742 Mycol. Res. 104 (6) : 742–754 (June 2000). Printed in the United Kingdom. Eight new species of Anthostomella from South Africa Bing-Sheng LU, Kevin D. HYDE and Edward C. Y. LIEW Fungal Diversity Research Project, Department of Ecology and Biodiversity, The University of Hong Kong, Pokfulam Road, Hong Kong. E-mail : blua!hkusua.hku.hk Received 27 October 1998 ; accepted 30 August 1999. Eight new species of Anthostomella, A. acuminata, A. applanata, A. caffrariae, A. colligata, A. meerensis, A. palmae, A. raphiae and A. spiralis, are described from South Africa. They are compared with similar species and illustrated with differential interference contrast photomicrographs. INTRODUCTION There are only two species of Anthostomella known from South Africa, A. capensis (Doidge 1948) from Phoenix dactylifera and A. rubicola (Francis 1975). Martin (1969) described three Anthostomella species from South Africa, A. alpigena from Acacia cyclops and Sideroxylon inerme, A. xylostei and A. melanotes from Canthium spinosum and unidentified wood, but A. alpigena and A. xylostei are certainly Amphisphaerella species (Lu, pers. obs.). We have not examined specimens of A. melanotes collected by Martin in South Africa. We can find no other records of species of the genus in South Africa. Collections of Anthostomella on Jujubiopsis sp., Phoenix sp. and Raphia sp. in South Africa were made by K. D. H. and T. Steinke in November 1994. Based on these collections, eight new species are described and illustrated. MATERIALS AND METHODS All specimens examined were rehydrated from dried material and are deposited at HKU(M). Slides of ascospores, asci and sections of ascomata were mounted with distilled water for observation, photomicrography and measurement. Ascal apical rings were stained using Melzer’s solution. Sections of ascomata were made on a Cryotome and mounted with O.C.T. Compound (BDH Laboratory Supplies, England). Francis (1975) attempted to culture the Anthostomella species she examined using different media under different conditions. We have also attempted to culture these species of Anthostomella but without success. TAXONOMY Anthostomella acuminata B. S. Lu & K. D. Hyde, sp. nov. (Figs 1–7, 72) Etym. : from the Latin acuminata meaning ‘ pointed ’, in reference to the ascospores with tapering ends. Ascomata 300–440 µm diam, 280–430 µm alta, globosa, ostiolata, clypeata. Asci 97–125¬6±3–7±5 µm, apparatu subapicali iodo coerulescente, 2±5–3±1 µm diam, 1–1±9 µm alto praediti. Ascosporae 12–14±5¬4±5–5¬3±5–4 µm, ellipsoideae, utrinque acuminatae, fissura dispansa praeditae. Ascomata immersed under a clypeus, visible as blackened, conical areas, 0±17–0±28¬0±17–0±31 mm (xa ¯ 0±21¬ 0±22 mm), clustered or mostly solitary (Fig. 1) ; in vertical section 300–440 µm (xa ¯ 347±5 µm, n ¯ 11) diam, 280– 430 µm (xa ¯ 339±2 µm, n ¯ 11) high, dark brown, coriaceous, globose, with a central periphysate ostiolar canal, 15–40 µm (xa ¯ 23±4 µm, n ¯ 5) diam, 80–110 µm (xa ¯ 87±8 µm, n ¯ 5) high (Fig. 2). Clypeus black, 160–250 µm (xa ¯ 215 µm, n ¯ 10) diam, 60–100 µm (xa ¯ 82±5 µm, n ¯ 10) high around the neck, comprising brown host cells mixed with darkened intracellular fungal hyphae (Fig. 2). Peridium 10–13±8 µm (xa ¯ 11±9 µm, n ¯ 10) wide, comprising several layers of hyaline to light coloured, compressed cells inwardly and darkened, thickwalled, angular cells towards the outside (Fig. 3). Paraphyses 2±5–3±8 µm (xa ¯ 3±1 µm, n ¯ 15) wide, hypha-like, flexuose, septate, numerous and embedded in a gelatinous matrix. Asci 97–125¬6±3–7±5 µm (xa ¯ 110±7¬7 µm, n ¯ 20), eightspored, cylindrical, short-pedicellate, unitunicate, apically rounded, with a weakly J­, wedge-shaped, subapical ring, 2±5–3±1 µm (xa ¯ 2±9 µm, n ¯ 15) diam, 1–1±9 µm (xa ¯ 1±6 µm, n ¯ 15) high (Figs 4–5). Ascospores 12–14±5¬4±5–5¬ 3±5–4 µm (xa ¯ 12±3¬4±8¬3±8 µm, n ¯ 25), uniseriate, ellipsoidal with tapering ends, brown, unicellular, smoothwalled, with hyaline, end caps of mucilage, 2±5–3±1 µm (xa ¯ 2±7 µm) long, 0±6–1 µm (xa ¯ 0±8 µm) wide, germ slit straight, extending over the full length (Figs 6–7). Material examined : South Africa : Richards Bay, Meerens, on the stem of Jujubiopsis sp., 15 Nov. 1994, K. D. Hyde & T. Steinke (HKU(M) 2118–holotypus) ; on stem of Phoenix reclinata, 15 Nov. 1994, K. D. Hyde & T. Steinke (HKU(M) 2117). B.