. (Fig. 6a–g) MycoBank: 563361 Conidiocarpus penzigii Woron. similis sed conidia 4.5– 5.2×1.9–2.4 μm differt. Etymology: from siamensis in reference to the Latin name of Thailand, where the species originated. Saprobic on sugary exudates from insects growing on the upper surface of living leaves forming soot-like coating. Thallus of 3–5 μm thick (x ¼ 4 mm, n=20) hyphae, which is superficial, cylindrical, septate, constricted at the septum and pale brown (Fig. 6c, d). Pycnidia brown, comprised of Fungal Diversity (2011) 51:103–134 113 Fig. 6 a–g Phragmocapnias siamensis (holotype). a Black mycelium covering the leaf surface. b Pycnidia on host. c, d Mycelial network. e, f Conical pycnidia and pycnidia wall. g Conidia. Bars: f=100 μm, c–e=50 μm, g=20 μm cylindrical septate cells, 378–458 μm high (x ¼ 408 mm, n= 10), stalk black, 22–31 μm high (x ¼ 27 mm, n=10) at the base, the oval part produces conidia, 36–41 μm wide (x ¼ 38 mm, n=10) (Fig. 6e, f). Ostiole 9–15 (x ¼ 12 mm, n=10), surrounded by hyaline hyphae. Conidiogenous cells formed on the inner cell walls of the oval part. Conidia 4.5– 5.2×1.9–2.4 μm (x ¼ 4:8  2:1 mm, n=20), ellipsoid, aseptate, hyaline, smooth walled, ends rounded (Fig. 6g). Material examined: THAILAND, Chiang Rai Province, Thasud, on living leaf of Mangifera indica, 20 August 2009, Rungtiwa Phokhomsak, DPC 029 (MFLU09-0662, holotype), extype living culture in MFLUCC10-0064; Ibid., Bandoo, on living leaf of Mangifera sp., 7 June 2009, Putarak Chomnunti, DPC 026 (MFLU09-0656), living culture in MFLUCC10-006; Ibid., Doi Tung, on living leaf of Coffea arabica, 15 August 2009, Jian Kui Liu, DPC 028 (MFLU09-0661), living culture in MFLUCC10-0063; Ibid., on living leaf of Euphorbia sp., 15 August 2009, SC Karunarathna, DPC 030 (MFLU09-0663), living culture in MFLUCC10-0065; Ibid., Mae Fah Luang University, on living leaf of Bischofia javanica, 9 January 2010, Putarak Chomnunti, DPC 036 (MFLU10-0003), living culture in MFLUCC10-0074. Leptoxyphium Speg., Physis, B. Aires 4: 294 (1918) Astragoxyphium Bat., Nascim. & Cif., in Batista & Ciferri, Quad. Lab, crittogam., Pavia 31:45 (1963) Caldariomyces Woron., Ann mycol 24 (3/4): 261 (1926) Megaloxyphium Cif., Bat. & Nascim., Publcoes Inst. Micol. Recife 47:3 (1956) Saprobic on sugary exudates from insects growing on the surface of living leaves. Thallus of superficial, grey brown to brown, septate, branched mycelium, constricted at the septa, forming an irregular network. Pycnidia superfi- 114 cial, gregarious, arising from aggregated hyphae, base bulbous, comprising parallel hyphae, straight to slightly flexuous, sometimes with helical twisting, comprised of cylindrical hyphae and expanded to become funnel-shaped, capulate at the apex. Conidia ellipsoidal, hyaline, 1-celled, guttulate (Woronichin 1926; Hughes 1976). Type species Fungal Diversity (2011) 51:103–134 Mycelium form an irregular network which comprises cylindrical hyphae. Pycnidia arising from helically twisting hyphae or ropes of repent hyphae, stalk with a stout base, mostly unbranched, forming a fringe of sterile hairs at the apex. Conidiogenous cells formed on the inner cell walls of the swollen apex. Conidia usually broadly ellipsoidal, some 1-septate, and pigmented when forming a mass on the host surface (Saccardo and Sydow 1899; Hughes 1976). Leptoxyphium graminum (Pat.) Speg., Physis, B. Aires 4:294 (1918) ≡ Capnodium graminum Pat., J. Bot., Paris 11: 348 (1897) Leptoxyphium cacuminum Chomnunti & KD Hyde, sp. nov. (Figs. 7a–l and 8a–l) MycoBank 563359 Fig. 7 a–l Leptoxyphium cacuminum (holotype). a Gregarious pycnidia on host surface. b, d, g Stalked pycnidia with wider base. e Formation of pycnidia from aggregated hyphae. c, f Black stalked funnel cupulate apex. h–l Conidia, conidiogenous boundary with hyaline hyphae surrounding the ostiole. Bars: b, d, e, g=200 μm, c–f= 50 μm, h–l=20 μm Fungal Diversity (2011) 51:103–134 115 Fig. 8 a–l Leptoxyphium cacuminum (holotype) in culture. a Colony on PDA. c, d Conidial mass at the apex of conidia. d, e Olive-green stalked funnel shaped at apex. f–h Septate hyphae. i–l Conidia produced from the apex of conidiophores. Bars: a=2 cm, d=200 μm, e–l=50 μm Leptoxyphium graminum (Pat.) Speg. similis sed conidia 4.1–6.7×2.1–2.7 μm differt. Etymology: from the Latin—cacumin meaning swollen, in relation to the pycnidia. 116 Saprobic on sugary exudates from insects growing on the surface of living leaves (Fig. 7a). Thallus of 3.2–6.2 μm high (x ¼ 4:4 mm, n=20), grey brown to brown, septate, branched, superficial mycelium, constricted at the septa, forming an irregular network. Pycnidia 341–446 μm high (x ¼ 392 mm, n=10), and 19–30 μm base (x ¼ 26 mm, n=10), superficial, gregarious, arising from aggregated hyphae, base bulbous, comprising parallel hyphae, straight to slightly flexuous, sometime with helical twisting (Fig. 7b–e). Stalked pycnidia comprising cylindrical hyphae expanding at the end into a funnel-shape, resembling a cupula, 27–43×28–45 μm [x ¼ 33  35 mm, n=10] (Fig. 7f–h). Conidiogenous cells arising from the inner cell wall of the cupulate apex. Conidia ellipsoidal, hyaline, aseptate and guttulate, 4.1–6.7×2.1– 2.7 μm [x ¼ 5:2  2:4 mm, n=20] (Fig. 7i–l). In culture colonies reaching to 5 cm diam. after 10 days growth on PDA medium. Colony flat, irregular in the middle but radiating towards the edge, dull black, becoming olive-green towards the edge (Fig. 8a–c). Mycelium of cylindrical and septate hyphae 3.6–4.8 μm [x ¼ 4:2 mm, n=20], becoming aerial, branched, pale brown to deeply pigmented at the septum (Fig. 8f–h). Pycnidia stalked, arising from a basal cell and developing a cupulate swelling towards the apex. Stalk olive-green, and deeply pigment at the base, but at the apex the hyphae are hyaline (Fig. 8d, e). Conidia broadly ellipsoidal, unicellular and guttulated, hyaline, 4–4.9×3.5– 3.7 μm [x ¼ 4:4  3:6 mm, n=20] (Fig. 8i–l). Material examined: THAILAND, Chiang Rai Province, Thasud, on living leaf of Gossypium herbaceum, 11 August 2009, SC Karunarathna, DPC 024 (MFLU09-0657, holotype), extype living culture in MFLUCC10-0059; Ibid., Bandoo, on living leaf of Ficus sp., 4 January 2010, KD Hyde, DPC 050 (MFLU 10-0015), living culture in MFLUCC10-0086; Ibid., Mae Fah Luang University, on living leaf of Mimusops elengi, 24 April 2009, Putarak Chomnunti, DPC 009 (MFLU09-0646), living culture in MFLUCC10-0049. Sequences of Leptoxyphium fumago, L. madagascariense and L. kurandea are available in GenBank and Leptoxyphium cacuminum clearly clusters with them. Nevertheless, Leptoxyphium cacuminum differs from the other known species in the genus because of its hyaline conidia, never becoming septated or pigmented when mature. Capnodium Mont., Annls Sci. Nat., Bot., sér. 3 11: 233 (1849) Polychaeton (Pers.) Lév., in Orbigny, Dict. Univ. Hist. Nat. 8: 493 (1846) Capnodaria (Sacc.) Theiss. & Syd., Ann mycol. 