-S. Lu, K. D. Hyde and C. Y. Liew 743 Other material examined : USA : New York, Lyndonvelle, on decorticated branch on ground, May 1889, C. E. Fairman, 42, (NY– holotype of A. eructans.).–Malaysia : Pasoh Forest Reserve, on rachis of Licuala sp., Oct. 1991, K. D. Hyde, 1562a (BRIP 21950–holotype of A. licualicola). Anthostomella acuminata is closest to A. eructans and A. licualicola, but these do not have J­ subapical rings in their asci, A. eructans has non-acuminate ascospores, and those of A. licualicola are inequilaterally broadly ellipsoid-fusiform. Anthostomella applanata B. S. Lu & K. D. Hyde, sp. nov. (Figs 8–15, 74) Etym. : from the Latin applanata meaning ‘ flattened ’, in reference to the flattened ascospores. Ascomata 420–650 µm diam, 250–480 µm alta, subglobosa vel globosa, immersa. Asci 75–118¬7±5–8±8 µm, apparatu subapicali cuneato iodo coerulescente, 3±1–3±8 µm diam, 1–1±5 µm alto praediti. Ascosporae 12±5–16±5¬5±5–6±5¬4–5 µm, inaequilateraliter ellipsoideae, unilateraliter applanatae, appendice mucilaginoso praeditae. Ascomata immersed, visible as tiny blackened papillae, solitary or clustered (Fig. 8) ; in vertical section 420–650 µm (xa ¯ 504 µm, n ¯ 10) diam, 250–480 µm (xa ¯ 341 µm, n ¯ 10) high, dark brown, coriaceous, subglobose to globose, periphysate ostiolar canal, 20–50 µm (xa ¯ 32±5 µm, n ¯ 6) diam, 60–120 µm (xa ¯ 98±3 µm, n ¯ 6) high (Fig. 9). Peridium 3±8–6±3 µm (xa ¯ 4±6 µm, n ¯ 10) wide, comprising several layers of compressed cells, walls hyaline at the inside and brown towards the outside (Fig. 10). Paraphyses 1±3–1±9 µm (xa ¯ 1±5 µm, n ¯ 10) wide, hypha-like, septate, flexuose, numerous and embedded in a gelatinous matrix. Asci 75–118¬7±5–8±8 µm (xa ¯ 97±4¬7±8 µm, n ¯ 20), eightspored, cylindrical, long-pedicellate, pedicel 22±5–26±3 µm (xa ¯ 24±3 µm, n ¯ 10), unitunicate, apically rounded, with a J­, wedge-shaped, subapical ring, 3±1–3±8 µm (xa ¯ 3±5 µm, n ¯ 10) diam, 1–1±5 µm (xa ¯ 1±4 µm, n ¯ 10) high (Figs 11, 12). Ascospores 12±5–16±5¬5±5–6±5¬4–5 µm (xa ¯ 15±4¬5±9¬4±5 µm, n ¯ 25), overlapping uniseriate, inequilaterally ellipsoidal, with one side flattened, brown, unicellular, smooth-walled, with hyaline, polar mucilaginous pads, 3±1–3±8 µm (xa ¯ 3±5 µm, n ¯ 10) long, 0±5–0±6 µm (xa ¯ 0±6 µm, n ¯ 10) wide, germ slit straight and full length, indistinct (Figs 13–15). Material examined : South Africa : Richards Bay, Meerens, on the stem of Phoenix reclinata, 15 Nov. 1994, K. D. Hyde & T. Steinke (HKU(M) 2109–holotypus). Other material examined : Venezuela : El Limo! n, valle de Puerto La Cruz, D. F., on Olyra latifolia, 20 Jan. 1928, H. Sydow, 311 (S–holotype of A. olyrae)–USA : New York, on Spiraea opulifolia L., ex Herb. Berk., 4428 (K 56361–holotype of A. closterium) ; Desmazie' res, Plantes Cryptogames de France, Edn I, se! r. I, 1825–1851, no. 2075, on Juncus articulatus (NY–isotype of A. tumulosa). Anthostomella applanata is most similar to A. olyrae, which has ascospores surrounded by a thin sheath, rather than mucilaginous pads at the poles, and to A. closterium, which has long polar caudate appendages. It is also distinct from A. tumulosa, which has broadly inequilaterally ellipsoid ascospores, which are larger (17±5–22±5¬6±5–9±5 µm) and without a germ slit. Anthostomella colligata K. D. Hyde & B. S. Lu, sp. nov. (Figs 16–26, 79) Etym. : from the Latin colligata meaning ‘ constriction ’ in reference to the constricted sheath. Ascomata 300–450 µm diam, 340–450 µm alta, subglobosa, ostiolata. Asci 125–200¬9–10 µm, apparatu subapicali cuneato iodo coeru- Key to Anthostomella species in South Africa 1a. Ascospores with a hyaline dwarf cell 1b. Ascospores lacking a dwarf cell . . . . . . . . . . . . . 2a. Ascospores ellipsoidal with one end sharply pointed and the other end truncate, lacking a mucilaginous sheath 2b. Ascospores not sharply pointed at one end or truncate at the other, with a mucilaginous sheath . . . . . . rubicola . . 3 3a. Ascospores 14–22±5¬9–11±5¬6±5–7±5 µm, mucilaginous sheath constricted in the middle 3b. Ascospores 15–17±5¬7±5–9 µm, mucilaginous sheath not constricted . . . . . . . . . . . . . . . colligata .meerensis 4a. Ascospores with echinate wall ornamentations . 4b. Ascospores smooth-walled . . . . . . . . . . . . . . . . . . . . . . . . . . 2 4 . . . . . . . . . . . . . . . . . . . . . . . capensis . . 5 . . . . . . . . . . . . . . . . . . . . . . . . 6a. Ascus without an obvious apical apparatus, ascospores lacking a sheath 6b. Ascus with a subapical ring, ascospores surrounded by a sheath . . . . . . . . . . . . . . . . . . . caffrariae . . 7 7a. Ascal ring non-amyloid (I®) 7b. Ascal ring amyloid (I­) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . raphiae . . 9 . acuminata . applanata 5a. Ascospores with a spiral germ slit . . . . 5b. Ascospores with a straight germ slit or germ slit lacking . . . . . . . . . . . . . . . . 8a. Ascal ring discoid, ascospores 11±5–19¬6±5–7±5 µm, surrounded by a mucilaginous sheath 8b. Ascal ring wedge-shaped, ascospores with hyaline mucilaginous end caps . . . 9a. Asci short-pedicellate, ascospores 12–14±5¬4±5–5¬3±5–4 µm, ellipsoidal with tapering ends ; on Jujubiopsis . 9b. Asci long-pedicellate, ascospores 12±5–16±5¬5±5–6±5¬4–5 µm, inequilaterally ellipsoidal with one-side flattened ; on Phoenix . . . . . . . . . . . . . . . . . . spiralis . 6 palmae . 8 New species of Anthostomella 744 Figs 1–7. Anthostomella acuminata (from holotype). Fig. 1. Ascomata on host surface. Fig. 2. Section of ascoma ; note the blackened clypeus. Fig. 3. Peridium ; note inner hyaline to light coloured, compressed cells and outer darkened, thick-walled cells. Fig. 4. Mature and immature asci. Fig. 5. Apex of ascus ; note J­, wedge-shaped subapical ring. Figs 6, 7. Ascospores, note straight germ slit (arrowed in Fig. 6) and hyaline, mucilaginous end caps (arrowed in Fig. 7). (Bars : Fig. 1 ¯ 1 mm ; Fig. 2 ¯ 50 µm ; Fig. 4 ¯ 10 µm ; Figs 3, 5–7 ¯ 5 µm.) lescente, 4±4–5 µm diam, 1±9–2±5 µm alto praediti. Ascosporae 14–22±5¬9–11±5¬6±5–7±5 µm, late ellipsoideae, unicellulares, tunico mucilaginosa in mediatas constricto praeditae. Ascomata deeply immersed, visible as tiny blackened dots, solitary or clustered (Fig. 16) ; in vertical section 300–450 µm (xa ¯ 350±8 µm, n ¯ 13) diam, 340–450 µm (xa ¯ 393±1 µm, n ¯ 13) high, brown, subglobose, with a central, ostiolar canal and a long ostiole, 35–55 µm (xa ¯ 42±5 µm, n ¯ 8) diam, 90–140 µm (xa ¯ 108±8 µm, n ¯ 8) high (Figs 17, 18). Peridium 15–40 µm (xa ¯ 17 µm, n ¯ 10) wide, comprising several layers of compressed cells, walls light brown at the inside and dark brown and thick-walled towards the outside (Fig. 19). Paraphyses 1±5–3±8 µm (xa ¯ 2±2 µm, n ¯ 10) wide, hyphalike, flexuose and septate. Asci 125–200¬9–10 µm (xa ¯ 161¬9±7 µm, n ¯ 20), (4-)8-spored, cylindrical, longpedicellate, pedicle 20–40 µm (xa ¯ 30±3 µm, n ¯ 20) long, unitunicate, apically rounded, with a J­, wedge-shaped, subapical ring, 4±4–5 µm (xa ¯ 4±9 µm, n ¯ 10) diam, 1±9– 2±5 µm (xa ¯ 2±2 µm, n ¯ 10) high (Figs 20–22). Ascospores 14–22±5¬9–11±5¬6±5–7±5 µm (xa ¯ 17±1¬9±9¬7±3 µm, n ¯ 25), uniseriate, broadly ellipsoidal, brown, unicellular, with a hyaline dwarf cell, 2±5–5 µm (xa ¯ 3±6 µm, n ¯ 12) long, 2±5–3 µm (xa ¯ 2±6 µm, n ¯ 12) wide at the base, smoothwalled, entire spores surrounded by a 2±5–5 µm (xa ¯ 3±7 µm, B.