15(6): 474 (1918) [1917] Capnodium sugen. Capnodaria Sacc., Syll. Fung. (Abellini) 1:74 (1882) Capnodenia (Sacc.) Theiss. & Syd. (1917) Fumago sect. Polychaeton Pers., Mycol. eur. (Erlanga) 1: 9 (1822) Fungal Diversity (2011) 51:103–134 Fumagospora G. Arnaud, Annals d’École National d’Agric. de Montpellier, Série 2 10(4): 326 (1911) Morfea Roze, Bull. Soc. bot. Fr. 14: 21 (1867) Morfea (G. Arnaud) Cif. & Bat., in Batista & Ciferri, Saccardoa 2: 153 (1963) Phaeoxyphiella Bat. & Cif., Quad. Lab. crittogam., Pavia 31: 145 (1963) Scolecoxyphium Cif. & Bat. Publções Inst. Micol. Recife 47: 5 (1956) Saprobic on sugary exudates from insects growing on the surface of leaves, fruits, stems and other non plant objects. Thallus a loose or dense network of pale brown, superficial hyphae or a thick pseudoparenchymatous stromata, with sexual and asexual states often growing together. Ascomata superficial on mycelium of thallus, brown to dark brown or black, globose to ellipsoidal, short stalked or sessile, ostiolate at maturity, scattered or in groups, without setae. Peridium comprising dark brown to pale brown, thick-walled cells forming a textura angularis. Asci 8-spored, bitunicate, clavate, ovoid or saccate, aparaphysate, apedicellate. Ascospores brown, oblong or ovoid and some reniform, transseptate with one or more vertical septa or without vertical septa. Pycnidia elongate, often with long stalks, dark brown. Ostiole at apex of pycnidia. hyphae continued upwards to the tapered neck, terminating in an ostiole which is surrounded by dull hyphal round ends. Conidia hyaline, 1-celled. Anamorph: Polychaeton (Pers.) Lév. (Hyde et al. 2011). Polychaeton was introduced by Léveillé (1847) based on Persoon’s Fumago [subgenus] Polychaeton, and Hughes (1976) provides a detailed account of the choice of generic type including the reason why Spegazzini’s lectotype P. carolinense (Berk. & Desm.) Speg. was inadmissible (not included by Léveillé at the time of description). It is not clear whether Léveillé intended to include F. citri in the genus or whether it was meant to represent another genus. Of the five species in Persoon’s subgenus Polychaeton, only P. citri or P. quercinum were considered suitable to be generic types by Hughes (1976), who chose P. quercinum (≡ Fumago quercina) as the lectotype species. However, and unlike Berkeley and Desmazières (1849), he regarded this genus as the anamorphic stage of the genus Scorias, not Capnodium. This has been discussed by other authors, e.g. in Sutton (1977) as Polychaeton, see Index Fungorum (http://www.indexfungorum.org/Names/NamesRecord.asp? RecordID=206987) as Capnodium, and as type unknown in MycoBank (see http://www.mycobank.org/MycoTaxo. aspx?Link=T&Rec=4305), or non designated as Index Nominum Genericoum (see http://www.botany.si.edu/ing/ genusSearchTextMX.cfm). In this study we accept P. quercinum as the generic type, but Persoon’s original specimen was not available on loan. However, we have seen a possible type or authentic collection in herbarium K, which is part of M.J. Berkeley’s herbarium. Fungal Diversity (2011) 51:103–134 Capnodium was introduced by Montagne (1849) based on Fumago citri Pers. and is the type genus of Capnodiaceae (Friend 1965). Reynolds (1978) however, examined type material from L which only had anamorphic characters and this was similar for the supporting specimens listed in exsiccatae. Reynolds (1978) therefore chose a species where the ascus and the ascospores are known, the second species listed in the genus by Montagne (1849): Capnodium salicinum Mont. as the lectotype (Reynolds 1999). Index fungorum (http://www.speciesfungorum.org/Names/ SynSpecies.asp?RecordID=164508), however, lists this as a synonym of C. citri which we follow here. Capnodium is the teleomorph stage of Polychaeton (Figs. 10, 11 and 12), but it is the earlier name. As mentioned earlier that we advocate that Capnodium be considered for conservation under the “one fungus one name” concept that will be incorporated in the newly proposed Code for Algae, Fungi and Plants. In this study we provide nine sequences of ‘polychaeton-like’ Capnodium specimens and illustrate three species. Type species Capnodium citri Berk. & Desm., in Berkeley, J Rl Hort Soc 4: 11 (1849) (Fig. 9a–g) ≡ Fumago citri Pers., Mycol. eur. (Erlanga) 1: 9 (1822). ≡ Polychaeton citri (Pers.) Lév., in Orbigny, Dict. Univ. Hist. Nat. 8: 493 (1846) ≡ Microxiphium citri (Berk. & Desm.) Speg., Boln Acad. nac. Cienc. Córdoba 26(2–4): 399 (1924) [1923] = Apiosporium citri Briosi & Pass., Atti R.acad. Lincei, Trans., Sér. 3 7: 22 (1882). = Apiosporium salicis Kunze, in Kunze & Schmidt, Mykologische Hefte (Leipzig) 1: 8 (1817) = Capnodium salicinum Mont., Annls Sci. Nat., Bot., sér. 3 11: 234 (1849) = Pleosphaeria salicina (Mont.) G. Arnaud = Polychaeton salicinum (Mont.) Kuntze, Revis. gen. pl. (Leipzig) 3: 1–576 (1891) = Teichospora salicina (Mont.) Gäum. = Limacinia citri (Briosi & Pass.) Sacc., Hedwigia 36: 20 (1897) = Meliola citri (Briosi & Pass.) Sacc., Syll. fung. (Abellini) 1: 69 (1882) Saprobic on sugary exudates from insects growing on the surface of leaves, fruits, stems and other non plant objects. Thallus comprising 4.2–6 μm wide, dark brown, superficial, cylindrical and septate hyphae, constricted at the septum (Fig. 9a, b, f). Pycnidia up to 345–391 long μm, 36–40 μm wide at the base, arising from dense mycelia, elongate, often long-stalked, dark brown (Fig. 9c–e). Conidiogenous cells formed in the swollen base. Ostiole 13–15 μm. Conidia 6.5×5 μm, hyaline, ellipsoidal, 1celled (Fig. 9g). 