-S. Lu, K. D. Hyde and C. Y. Liew 745 Figs 8–15. Anthostomella applanata (from holotype). Fig. 8. Ascomata on host surface. Fig. 9. Section of ascoma. Fig. 10. Peridium ; note walls hyaline at the inside and dark toward the outside. Fig. 11. Ascus. Fig. 12. Apex of ascus, with J­, wedge-shaped, subapical ring. Figs 13–15. Ascospores, flattened on one side, with hyaline, polar mucilaginous end caps (arrowed in Fig. 13). (Bars : Fig. 8 ¯ 1 mm ; Figs 9, 11 ¯ 10 µm ; Figs 12–15 ¯ 5 µm ; Fig. 10 ¯ 2 µm). n ¯ 15) mucilaginous sheath, which is constricted in the middle, lacking a germ slit (Figs 23–26). Material examined : South Africa : Kwa Zulu-Natal, The National Monument, Mt Unzini, on the stem of Raphia australis, 21 Nov. 1994, K. D. Hyde & T. Steinke (HKU(M) 2160–holotypus). Other material examined : French Guyana, on dead palm petiole, Leprieur (IMI 180622–lectotype of A. sulcigena). Anthostomella colligata is distinguished from other species examined by the constricted mucilaginous sheath of its ascospores. Similar species are A. sulcigena, which has larger spores (20–25¬8–10 µm) with polar mucilage pads but no dwarf cell, and A. formosa which has smaller spores (13±5–15±5¬6–7 µm) and no ascus apical apparatus (Francis 1975). Anthostomella meerensis B. S. Lu & K. D. Hyde, sp. nov. (Figs 27–36, 78) Etym. : from the collection site, Meerens. Ascomata 230–350 µm diam, 320–400 µm alta, globosa, ostiolata. Asci 135–165¬10–15 µm, sine apparatu subapicali iodo, 5–7±5 µm diam, 2–2±5 µm alto praediti. Ascosporae 15–17±5¬7±5–9 µm, late ellipsoideae, cum cella nano hyalino praeditae, fissura germinativa dispansa praeditae. Ascomata deeply immersed, visible as tiny blackened papillae, clustered or mostly solitary (Fig. 27) ; in vertical section 230–350 µm (xa ¯ 302 µm, n ¯ 10) diam, 320–400 µm (xa ¯ 365 µm, n ¯ 10) high, brown, coriaceous, flask-shaped, with a slightly oblique, ostiolar canal and a long ostiole, 15–30 µm (xa ¯ 18±8 µm, n ¯ 8) diam, 90–170 µm (xa ¯ New species of Anthostomella 746 Figs 16–26. Anthostomella colligata (from holotype). Fig. 16. Ascomata on host surface. Note the tiny blackened ostiolar dots. Fig. 17. Section of ascoma. Fig. 18. Ostiolar canal, note the periphyses. Fig. 19. Peridium. Note inner cells which are light coloured and elongate and the outer cells which are darker. Fig. 20. Squash mount of asci and paraphyses. Fig. 21. Asci. Fig. 22. Apex of immature ascus ; note J­, wedge-shaped, subapical ring. Figs 23–26. Ascospores ; note hyaline dwarf cells and sheath which constricts in the middle (arrowed in Fig. 24). (Bars : Fig. 16 ¯ 500 µm ; Figs 17, 20 ¯ 50 µm ; Figs 18, 21 ¯ 20 µm ; Figs 19, 22–26 ¯ 10 µm). 127±5 µm, n ¯ 8) high (Figs 28, 29). Peridium 17±5–30 µm (xa ¯ 21±3 µm, n ¯ 10) wide, comprising several layers of compressed cells, walls hyaline at the inside, and brown and thick-walled towards the outside, fusing externally with the host tissue (Fig. 30). Paraphyses 1±3–3±8 µm (xa ¯ 2±6 µm, n ¯ 10) wide, hypha-like, numerous, hyaline, septate and embedded in a gelatinous matrix (Fig. 31). Asci 135–165¬10–15 µm (xa ¯ 150¬12±3 µm, n ¯ 15), eight- B.-S. Lu, K. D. Hyde and C. Y. Liew 747 Figs 27–36. Anthostomella meerensis (from holotype). Fig. 27. Ascomata on host surface. Fig. 28. Section of ascoma ; note flask-shape and slightly oblique ostiole. Fig. 29. Long periphysate ostiolar canal. Fig. 30. Peridium ; note compressed cells, walls hyaline at the inside and dark and thick-walled towards the outside, fusing externally with host tissue. Fig. 31. Paraphyses embedded in a gelatinous matrix. Fig. 32. Apex of immature ascus ; note J®, wedge-shaped, subapical ring. Fig. 33. Ascus. Figs 34–36. Ascospores ; note dwarf cells (arrowed in Fig. 35) and entire spores surrounded by a mucilaginous sheath. (Bars : Fig. 27 ¯ 500 µm ; Figs 28, 33 ¯ 50 µm ; Fig. 31 ¯ 20 µm ; Figs 29, 30, 32, 34–36 ¯ 10 µm). spored, cylindrical, short-pedicellate, unitunicate, apically rounded, with a J®, wedge-shaped, subapical ring, 5–7±5 µm (xa ¯ 6±3 µm, n ¯ 10) diam, 2–2±5 µm (xa ¯ 2±4 µm, n ¯ 10) high (Figs 32, 33). Ascospores 15–17±5¬7±5–9 µm (xa ¯ 15±9¬7±9 µm, n ¯ 25), uniseriate, broadly ellipsoidal, dark brown or brown, unicellular, with a hyaline dwarf cell, 1±9–3±1 µm (xa ¯ 2±5 µm, n ¯ 10) long, 2±5–3±1 µm (xa ¯ 2±8 µm, n ¯ 10) wide at the base, smooth-walled, entire spores New species of Anthostomella 748 Figs 37–43. Anthostomella palmae (from holotype). Fig. 37. Ascomata on host surface. Fig. 38. Section of ascoma. Fig. 39. Peridium comprising compressed cells, walls hyaline at the inside and dark and thick-walled towards the outside. Fig. 40. Asci, with J®, discoid, subapical ring. Figs 41–43. Ascospores ; note hyaline mucilaginous sheath (in India Ink in Fig. 41). (Bars : Fig. 37 ¯ 1 mm ; Fig. 38 ¯ 50 µm ; Figs 40–43 ¯ 10 µm ; Fig. 39 ¯ 5 µm). surrounded by a 1–1±5 µm (xa ¯ 1±3 µm, n ¯ 10) mucilaginous sheath, germ slit straight, extending over the full length, indistinct (Figs 34–36). Anthostomella palmae K. D. Hyde & B. S. Lu, sp. nov. (Figs 37–43, 76) Etym. : in reference to the palm host. Material examined : South Africa : Richards Bay, Meerens, on the stem of Phoenix reclinata, 15 Nov. 1994, K. D. Hyde & T. Steinke (HKU(M) 2134–holotypus). Ascomata 150–260 µm diam, 200–260 µm alta, globosa, ostiolata. Asci 85–113¬11±3–15 µm, (6–)eight-spori, sine apparatu subapicali iodo, 3±1–8±8 µm diam, 0±6–1±3 µm alto praediti. Ascosporae 15–19¬7±5–10¬5–6±5 µm, late ellipsoideae, unilateraliter applanatae. Other material examined : USA : South Carolina, Car. Inf., on Smilax laurifolia, 1855, no. 4877, ex Herb. Berk. (K 56336–holotype of A. sepelibilis). Anthostomella meerensis is similar to A. formosa, which has slightly smaller ascospores (13±5–15±5¬6–7 µm) and no ascus apical apparatus (Francis 1975), and A. sepelibilis, which is clypeate, has a J­subapical ascus ring, and ascospores without a germ slit. Ascomata immersed, visible as darkened areas, clustered or mostly solitary (Fig. 37) ; in vertical section 150–260 µm (xa ¯ 208 µm, n ¯ 10) diam, 200–260 µm (xa ¯ 230 µm, n ¯ 10) high, brown, coriaceous, globose, with a central, periphysate ostiolar canal, 25–40 µm (xa ¯ 32±5 µm, n ¯ 10) diam, 40– 60 µm (xa ¯ 51 µm, n ¯ 10) high (Fig. 38). Clypeus black, ca 250 µm diam, 20 µm high, comprising host cells mixed with blackened intracellular fungal hyphae (Fig. 38). Peridium B.-S. Lu, K. D. Hyde and C. Y. Liew 749 Figs 44–52. Anthostomella raphiae (from holotype). Fig. 44. Ascomata on host surface. Fig. 45. Section of ascoma. Fig. 46. Periphysate ostiole. Fig. 47. Peridium of compressed cells, walls hyaline at the inside, and dark and thick-walled towards the outside. Fig. 48. Ascus. Fig. 49. Apex of ascus ; note the weakly J­, discoid, subapical ring. Figs 50–52. Ascospores. Note mucilaginous sheath, which is thicker in the middle. (Bars : Fig. 44 ¯ 200 µm ; Fig. 45 ¯ 50 µm ; Figs 46, 48 ¯ 20 µm ; Figs 49–52 ¯ 10 µm ; Fig. 47 ¯ 5 µm). 11±3–17±5 µm (xa ¯ 14±6 µm, n ¯ 10) wide, comprising several layers of compressed cells, walls hyaline at the inside and brown and thick-walled towards the outside (Fig. 39). Paraphyses 1±3–3±1 µm (xa ¯ 2±5 µm, n ¯ 10) wide, hyphalike, flexuose, septate, numerous and embedded in a gelatinous matrix. Asci 85–113¬11±3–15 µm (xa ¯ 104±5¬12±7 µm, n ¯ 15), (6-)eight-spored, cylindrical, short-pedicellate, unitunicate, apically rounded, with a J®, discoid, subapical ring, 3±1–8±8 µm (xa ¯ 4 µm, n ¯ 10) diam, 0±6–1±3 µm (xa ¯ 0±9 µm, n ¯ 10) high (Fig. 40). Ascospores 15–19¬7±5–10¬5–6±5 µm (xa ¯ 17±9¬8±8¬5±8 µm, n ¯ 25), overlapping uniseriate, broadly ellipsoidal, with one side flattened, dark brown, unicellular, surrounded by a 1–1±5 µm (xa ¯ 1±3 µm, n ¯ 10) thick mucilaginous sheath, germ slit straight, extending over the full length, ventral (Figs 41–43). Material examined : South Africa : Richards Bay, Meerens, on the stem of Phoenix reclinata, 21 Nov. 1994, K. D. Hyde & T. Steinke (HKU(M) 2209–holotypus). Other material examined : Desmazie' res, Plantes Cryptogames de France, Edn I, se! r. I, 1825–1851, no. 2075, on Juncus articulatus (NY– isotype of A. tumulosa.) New species of Anthostomella 750 Figs 53–61. Anthostomella caffrariae (from holotype). Fig. 53. Ascomata on host surface. Fig. 54. Section of ascoma. Fig. 55. Periphysate ostiolar canal. Fig. 56. Peridium, comprising dark, thick-walled, elongate cells. Fig. 57. Ascus. Fig. 58. Apex of ascus ; note lack of apical apparatus. Figs 59–61. Ascospores, in face view (in Fig. 59) and side view (in Fig. 60). (Bars : Fig. 53 ¯ 200 µm ; Figs 54–56 ¯ 20 µm ; Figs 57–61 ¯ 10 µm). Anthostomella palmae is distinct from A. dilatata because ascospores in A. palmae have more tapering ends, while those of A. dilatata have rounded ends, and are shorter and narrower compared to the ascospores of A. dilatata (15–19¬7±5–10¬ 5–6±5 µm in A. palmae, while 16–22¬10–15¬5–6 µm in A. dilatata) (Hyde 1996 : 296). Anthostomella palmae differs from A. tumulosa because ascospores in A. palmae have rounded ends, and are smaller (15–19¬7.5–10¬5–6.5 µm, xa ¯ 17.9¬8.8¬5.8 µm), while those of A. tumulosa have tapering ends, and are relatively larger (17.5–22.5¬6.5– B.-S. Lu, K. D. Hyde and C. Y. Liew 9.5 µm, xa ¯ 20.3¬7 µm). Anthostomella palmae is distinguished by its smaller ascospores from A. dilatata (16–22¬10– 15¬5–6 µm) with rounded ends (Hyde 1996), and A. tumulosa (17±5–22±5¬6±5–9±5 µm). The latter, also has an I­ wedgeshaped subapical ascal ring. Anthostomella raphiae B. S. Lu & K. D. Hyde, sp. nov. (Figs 44–52, 75) Etym. : in reference to the host. Ascomata 270–370 µm diam, 250–350 µm alta, subglobosa, ostiolata. Asci 90–143¬12±5–15 µm, apparatu subapicali iodo coerulescente, 4±4–6±3 µm diam, 0±6–1±9 µm alto praediti. Ascosporae 11±5–19¬6±5–7±5 µm, inaequilateraliter ellipsoideae, tunica mucilaginosa in medietas incrassato et fissura germinativa dispansa praeditae. Ascomata immersed, visible as blackened, raised, domeshaped areas, 0±21–0±36¬0±28–0±42 mm (xa ¯ 0±31¬ 0±36 mm, n ¯ 10), clustered or mostly solitary (Fig. 44) ; in vertical section 270–370 µm (xa ¯ 336±2 µm, n ¯ 15) diam, 250–350 µm (xa ¯ 300 µm, n ¯ 15) high, brown, coriaceous, subglobose, with a central periphysate canal, 20–40 µm (xa ¯ 33±3 µm, n ¯ 10), 80–110 µm (xa ¯ 87±5 µm, n ¯ 10) high (Figs 45, 46). Clypeus black, ca 400 µm diam, 50 µm high near the neck, comprising host cells mixed with blackened intracellular fungal hyphae (Fig. 45). Peridium 8±8–18±8 µm (xa ¯ 14±4 µm, n ¯ 10) wide, comprising several layers of compressed cells, walls hyaline at the inside, and brown and thick-walled towards the outside (Fig. 47). Paraphyses 1±3–3±8 µm (xa ¯ 2±8 µm, n ¯ 15) wide, hypha-like, flexuose, numerous, septate and embedded in a gelatinous matrix. Asci 90–143¬12±5–15 µm (xa ¯ 121±6¬13±7 µm, n ¯ 20), eightspored, cylindrical, short-pedicellate, unitunicate, apically rounded, with a weakly J­, discoid, subapical ring, 4±4–6±3 µm (xa ¯ 5 µm, n ¯ 10) diam, 0±6–1±9 µm (xa ¯ 1±1 µm, n ¯ 10) high (Figs 48, 49). Ascospores 11±5–19¬6±5–7±5 µm (xa ¯ 15±4¬6±9 µm, n ¯ 25), partly overlapping uniseriate, inequilaterally ellipsoidal, dark brown, unicellular, smoothwalled, surrounded by a 2±5–3±8 µm (xa ¯ 2±8 µm, n ¯ 25) mucilaginous sheath, which is thinner at the poles, 1–2±5 µm (xa ¯ 1±4 µm, n ¯ 10) thick, germ slit full length, straight (Figs 50–52). Material examined : South Africa : Kwa Zulu-Natal, The National Monument, Mt. Unzini, on the bark of Raphia australis, 21 Nov. 1994, K. D. Hyde & T. Steinke (HKU(M) 2146–holotypus). Other material examined : Austria : Tirolia, Kastelruth, on Labiatae sp., 8 Sep. 1903, H. Rehm (S–holotype of Anthostomella megaclypeata)– Bavaria : Kampenwand, on stem of Compositae sp., Sep. 1904, H. Rehm (S–holotype of Anthostomella subconica). Anthostomella raphiae is distinct from A. megaclypeata and A. subconica, which have wedge-shaped subapical rings, narrower, long-pedicellate asci (9–11±5 µm) and lighter brown ascospores with only a very thin sheath. It is also distinguished by its ascospore shape from A. conorum, which has broadly ellipsoidal spores 13–17¬7–9 µm (Francis 1975). Anthostomella caffrariae B. S. Lu & K. D. Hyde, sp. nov. (Figs 53–61, 77) Etym. : from the Latin caffrariae meaning ‘ South Africa ’, in reference to the collection site of South Africa. 