117 Material examined: France, unlocalised, ex herbarium Léveillé in herbarium Berkeley (as Capnodium citri) [K(M) 172364—iconotype only?, specimen missing]. Ibid., on Olea leaf, ex herbaria C. Montagne, PC, in herbarium Berkeley (as Fumago citri Pers. non Turpin) [K(M) 172363—type?]. Capnodium coartatum Chomnunti & KD Hyde, sp. nov. (Fig. 10a–k) MycoBank 563362 Polychaeton citri (Pers.) Lév. similis sed conidia 4.2–4.6× 1.9–2.4 μm differt. Etymology: from the Latin coartata meaning narrow, in reference to pycnidia. Saprobic on sugary exudates from insects growing on the surface of leaves, fruits, stems and other non plant objects. Thallus of dark brown mycelium growing over the surface of the plant with abundant pycnidia, produced on 3–5 μm wide (x ¼ 4 mm:, n=20), irregularly branched, pale brown to brown, septate, sub-cylindrical hyphal cells, constricted at the septum (Fig. 10f–h). Pycnidia 332–401×34–56 μm (x ¼ 366  45 mm, n=20), superficial, scattered or gregarious, blackish brown, synnematous in structure, ovoid to flask–shaped, elongate, somewhat branched, comprising mostly cylindrical cells, with slightly swollen or flattened base, base of pycnidium forming a pseudo–parenchymatous to prosenchymatous tissue, upper cylindric region tapering to apex, 7.6–11.5 μm wide (x ¼ 9:8 mm, n=10) (Fig. 10a–d), ostiole surrounded by hyaline hyphae (Fig. 10j, k). Conidia 4.2–4.6×1.9–2.4 μm (x ¼ 4:4  2:1 mm, n=20), produced within the swollen base, gathering in a terminal droplet, ellipsoidal, smooth, round ends, hyaline (Fig. 10l, m). Material examined: THAILAND, Chiang Rai Province, Baan Du, on living leaf of Psidium guajava (Myrtaceae), 24 September 2009, SC Karunarathna DPC 040 (MFLU100076, holotype), extype living culture in (MFLUCC100005); Ibid., Mae Fah Luang University, on living leaf of Alstonia scholaris, 3 October 2009, P. Chomnunti DPC 041 (MFLU10-0077), living culture in MFLUCC10-0006. Capnodium tiliae (Fuckel) Sacc.. Syll. fung. (Abellini) 1: 74 (1882) ≡ Capnodaria tiliae (Fuckel) Theiss. & Syd. Ann mycol.15(6): 474 (1918) (1917) ≡ Fumago tiliae Fuckel. Jb. Nassau. Ver. Naturk 23–24: 142 (1870) (1869–70) Saprobic on sugary exudates from insects growing on bark (Fig. 11a). Thallus of superficial mycelium comprising membranous, cylindrical, 4–6 μm (x ¼ 5 mm, n=20) wide, septate hyphae, constricted at the septa, dark brown at the septum and margin (Fig. 11c). Ascomata 81–136 μm diam, 78–141 μm high (x ¼ 96  102 mm, n=10), superficial, globose, without setae, brown to dark brown (Fig. 11b–d, f). Peridium 17–18 μm wide (x ¼ 15 mm, n=15), comprising 118 Fungal Diversity (2011) 51:103–134 Fig. 9 a–g Capnodium citri (type?). a Superficial mycelium on host. b, f Cylindrical, septate mycelium. c–e Elongate pycnidia. g Conidia. Bars: b, c=200 μm, d, e=100 μm, f=50 μm, g=20 μm dark brown cells of textura angularis, darker externally. Hamathecium comprising asci and 2.3–3.6 μm (x ¼ 2:8 mm, n=20) hyaline cells (Fig. 11e). Asci 41–44×15–18 μm (x ¼ 42  17 mm, n=5), 10-spored, bitunicate, clavate, apedicellate, ocular chamber not apparent (Fig. 11g–i). Ascospores 13–17×5–7 μm (x ¼ 15  6 mm, n=20), oblong or ovoid and some reniform, 3–5 septate, constricted at the septa, rarely with longitudinal septa, ends narrowly round, brown, dark brown at septum and margin, wall verrucose (Fig. 11j). Material examined: GERMANY, Biebrich, on branch of Tilia parviflora, Herbier Fuckel no. 899 (1894), (G, Herbier Barbey Boissier). Scorias Fr. Syst. Orb. Veg. 1: 171 (1825) Fungal Diversity (2011) 51:103–134 Fig. 10 a–m Capnodium coartatum (holotype). a–d Stalked pycnidia. e Immature pycnidia. f Mycelia. g, h Pycnidia rising from mycelium. i Abundant conidia at apex pycnidia and pycnidia wall. j, k 119 Ostiole surround by hyaline hyphae. l, m=Conidia. Bars: a–d= 200 μm, f=100 μm, e, g, i–k=50 μm, l, m=20 μm 120 Fungal Diversity (2011) 51:103–134 Fig. 11 a–j Capnodium tiliae (exsicata of Fumago tiliae). a Label data. b Vertical section through ascoma. c Dark brown mycelium. d Peridium. e Peridium with cells of textura angularis. g–i Cylindrical to cylindric-clavate asci. j Ascospores 3–4 septate. Scale bars: f= 200 μm, b–d=100 μm, e, g–i=50 μm, j=20 μm Algorichtera Kuntze, Revis. gen. pl. (Leipzig) 2: 637 (1891) Antennella Theiss. & Syd., Ann mycol. 15(6): 473 (1918) (1917) Antennellina J.M. Mend., in Stevens, Bulletin of the Bernice P. Bishop Museum, Honolulu, Hawaii 19: 55 (1925) Hyalocapnias Bat. & Cif., Saccardoa 2: 114 (1963) Leptocapnodium (G. Arnaud) Cif. & Bat., in Batista & Ciferri, Saccardoa 2: 121 (1963) Paracapnodium Speg., Anal. Mus. nac. B. Aires, Ser. 3 12: 325 (1909) Scolecoxyphium Cif. & Bat., Publções Inst. Micol. Recife 47: 5 (1956) Xystozukalia Theiss., Verh. zool. bot. Ges. Wien 66: 357, 358 (1916) Saprobic on sugary exudates from insects growing on host. Thallus comprising black dense, septate mycelium with ascomata covering the surface of host. Ascomata dark brown to blackish, shiny, subglobose to broadly ellipsoidal, Fungal Diversity (2011) 51:103–134 with a rounded apex, central ostiole present at maturity, with short stalk. Peridium composed of cells of textura angularis. Asci 8-spored, bitunicate, oblong to saccate. Ascospores hyaline, fusiform, with 3–4 trans-septa, the upper cells slightly wider than the lower cells. Pycnidia long stalked, flask-shaped, tapering to the apex, pycnidium wall showing helical twisting, synnemata-like, dark brown to black at the base, brown to pale brown towards the tapering apex. Conidia ellipsoidal, unicellular, hyaline. Anamorphs: Scolecoxyphium Cif. & Bat. (Hyde et al. 2011). Type species