751 Ascomata 190–310 µm diam, 180–230 µm alta, subglobosa, ostiolata. Asci 82–110¬11±3–15 µm, apparatu apicali non praediti. Ascosporae 14–17±5¬9–11±5¬5–75 µm, late inaequilateraliter ellipsoideae, fissura germinativa dispansa praeditae. Ascomata immersed, visible as tiny blackened dots, solitary (Fig. 53) ; in vertical section 190–310 µm (xa ¯ 248 µm, n ¯ 10) diam, 180–230 µm (xa ¯ 209 µm, n ¯ 10) high, brown, coriaceous, subglobose, with a central periphysate ostiolar canal, 20–30 µm (xa ¯ 28 µm, n ¯ 6) diam, 40–80 µm (xa ¯ 64 µm, n ¯ 6) high (Figs 54, 55). Peridium 5–13±8 µm (xa ¯ 9±3 µm, n ¯ 10) wide, comprising several layers of brown, thick-walled, elongate cells (Fig. 56). Paraphyses 1–1±9 µm (xa ¯ 1±5 µm, n ¯ 10) wide, hypha-like, flexuose, numerous and septate. Asci 82–110¬11±3–15 µm (xa ¯ 98±9¬13±7 µm, n ¯ 20), eight-spored, broad cylindrical, short-pedicellate, unitunicate, apically rounded, lacking an apical apparatus (Figs 57, 58). Ascospores 14–17±5¬9–11±5¬5–7±5 µm (xa ¯ 16±8¬ 9±8¬5±8 µm, n ¯ 25), overlapping uniseriate, broadly inequilaterally ellipsoidal in face view, oblong-ellipsoidal in side view, dark brown, unicellular, smooth-walled, germ slit straight, extending over the full length (Figs 59–61). Material examined : South Africa : Richards Bay, Meerens, on the stem of Phoenix reclinata, 15 Nov. 1994, K. D. Hyde & T. Steinke (HKU(M) 2140–holotypus). Other material examined : Philippines : Los Ban4 os, on stems of Corypha elata, 30 Apr. 1913, Evaristo 2572 (S–holotype of A. coryphae f. minutussima). Anthostomella caffrariae is distinguished from A. coryphae f. minutissima, which has slightly wider ascospores (6±6–9 µm) with a sheath, from A. pedemontana which has globose ascomata, slightly smaller ascospores (13–16¬6–7 µm), and a germ slit which does not run the full length of the spore (Francis 1975), and from A. dilatata, which has broadly ellipsoid, almost rounded, and larger ascospores (16–22¬10–15¬5–6 µm) (Hyde 1996). Anthostomella spiralis K. D. Hyde & B. S. Lu, sp. nov. (Figs 62–71, 73) Etym. : from the Latin spiralis meaning ‘ spiral ’ in reference to the spiral germ slit. Ascomata 170–285 µm diam, 165–275 µm alta, subglobosa, ostiolata. Asci 95–143¬9±5–11±3 µm, apparatu subapicali iodo coerulescente, 3±1–5 µm diam, 0±5–1±5 µm alto. Ascosporae 12– 14±5¬5–5±5 µm, late inaequilateraliter ellipsoidae, tunica mucilaginosa in medietas incrassato et fissura germinativa helicoideo praeditae. Ascomata immersed, visible as blackened areas, solitary or clustered (Fig. 62) ; in vertical section 170–285 µm (xa ¯ 234±4 µm, n ¯ 10) diam, 165–275 µm (xa ¯ 221±1 µm, n ¯ 10) high, dark brown, coriaceous, subglobose, with a central, periphysate ostiolar canal, 12±5–20 µm (xa ¯ 16 µm, n ¯ 10) diam, 15–30 µm (xa ¯ 22±5 µm, n ¯ 10) high (Figs 63, 64). Peridium 7±5–14±5 µm (xa ¯ 10±1 µm, n ¯ 10) wide, comprising several layers of dark brown-walled elongate cells, thin at the base (Fig. 65). Paraphyses 2±5–3±8 µm (xa ¯ 3±1 µm, n ¯ New species of Anthostomella 752 Figs 62–71. Anthostomella spiralis (from holotype). Fig. 62. Ascomata on host surface. Fig. 63. Section of ascoma ; note peridium which is thin at the base. Fig. 64. Ostiole, with periphyses. Fig. 65. Peridium, comprising several layers of dark-walled, elongate cells. Fig. 66. Asci. Fig. 67. Apex of ascus ; note J­, discoid, subapical ring. Fig. 68. Squash mount of asci and paraphyses. Figs 69–71. Ascospores, note spiral germ slits (arrowed in Fig. 69), surrounded by thick mucilaginous sheath, which is thicker in the centre. (Bars : Fig. 62 ¯ 500 µm ; Figs 63, 68 ¯ 50 µm ; Fig. 64 ¯ 20 µm ; Figs 66, 69–71 ¯ 10 µm ; Figs 65, 67 ¯ 5 µm). B.-S. Lu, K. D. Hyde and C. Y. Liew 72 73 74 75 76 77 78 79 753 A comparison of A. spiralis with other species of Anthostomella whose ascospores have a spiral germ slit is provided in Table 1. Hawksworth & Lodha (1983) introduced Helicogermslita for Anthostomella-like species whose ascospores have a spiral germ slit. Currently there are five species in this genus (Hawksworth et al. 1995), but it has been much debated (Rappaz 1995). Dargan et al. (1984) suggested a possible relationship between Helicogermslita and Rosellinia because the type species, H. celastri, has ascospores with a dwarf cell and asci with a subapical ring which is sometimes amyloid. Petrini (1993), however, did not treat Helicogermslita as a synonym of Rosellinia. Læssøe et al. (1994) included four species in Helicogermslita and Rappaz (1995) accepted the genus and described H. fleischhackii. We treat Anthostomella spiralis in Anthostomella instead of in Helicogermslita because A. spiralis lacks stromata, unlike the species in Helicogermslita. Figs 72–79. Ascospores of Anthostomella. Fig. 72. A. acuminata. Fig. 73. A. spiralis. Fig. 74. A. applanata. Fig. 75. A. raphiae. Fig. 76. A. palmae. Fig. 77. A. caffrariae. Fig. 78. A. meerensis. Fig. 79. A. colligata. Bar ¯ 10 µm. 10) wide, hypha-like, flexuose, numerous and septate (Fig. 68). Asci 95–142±5¬9±5–11±3 µm (xa ¯ 107±4¬10±5 µm, n ¯ 20), eight-spored, cylindrical, short-pedicellate, unitunicate, apically rounded, with a J­, discoid, subapical ring, 3±1–5 µm (xa ¯ 4 µm, n ¯ 10) diam, 0±5–1±5 µm (xa ¯ 0±8 µm, n ¯ 10) high (Figs 66–68). Ascospores 12–14±5¬5–5±5 µm (xa ¯ 13±2¬5±1 µm, n ¯ 25), overlapping uniseriate, broadly inequilaterally ellipsoidal, dark brown, unicellular, smoothwalled, surrounded by a 1±9–2±8 µm (xa ¯ 2±1 µm, n ¯ 10) mucilaginous sheath, which is thicker in the middle, germ slit spiral (Figs 69–71). Material examined : South Africa : Richards Bay, Meerens, on the stem of Phoenix reclinata, 15 Nov. 1994, K. D. Hyde & T. Steinke (HKU(M) 2119–holotypus ; HKU(M) 2109 ; HKU(M) 2106). A C K N O W L E D G E M E N TS Bing-Sheng Lu would like to thank The University of Hong Kong for the award of a Postgraduate Studentship. Dr T. K. Goh is thanked for his help in naming species and correcting the Latin diagnosis. Dr K. D. Hyde is grateful to Dr T. S. Steinke for funding, arranging and accompanying him on the visit to South Africa. REFERENCES Dargan, J. S., Singh, M. & Rogers, J. D. (1984) A note on Helicogermslita celastri. Mycologia 76 : 1113–1115. Doidge, E. M. (1948) South African ascomycetes in the National Herbarium. Bothalia 4 : 837–893. Francis, S. M. (1975) Anthostomella Sacc. (Part I). Mycological Papers 139 : 1–97. Hawksworth, D. L., Kirk, P. M., Sutton, B. C. & Pegler, D. N. (1995) Ainsworth & Bisby’s Dictionary of the Fungi. 8th Edn, CAB International : Cambridge. Hawksworth, D. L. & Lodha, B. C. (1983) Helicogermslita, a new stromatic xylariaceous genus with a spiral germ slit from India. Transactions of the British Mycological Society 81 : 91–96. Table 1. Comparison of Anthostomella species whose ascospores have spiral germ slit. Hosts Clypeus Ascomata Asci Ascospores A. adusta Salix No Globose to subglobose, 420–630¬480–590 µm A. cassinopsidis Cassinopsis Yes Globose, 110–160¬ 140–170 µm A. limitata Yes Globose, 110–160¬ 140–170 µm A. semele Umbelliferae, Cornus, Daemonorops, Callistemon Semele 125–185¬12±5–15 µm, wedge- Ellipsoidal, 17±5–25¬6±5–8 µm, shaped J­ subapical ring, with a dwarf cell 2±5–3±1¬2±5–3±1 µm 112±5–130¬13±8–15 µm, Broadly ellipsoidal, 20–34¬ discoid J­ subapical ring, 11±5–14¬8–10±5 µm, 3±8–5¬0±6–1±5 µm thin sheath 70–83±8¬6±9–8±1 µm, discoid Ellipsoidal, 10–14±5¬4±5–7±5 µm, J­ subapical ring, thin sheath 2±3–2±5¬0±6–1±3 µm No Globose, 200¬210 µm Not seen A. spiralis Phoenix No Subglobose, 170–285¬ 165–275 µm 95–142±5¬9±5–11±3 µm, discoid J­ subapical ring, 3±1–5¬0±5–1±5 µm Oblong-ellipsoidal, with one side flattened, 10–15¬4±5 µm Broadly inequilaterally ellipsoidal with tapering end, 12–14±5¬5–5±5 µm, thick sheath New species of Anthostomella Hyde, K. D. (1996) Fungi from palms. XXVI. The genus Anthostomella, with ten new species. Nova Hedwigia 62 : 273–340. Læssøe, T. & Spooner, B. M. (1994) Rosellinia and Astrocystis (Xylariaceae) : new species and genetic concepts. Kew Bulletin 49 : 1–70. Martin, P. (1969) Studies in the Xylariaceae : VII. Anthostomella and Lopadostoma. The Journal of South African Botany 35 : 393–410. 754 Petrini, L. E. (1993) Rosellinia species of the temperate zone. Sydowia 44 : 169–281. Rappaz, F. (1995) Anthostomella and related xylariaceous fungi on hard wood from Europe and North America. Mycologia Helvetica 7 : 99–168. Corresponding Editor : C. A. Shearer