Scorias spongiosa (Schwein.) Fr. Syst. mycol. (Lundae) 3 (2): 291 (1832) ≡ Botrytis spongiosa Schwein., Schr. naturf. Ges. Leipzig 1: 127, (1822) Description from type (Fig. 12a–p) Saprobic on sugary exudates from insects growing on host (Fig. 12a). Thallus comprising 3.8–5.5 μm wide (x ¼ 4:7 mm, n=20) wide, black, dense, septate mycelium. A s c o m a t a 7 2 – 8 8 μ m d ia m , 8 9 – 1 3 2 μ m h i g h (x ¼ 82  110 mm, n=5), covering the surface of thallus, gregarious, dark brown to black, shiny, subglobose to broadly ellipsoidal, with a rounded apex, central ostiole present at maturity, on a stalk (Fig. 12c–j). Peridium 14–25 μm (x ¼ 18 mm, n=20), composed of cells of textura angularis (Fig. 12k). Asci 8-spored, bitunicate, oblong to saccate, apedicellate, with a long ocular chamber (Fig. 12l–n). Ascospores 13–15×2–4 μm (x ¼ 13  3 mm, n=20), hyaline, fusiform, 3–4 trans-septate, the upper cells slightly wider than the lower cells (Fig. 12o, p). Description of anamorph from Thai collection (Figs. 13a–g and 14a–k) Saprobic on sugary exudates from insects growing on the surface of living leaves (Fig. 13a). Thallus comprising 3.2– 4.8 μm wide (x ¼ 4 mm, n=20), superficial, cylindrical, septate pale brown hyphae (Fig. 13a). Pycnidia 412–614× 40–57 (x ¼ 503  47 mm, n = 10), long stalked, flaskshaped, tapering to the apex, pycnidial wall showing helical twisting, synnemata-like, frequency with immature ascomata, dark brown to black at the base, brown to pale brown towards the tapering apex (Fig. 13b–f). Conidia 3.1–4.2× 1.6–2.4 (x ¼ 3:7  2 mm, n=20), ellipsoidal, unicellular, hyaline (Fig. 13g). Colonies reaching up to 3 cm diam. after 10 days on PDA medium, flattened, spreading radialy towards the edge, dark-brown in older parts, but olive-green 121 towards the edge. Mycelium 2.6–4.4 μm [x ¼ 3:5 mm, n= 20], consisting of cylindrical and septate hyphae, branched, thick wall, and mucilaginous in the outer layer, with numerous aerial hyphae (Fig. 13a). Pycnidia stalked, flask-shaped, arising from the mycelium plate, comprised of hyaline hyphae helically twisting towards the apex surrounding the ostiole. Stalk pale olive-green and hyaline at the apex, swelling at base (Fig. 13b–g). Conidia 3.5– 4.4 × 1.5–2.2 μm (x ¼ 3:9  1:9 mm, n = 20), ellipsoid, unicellular, guttulate, hyaline (Fig. 14i–k). Material examined: Type specimen: ‘Car. N. 1311. Podiosoma? epiphega. Schwein.! In litt.’ (UPS Fries— lectotype). USA?, unlocalised, ex herbarium Schwein., in herbarium M.J. Berkeley [K(M) 171138—syntype]; ibid. Ohio, on wood, March, H.W. Ravenel 145 (as Scorias spongiosa) [K(M) 171139 & ? IMI 30376—authentic]; ibid. South Carolina, ad ramos Alni, Dec. H.W. Ravenel 1384 (as Scorias spongiosa) [K(M) 171140—authentic]; ibid. Ohio, on Fagus sp., ex herbarium M.J. Berkeley (as Scorias spongiosa) [K(M) 171141—authentic]; ibid. Ohio, on leaf and bark of Fagus sp., ex herbarium M.J. Berkeley (as Scorias spongiosa) [K(M) 171142—authentic]; THAILAND, Chiang Rai Province, Khunkorn, on living leaf of Entada sp. (Fabaceae), 18 December 2009, Putarak Chomnunti, DPC048 (MFLU10-0013, epitype designed here), living culture in MFLUCC10-0084. The species has been sanctioned by Fries Fr., Syst. mycol. 3(2): 291 (1832) with a collection from Carolina number 1311. In herbarium M.J. Berkeley at K there are five collections labeled Scorias spongiosa from Ohio numbered 145 found on Fagus leaves and twigs, and one from South Carolina collected by Ravenel on Alnus without a number, and a representation of these have been examined by Reynolds (1975). All of those are authentic material of Scorias spongiosa according to Saccardo (Sylloge Fungorum I: 83, 1882). Nevertheless, there is a further collection in herbarium Berkeley, originally from herbarium Schweinitz, which contains no further annotations, but maybe part of Fries’s listed 1,311 specimen. From this collection we have chosen to make a slide, and the description is included above. Further to the above the IMI 230376 herbarium (now part of K herbarium’s holdings) contains several slides labeled type from Berkeley’s material, but it is not clear what specimen they used for preparing the collection. The type illustrated contained only the teleomorph. The fresh collection from Thailand contained only the mature anamorph and immature ascomata. In the molecular analysis Scorias spongiosa (CBS 325.33) clustered with strain MFLU10-0084 with 100% bootstrap support and is clearly placed in Capnodiaceae. Nine species are currently recorded in the genus in Index Fungorum (http://www.indexfungorum. org/Names/Names.asp; accessed on 12 July 2011). 122 Fungal Diversity (2011) 51:103–134 Fungal Diversity (2011) 51:103–134 a–p Scorias spongiosa (syntype). a, b Ascomata on hyphal mass on host. c, d Squash of ascomata. e–j Vertical section through ascomata on hyphal mass. k Stalked ascomata. l–n Asci with thick wall. f, g Pale brown ascospores. Bars: d–h, j=100 μm, c, i=50 μm, k–n=20 μm, o, p=10 μm ƒFig. 12 Genera of Capnodicaceae incertae cedis Scoriadopsis J.M. Mend., in Stevens, Ann mycol. 28(5/6): 365 (1930) Scoriadopsis miconiae J.M. Mend., in Stevens, Ann mycol 28(5/6): 365 (1930) ≡ Rizalia miconiae (J.M. Mend.) E. Müll., in Müller & von Arx, Beitr. Kryptfl. Schweiz 11 (no. 2): 597 (1962) Colonies sooty black, on upper surface and closely associated with Meliola. Thallus perisporioid, slimy, loosely woven, weft-like, hyaline to straw-colored hyphae. Ascomata globose or ovoid, ostiolate, stalked, gelatinous, dark brown. Asci 40× 14 μm, numerous, 8-spored, ovate, pseduoparaphyses lacking. Ascospores 16×4 μm, fusiform, 1-septate (Stevens 1930). Fig. 13 a–g Scorias spongiosa (epitype). a Ascomata and pycnidia on surface of leaf. b, d–f Immature ascomata and pycnidia. c Pycnidia arising from mycelium. g Conida. Bars a=200 μm, d, e=100 μm, b, f=50 μm, g=20 μm 123 We have not seen type material of this species. The taxon is similar to Scorias, but is has 1-septate ascospores, while Scorias has 3-4 trans-septate ascospores which are pigmented when mature. We retain Scoriadopsis in Capnodiaceae incertae sedis as colonies appear like sooty moulds on the host and ascomata are globose, black, and ascospores turn brown when mature. However, the melioaceous habit would be unusual. Genus transferred to Mycosphaerellaceae Echinothecium Zopf, Nova Acta Acad. Caes. Leop.— Carol. German. Nat. Cur. 70: 250 (1898) Thallus superficial, composed of thick brown often anastomosing hyphae. Ascomata globose, ostiolate, provided with stiff, simple, appendages. Asci 8-spored, sessile, lacking paraphyses. Ascospores 2 celled, hyaline, oval, the upper cell slightly wider than the lower cell. Anamorphs: None. 124 Fungal Diversity (2011) 51:103–134 Fig. 14 a–k Scorias spongiosa (epitype). a Hyphae. b Immature pycnidia. c Mycelium bearing pycnidium. d–g Mature pycnidia. h Septate hyphae. i–k Hyaline conidia. Bars: a=200 μm, e–g=100 μm, b–d=50 μm, h–j=20 μm, k=10 μm Type species Sphaerellothecium reticulatum (Zopf) Etayo, Cryptog. Mycol. 29(1): 87 (2008) ≡ Echinothecium reticulatum Zopf, Nova Acta Acad. Caes. Leop.—Carol. German. Nat. Cur. 70: 250 (1898) (Fig. 15a–l) Thallus comprising 4–6 μm thick (x ¼ 5 mm, n=20), superficial, dense, brown, often anstomosing, reticulate, branching, septate hyphae (Fig. 15a, f, g). Ascomata 32– 68 μm diam, 31–69 μm high (x ¼ 50  49 mm, n=5), spherical or depressed-globular, dark brown, surrounded by 14–18×4–6 μm (x ¼ 16  5 mm, n=10) hyphal appendages (Fig. 15b–e). Asci 23–36×10–13 μm (x ¼ 27  11 mm, n= 10), 8-spored, bitunicate, ovoid, apedicellate, with an ocular chamber (Fig. 15h–j). Ascospores 8–6 × 2.5–4 μm (x ¼ 7  3 mm, n = 20), ovoid, hyaline, the upper cell slightly wider than the lower cell, 1-septate, constricted at the septum (Fig. 15k, l). Material examined: ITALY, South Tyrol, Fondo, Mendehof, on bark of spruce trees on Parmelia sulcata, 18 August 1897, Arnold exs. 1743 (K(M) 171135, 171136 syntypes!). Fungal Diversity (2011) 51:103–134 125 Fig. 15 a–l Sphaerellothecium reticulatum (syntype of Echinothecium reticulatum). a Ascomata on Parmelia sulcata. b–e Ascoma with eternal hyphal appendages. f, g Mature hyphae. h–j Bitunicate asci. k, l Ascospores. Bars: a, c, d, f=50 μm, b, e, g–i=20 μm, j, k=10 μm Echinothecium occurs on lichens and has been found in Europe and North America (Navarro-Rosines and GomezBolea 1989). Lichenostigma cosmopolites Hafellner & Calat has been most wrongly identified and confused with Echinothecium reticulatum which has ascomata with septate seta-like hyphae and seems to be restricted to Parmelia s. str. (Calatayud et al. 2002). This is the only genus in the Capnodiaceae which has a lichenicolous habit. Etayo (2008) placed this species in the genus Sphaerellothecium and after critical study of the holotype of Sphaerellothecium we agree with his decision. We have examined the holotype of Sphaerellothecium and it is similar to Echinothecium and thus considered a synonym. The generic type, Sphaerellothecium araneosum is described below. Sphaerellothecium araneosum (Rehm) Zopf, Nova Acta Acad. Caes. Leop.—Carol. German. Nat. Cur.70: 178 (1897) (Fig. 16a–l) ≡ Sphaerella araneosa Rehm, Ascomyceten Dign.: no. 133 (1872) ≡ Discothecium araneosum (Rehm) Vouaux, Bull. Soc. mycol. Fr. 29: 55 (1913) = Echinothecium glabrum M.S. Christ., Alstrup & D. Hawksw., in Alstrup & Hawksworth, Meddr Grønland, Biosc. 31: 28 (1990) ≡ Endococcus araneosus (Rehm) H. Olivier, Bull. Acad. Intern. Géogr. Bot. 17: 127 (1907) ≡ Epicymatia araneosa (Rehm) Sacc., Syll. fung. (Abellini) 1: 572 (1882) 126 Fungal Diversity (2011) 51:103–134 Fig. 16 a–k Sphaerellothecium araneosum (holotype of Sphaerella araneosa). a Fruiting bodies on host tissue. b Ascomata with hyphae. c Section through ascoma. d Section through peridium. e–g Ascus. h–j Ascospores. k Mature ascospore. Bars: a=200 μm, b=100 μm, c=50 μm, d=20 μm, e–k=10 μm ≡ Mycosphaerella araneosa (Rehm) Lindau, Hilfsb. Sammeln Ascomyc. (Berlin): 125 (1903) ≡ Phaeosphaerella araneosa (Rehm) Sacc. & D. Sacc., Syll. fung. (Abellini) 17: 676 (1905) Thallus with superficial net of dark brown, septate, vegetative hyphae on host tissue (Fig. 16a). Ascomata 43–72 μm diam, (42–) 45–79 μm high (x ¼ 57:4  57:3 mm, n=10), superficial on host surface, visible as black dots, uniloculate, individually globose to subglobose, scattered or gregarious, dark to dark brown (Fig. 16a–c). Peridium 3–6 (−9.5) μm wide (x ¼ 4:6 mm, n=10), comprising 1–2 layers of dark to dark brown cells of textura angularis (Fig. 16d). Asci (26–) 30–37 (−45)×16–21.5(−25) μm (x ¼ 35  19 mm, n=10), 8-spored, bitunicate, saccate, ovoid or occasionally obclavate, non pedicellate, apically rounded with an ocular chamber (Fig. 16e–g). Ascospores (11–)13–17.5×6–8.5 (−9.5) μm (x ¼ 14:7  6:8 mm, n=20), irregularly seriate, Fungal Diversity (2011) 51:103–134 fusiform to ellipsoidal, both ends obtuse, 1-septate, constricted at the central septum, upper cell larger than lower cell, thick-walled, initially hyaline, becoming brown when mature (Fig. 16h–k). Material examined: AUSTRIA, Tyrol, Materi am Brenner, Waldrast, on the thallus and apothecia of Ochrolechia pallescens var. upsaliensis (L.), Alt. 6700’, August 1872, F. Arnold (M-0044221, holotype). Genera transferred to Chaetothyriaceae The following genera do not belong is Capnodiaceae and are placed in Chaetothyriaceae. Aithaloderma Syd. & P. Syd. Ann mycol. 11: 256 (1913) Blastocapnias Cif. & Bat., Saccardoa 2: 67 (1963) Chaetopotius Bat., Mycopath. Mycol. appl. 5: 151 (1951) Ciferrioxyphium Bat. & H. Maia, in Batista & Ciferri, Quad. Lab. crittogam., Pavia 31:65 (1963) Hypocapnodium Speg., Physis, B. Aires 4: 287 (1918) Phaeochaetia Bat. & Cif., Beih. Sydowia 3: 62 (1962) Vertixore V.A.M. Mill. & Bonar, University of Calif. Publ. Bot. 19: 406 (1941) Saprobic on sugary exudates from insects growing on leaves of various plants. Thallus comprising superficial, brown to black, septate hyphae, constricted at the septa, mostly dense and anastomosing, covering the surface of leaves, forming a dark mycelia mat. Ascostromata superficial, brown to dark brown, in vertical section globose to subglobose, covered by subiculum or ascosomatal layer, consisting of brown, septate hyphae, with a circumferential space around the maturing ascomata, which results from expanding the expansion of the ascomata. Towards the upper part of the ascomata wall, the pellicles are very tightly packed, with 5–10 short black and dull setae which surround the ostiole. Peridium comprising cells forming a textura angularis, pale brown to hyaline; without interthecial filaments. Asci 8-spored, bitunicate, wide fusiform to saccate, short pedicellate, lacking an ocular chamber. Ascospores hyaline, trans-septate, some surrounded by a mucilaginous sheath. Aithaloderma was assigned to Capnodiaceae by Sydow and Sydow (1913) and currently includes 15 species (Kirk et al. 2008). Aithaloderma was placed in Chaetothyriaceae by Spegazzini (1918) based on the dome-shaped or flattened ascostromata and pleomorphic pycnidia. Hughes (1976) suggested that Aithaloderma could be placed within Capnodiaceae because the mycelial hyphae are similar to those in Phragmocapnias. Recently, Aithaloderma ferrugineum Fraser has been reviewed by Reynolds and Gilbert (2005), who also included the species within the Capnodiaceae pending molecular data. The genus is characterized 127 by globose perithecium with setae, lageniform pycnidia which produce continuous, hyaline conidia and funnel to funnel-globulose ascostromata with short setae especially around the ostiole, lack paraphyses and asci contain 8-spored, transversely septate, clavate, hyaline ascospores (Yamamoto 1954; von Arx and Müller 1975; Hughes 1976). Leptoxyphium was reported to be the anamorph of Aithaloderma but in this study Leptoxyphium clusters in Capnodiaceae. Since there is no reported connection between Leptoxyphium and Aithaloderma clavatisporum (the generic type) we transfer Aithaloderma to Chaetothryriaceae while considering Leptoxyphium within Capnodiaceae. Anamorphs: ?Ciferrioxyphium Bat. & H. Maia, Leptoxyphium Speg. (Hyde et al. 2011). Type species Aithaloderma clavatisporum Syd. & P. Syd. Annls. Mycol. 11: 256 (1913) (Fig. 17a–m) ≡ Chaetothyrium clavatisporum (Syd. & P. Syd.) Hansf., Mycol. Pap. 15 (1946) ≡ Phaeochaetia clavatispora (Syd. & P. Syd.) Bat. & Cif. Beih. Sydowia 3: 67 (1962) Saprobic on sugary exudates from insects growing on the surface of living leaves (Fig. 17a) Thallus comprising 2.4− 4.1 μm thick (x ¼ 3:5 mm, n=20), superficial, brown to black, septate, hyphae, constricted at the septa, mostly with 90° branching, mostly dense and anastomosing, covering the surface of leaves forming a dark mycelia mat (Fig. 17b, f). Ascostromata 100 − 124 μm diam, 80 − 122 μm high (x ¼ 122  100 mm, n=5), superficial, brown to dark brown, initials arising from cells of mycelium, in vertical section globose to subglobose, covered by a subiculum or a layer of brown septate hyphae, with a circumferential space around the maturing ascomata, which results from the expansion of the ascomata. Towards the upper part of the ascomata wall, the pellicles are very tightly packed, with 5–10 short black and dull setae surrounding the ostiole (Fig. 17b–e). Peridium 4.51–9 μm wide (x ¼ 7 mm, n=20), a single layer, comprising of cells forming a textura angularis, pale brown to hyaline. Pseudoparaphyses not observed. Asci 39–69×10– 31 μm (x ¼ 51  21 mm, n=10), 8-spored, bitunicate, wide fusiform to saccate, short pedicellate, lacking an ocular chamber (Fig. 17h–j). Ascospores 12–25 × 3 − 6 μm (x ¼ 19  4:5 mm, n=20), overlapping triseriate, fusiform, hyaline, 4–5 septate, constricted at the septa, upper cells wider than lower cells, some surrounded by a mucilaginous sheath (Fig. 17k–m). Material examined: PHILIPPINES, Laguna Province, Los Baños, in living? leaves of Voacanga globosa (Apocynaceae), 1 February 1913, CF. Baker, Sydow, Fungi Exotici Exsiccata 174 (IMI 26048, isotype). 128 Fungal Diversity (2011) 51:103–134 Fig. 17 a–l Aithaloderma clavatisporum (isotype). a Sooty mold and ascostromata on surface of host. b Ascostromata with short setae. c–e Section through ascostromata. f Network of hyphae. g–i Asci. j–l Ascospores. Bars: b–d=100 μm, e–g=50 μm, h–l=20 μm Some morphological characters of Aithaloderma are similar to those found in Chaetothyriaceae (Batista and Ciferri 1962; Chomnunti et al. 2012). These include the nature of the superficial ascostromata on a mycelial mat with a circumferential space around the subglobose maturing ascomata, with setae surrounding the ostiole, wide fusiform to saccate asci and clavate, hyaline, 5septate ascospores. These characters indicate that Aithaloderma is better placed in Chaetothyriaceae and we follow this here. Ceramoclasteropsis coumae Bat. & Cavalc., in Batista, Perez & Bezerra, Brotéria, sér. Ci. Nat. 31(2): 101 (1962) This species has superficial ascomata that form on superficial dark brown mycelium and appear to be ascostromata (Batista et al. 1962). Asci are bitunicate and apedicellate and ascospores are 7.5–14×2–4 μm, clavate, hyaline with 1–3 transverse septa. The taxon is reported to have paraphyses as in typical members of the Chaetothyriaceae. The ascospores appear to be typical of Limacinula and fresh collections are required to confirm if this is a distinct genus. Ceramoclasteropsis Bat. & Cavalc., in Batista, Perez & Bezerra, Brotéria, sér. Ci. Nat. 31(2): 101 (1962) Hyaloscolecostroma Bat. & J. Oliveira, Atas Inst. Micol. Univ. Pernambuco 5: 448 (1967) Fungal Diversity (2011) 51:103–134 Hyaloscolecostroma rondoniense Bat. & J. Oliveira, Atas Inst. Micol. Univ. Recife 5: 449 (1967) This genus was described from Brazil and is monotypic. This is a sooty mould with superficial ascomata, cylindrical unitunicate asci and trans-septate cylindric-fusiform ascospores (Batista and da Silva 1967). Apparently this fungus is associated with homopterean insects. The drawing provided in Batista and da Silva (1967) this is not a convincing taxon of Capnodiaceae especially as it shows a thin unitunicate ascus layer. Thus we place the genus in the Chaetothyriaceae incertae cedis. Trichomerium Speg., Physis, B. Aires 4: 284 (1918) Trichomerium coffeicola (Puttemans) Speg., Physis, B. Aires 4: 284 (1918) ≡ Limacinia coffeicola Puttemans, Cryptog. Mycol. 20: 163 (1904) Trichomerium is included in Capnodiaceae in Index Fungorum (http://www.indexfungorum.org/Names/Names. asp), but not in Lumbsch and Huhndorf (2010). The genus was introduced by Spegazzini (1918) with type species T. coffeicola (Reynolds 1982) and include 23 species, mostly folicolous fungi. The ascospores are similar to Phragmocapnias and Scorias in septation and in being hyaline, but Trichomerium has sessile ascomata. We have made several collections of Trichomerium in this study which cluster in Chaetothyriaceae in the phylogenetic analysis (data not shown) and will be reported in a later paper. Anamorphs reported for genus: Tripospermum Speg. (Hyde et al. 2011). Tripospermum Speg., Physis, B. Aires 4(no. 17): 295 (1918) = Pentaposporium Bat., Revta Biol., Lisb. 1(2): 106 (1957) Tripospermum acerinum P. Syd., Physis, B. Aires 4(no. 17): 295 (1918) Tripospermum was introduced by Corda in 1837 for T. elegans and by Spegazzini in 1918 as segregate of Tripospermum with type species T. acerinum (Hughes 1951). As the anamorph of Trichomerium Speg. (Hyde et al. 2011) it should be transferred to Chaetothyriaceae. Genus in Coccodiniaceae Microxiphium (Harv. ex Berk. & Desm.) Thüm., Physis, B. Aires 4(17): 293 (1879) Microxiphium footii (Harv. ex Berk. & Desm.) Thüm., Physis, B. Aires 4(17): 293 (1879) This genus is polyphyletic with species in Coccodiniaceae and Capnodiaceae. However the type of the genus is presently placed as synonym of Dennisiella babingtonii and thus included in the Coccodiniaceae. The specimen of Microxyphium citri used in the phylogenitcal tree (see 129 Fig. 1) is likely to be a misidentification. Microxyphium citri, M. aciculiform and M. theae are also dispersed amongst the Capnodiaceae in the tree indicating its polyphyletic nature. Genus transferred to Micropeltidaceae Wu et al. (2011) discusses this family and the following genera are better placed therein. Callebaea Bat. in Batista, Perez & Bezerra, Brotéria, sér. bot. 31: 100 (1962) Mycelium superficial, irregularly scattered. Thyriothecia circular, scattered, superficial, membraneous, brownish, lower peridium poorly developed easily removed from the host surface, with a central irregular ostiole; in section lenticular. Upper wall comprising an irregular meandering arrangement of compact hyphae. Hamathecium comprising asci, pseudoparaphyses not obvious. Asci immature and ascospores not observed. Anamorphs: None. Type species Callebaea rutideae (Hansf.) Bat., In Batista, Perez & Bezerra, Brotéria, sér. bot. 31: 100 (1962) (Fig. 18a–k) ≡ Microcallis rutideae Hansf., Proc. Linn. Soc. London 158: 40 (1947) ≡ Microcalliopsis rutideae (Hansf.) Bat. & Cif., Bein. Sydowia 3: 58 (1962) Foliar epiphyte on upper surface of leaves, lacking superficial mycelium (Fig. 18a, b). Thyriothecia 83 (−196)–228 μm diam × 41–63(−85) μm high (x ¼ 183  59 mm, n=5), circular, scattered, superficial, pale brown to brown, basal peridium poorly developed, easily removable from the surface of leaves, lacking an ostiole; in section lenticular (Fig. 18e). Upper wall comprising an irregular meandering arrangement of compact hyphae, 2–3 μm wide (x ¼ 2:5 mm, n = 10) (Fig. 18c, d). Hamathecium comprising asci, pseudoparaphyses not obvious. Asci immature (Fig. 18f–k) and ascospores not observed. Material examined: UGANDA, Entebbe Road, in leaves of Rutideae smithii (Rubiaceae), August 1944, C. G. Hansford 3560 (K(M) 164029, holotype). Hansford (1947) introduced Microcallis rutidaea as new species in Chaetothyriaceae. We examined type material and found a few flattened ascomata on host surface and these lacked mature asci and ascospores. The thyriothecium however, indicate that Microcallis rutideae should be transferred to Micropeltidaceae. However the specimen is immature and must be treated as doubtful until a better specimen or fresh collections are found. 130 Fungal Diversity (2011) 51:103–134 Fig. 18 a–k Microcallis rutideae (holotype). a, b Ascomata on leaf surface. c, d Circular ascomata comprising meandering hyphae. e Vertical section of ascostomata. f–k Young asci. Bars: c, e=100 μm, d, j=50 μm, f–i, k=20 μm Doubtful genera Type species Anopeltis Bat. & Peres. Nova Hedwigia 2: 472 (1960) Foliar epiphyte on leaves. Thallus comprising superficial, olive-brown, cylindrical, branching, irregularly reticulate, septate hyphae, lacking setae. Ascostromata semi-immersed, dark-brown to brown at the margin, flattened, round, scattered, surrounded with mycelium, the locule/asci produced under the stromata, hypostromata central or lateral. Ascostromata wall composed of angular brown cells. Peridium thin. Conidia clavate or oblong, 1-septate, deeply constricted at the septum, thick walled, wrinkled. Anamorphs: None. Anopeltis venezuelensis Bat. & Peres, Nova Hedwigia 2: 472 (1960) (Fig. 19a–j) Foliar epiphyte on leaves (Fig. 19a). Thallus superficial, 2.7–5 μm wide (x ¼ 3:7 mm, n=20), comprising olivebrown, slender, branched, septate, cylindrical hyphae, irregularly reticulate, lacking setae. (Fig. 19b). Ascostromata 145–208 μm diam, 74–93 μm high (x ¼ 175  84, n=5), uniloculate, scattered, semi-immersed, dark-brown to brown, surrounded by mycelium. The wall of ascostromata composed of angular brown cells (Fig. 19e, f). Peridium thin, 2–3 μm wide (x ¼ 2:5 mm, n=10) (Fig. 19d, g). Dark brown conidia cover and surround the ascostromata. Fungal Diversity (2011) 51:103–134 131 Fig. 19 a–j Anopeltis venezuelensis (holotype). a Appearance of leaves colonized by the fungus. b Ascostromata with mycelium and conidia. c Pale brown angular cells of ascostromata edge. d Section of ascostomata with angular cells. e Young locule/asci. f Vertical section of ascostromata with young asci. g Angular cells of ascostromata. h–j Brown conidia with 1-septa. Bars: b–d=100 μm, c, e–g=20 μm, h–j=10 μm Conidia 9–14×3–6 μm (x ¼ 11  4, n=15), clavate or oblong, 1-septate, deeply constricted at the septum, thick walled and wrinkled (Fig. 19h–j). Material examined: VENEZUELA, Miranda, Caracas, Los Palos Grandes (above), Mt Naiguata (S slope of), c. 1,200 m, in the leaves of unidentified plant, 13 July 1959, R.W.G. Dennis 2321 (K(M) 171577—holotype). Anopeltis venezuelensis should be placed in Ascomycete incertae cedis according to its semi-immersed ascomata, lack of mycelium and peridium of textura angularis. The nature of this taxon is not clear from the type specimen as asci or ascospores were not observed. The taxon needs to be recollected and examined from fresh material. Capnophaeum Speg. Physis, B. Aires 4: 287 (1918) Capnophaeum indicum C. Bernard, Physis, Rev. Soc. Arg. Cienc. Nat. 4: 287 (1918) We requested the type specimen from BO but was informed it is lost and therefore the genus must be considered as doubtful. Fumiglobus D.R. Reynolds & G.S. Gilbert., Cryptog. Mycol. 27(3): 252 (2006) Fumiglobus ficina (Bat., Nascim. & Cif.) D.R. Reynolds & G.S. Gilbert., Cryptog. Mycol. 27(3): 253 (2006) ≡ Asbolisia ficina Bat. Nascim. & Cif. in Batista & Ciferri, Quad. Lab. crittogam. Pavia 31: 41 (1963) 132 Fumiglobus was introduced by Reynolds and Gilbert (2006) as new genus to accommodate Asbolisia nomen confusum, which included nine species; typified by somewhat aerial, membraceous, superficial mycelium, comprised of dark brown septate hyphae, constricted at the septum, with pycnidia borne from several hyphae. Conidia are globose to subglobose, hyaline, single-celled, bacillate to ellipsoidal, or cylindrical. Fresh collections are needed to establish if the genus is distinct. Polychaetella Speg., Physis, B. Aires 4: 295 (1918) Polychaetella schweinitzii (Berk. & Desm.) Speg., Physis, B. Aires 4: 295 (1918) ≡ Capnodium schweinitzii Berk. & Desm. (1849) Polychaetella was classified in Capnodium section IV by Saccardo (1822) and were anamorphs of various Capnodium species (Sutton 1977), and have elongated pycnidia producing dictyospetate, hyaline or chlorinous conidia (Hughes 1976). Batista and Ciferri (1963a, b) added Polychaetella in Asbolisiaceae and accepted P. araucariaceae (Thüm) Speg. as type species. Hughes (1976) observed the DAOM syntype of Capnodium araucariae but could not find the hyaline dictyospetate conidia. Therefore the genus must be treated as uncertain. Discussion We examined nine generic types and sequenced 15 taxa of Capnodiaceae. Phylogenetic analysis show Capnodium, Leptoxyphium, Phragmocapnias and Scorias to be well defined genera in Capnodiaceae, while Aithaloderma, Anopeltis, Callebaea, Echinothecium and Trichomerium are removed to other families or are doubtful. This study thus provides an expansion in the documentation of capnodiaceous sooty molds. This group now comprise three easily recognized genera, i.e. Phragmocapnias = Conidiocarpon, Capnodium = Polychateon and Scorias, comprising of both sexual and asexual states, and the exclusively anamorphic Leptoxyphium. It is not clear whether the genera Anopeltis, Capnophaeum and Scoriadopsis however, are acceptable in Capnodiaceae yet. The present study has shown that the general term “sooty molds” encompasses a broad set of species within the families Antennulariellaceae, Capnodiaceae, Chaetothyriaceae and Metacapnodiaceae (Hughes 1976; Reynolds 1986, 1998; Chomnunti et al. 2012) which often grow in close association on a single leaf. This had posed considerable challenges to to taxonomists since the mid-19th century (Friend 1965; Hughes 1976; Reynolds 1986, 1998; Chomnunti et al. 2012). Pycnidiaceous sooty moulds were placed in the generally unaccepted family Asbolisiaceae (Batista 1963) and Hosagoudar and Riju (2011) recently Fungal Diversity (2011) 51:103–134 treated them in a new family Schifferulaceae, as black mildews, which he termed “saprobic sooty moulds”. The Schifferulaceae may well be synomymous with Asterinaceae but this cannot be corroborated without DNA sequence comparions. DNA sequence data remains rare in other families too. Only one putative species of Antennulariellaceae (Antennariella placitae) has however, been sequenced and clustered in Capnodiaceae (Cheewangkoon et al. 2009). This study and that of Chomnunti et al. (2012) clearly shows that Capnodiaceae and Chaetothyriaceae consist of unrelated taxa belonging to different classes and yet they can hardly be distinguished by morphology and growth habit. It seems clear that the similar morphological characters in these groups evolved under selection pressures unique to their shared niche to utilise sugary insect exudates on leave surfaces. Besides the species associated with leaf surfaces and particularly with honey dew produced by insects, our analysis indicates other possible environmental niches in Capnodiaceae. One sequence in our analysis was obtained from a member of an ecological guild of rock-inhabiting fungi (Ruibal et al. 2009). Members of this found interspersed throughout the classes Dothideomycetes and Eurotiomycetes, can tolerate extreme conditions found on rock surfaces. It would be interesting to establish if capnodiaceous species grew on proximity to honey dew on rocks surfaces. Another group of sequences were isolated from a species found in ant nest walls. This undescribed species act as mutualists by aiding in reinforcing the nest walls. They are cultivated by the ants which nourishes them with honeydew (Schlick-Steiner et al. 2008). Finally, there are several pathogenic species which occur on Capnodiaceae and include species in the genera Rhombostilbella (Pohlad 1988). It will be important that these taxa are considered carefully when studying sooty moulds to avoid confusion in future. Acknowledgments This work was supported by the Thailand Research Fund BRG528002. The Global Research Network for Fungal Biology and King Saud University are also thanked for support. 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