Colletotrichum: complex species or species ... - CBS - KNAW

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Damm et al. Fig. 7. Colletotrichum cosmi (from ex-holotype strain CBS 853.73). A–B. Conidiomata. C–H. Conidiophores. I–N. Appressoria. O–P. Conidia. A, C–E, O. from Anthriscus stem. B, F–N, P. from SNA. A–B. DM, C–P. DIC, Scale bars: A = 200 µm, B = 100 µm, C = 10 µm. Scale bar of C applies to C–P. Culture characteristics: Colonies on SNA flat with entire margin, hyaline, buff to pale honey, on filter paper partly pale olivaceous grey, the medium, filter paper and Anthriscus stem partly covered with floccose-felty whitish to pale olivaceous grey aerial mycelium and orange acervuli, reverse of filter paper partly pale cinnamon, pale olivaceous grey to olivaceous grey; growth rate 23–24 mm in 7 d (33.5–34 mm in 10 d). Colonies on OA flat with entire margin; surface entirely covered with thin floccose-felty white to pale olivaceous grey aerial mycelium and orange acervuli, reverse vinaceous buff, purplish grey to fuscous black; growth rate 23–25 mm in 7 d (33.5–37.5 mm in 10 d). Conidia in mass orange. Material examined: Netherlands, Wageningen, from seed of Cosmos sp., collection date and collector unknown (deposited in CBS collection in Nov. 1973 by G.H. Boerema), (CBS H-20794 holotype, culture ex-type CBS 853.73 = PD 73/856). Notes: Kwon et al. (1999) report C. acutatum (s. lat.) to cause sunken brownish spots on stems, as well as symptoms on leaves, flowers and floral axes of Cosmos bipinnatus in Korea. Morphological characters (conidia, appressoria) are similar to those of strain CBS 853.73, except for setae, which our strain did not develop in our standard culture conditions. It is therefore possible that the collection from Korea represents C. cosmi. Colletotrichum acutatum (s. lat.) is also known as an anthracnose pathogen of flowers and flower buds of Cosmos bipinnatus in Japan (Yaguchi et al. 1996). The shape of conidia and appressoria of the Japanese fungus are similar to our strain, but the conidia are smaller, measuring 11–14 × 2.8–3.5 µm. Two other species are reported from Cosmos bipinnatus in India, C. truncatum (as C. capsici) associated with seeds and causing seed and seedling rot (Srivastava et al. 1981) and C. gloeosporioides associated with leaves (Kumari et al. 1981). When strain CBS 853.73 was first accessed into CBS, von Arx identified it as C. gloeosporioides, but with the remark “deviating by longer, slender conidia”. Molecular data do not support this identification; the strain belongs to the C. acutatum complex, but it is possible that reports of C. gloeosporioides refer to this species. Colletotrichum cosmi is part of clade 2. It can be separated from other species by all gene sequences, but mostly with only 1 bp divergence. There are more sequence divergences in GAPDH and HIS3; however, with these genes individually, the species sits within the very variable C. nymphaeae clade. The closest match in a blastn search with the TUB2 sequence of strain CBS 853.73 (with 99 % identity, 4 bp differences) was GU246633 from isolate R14 from Capsicum annuum from South Korea (Sang et al. 2011), while the closest match with the GAPDH sequence covering ± the full length sequence (with 98 % identity, 4 bp differences) was HQ846724 from isolate OBP6 from an unknown plant, probably from India (P. Chowdappa, C.S. Chethana, S. Madhura, unpubl. data). We do not consider that these data in isolation are sufficient evidence to identify these sequences as originating from C. cosmi. There are 22 sequences in GenBank that match the ITS sequence of strain CBS 853.73 with 99 % identity, all with 2 bp differences. 62
The Colletotrichum acutatum species complex Fig. 8. Colletotrichum costaricense (from ex-holotype strain CBS 330.75). A–B. Conidiomata. C–H. Conidiophores. I–N. Appressoria. O–P. Conidia. A, C–D, O. from Anthriscus stem. B, E–N, P. from SNA. A–B. DM, C–P. DIC, Scale bars: A = 100 µm, C = 10 µm. Scale bar of A applies to A–B. Scale bar of C applies to C–P. Colletotrichum costaricense Damm, P.F. Cannon & Crous, sp. nov. MycoBank MB800499. Fig. 8. Etymology: Named after the country where it was collected, Costa Rica. Sexual morph not observed. Asexual morph on SNA. Vegetative hyphae 1–9.5 µm diam, hyaline, smooth-walled, septate, branched. Chlamydospores not observed. Conidiomata not developed, conidiophores formed directly on hyphae. Setae not observed. Conidiophores hyaline, smooth-walled, simple or septate and branched. Conidiogenous cells hyaline, smooth-walled, cylindrical to ampulliform, polyphialides observed, 4.5–24 × 2–3.5 µm, opening 0.5–1.5 µm diam, collarette 0.5–1.5 µm long, periclinal thickening visible. Conidia hyaline, smooth-walled, aseptate, straight, cylindrical with both ends acute, (9–)11.5–18(–28) × (3–)3.5–4(–4.5) µm, mean ± SD = 14.6 ± 3.1 × 3.7 ± 0.3 µm, L/W ratio = 4.0. Appressoria sparse, single or in small groups, pale brown, smooth-walled, subglobose to elliptical, the edge entire to undulate, (4.5–)6–8.5(–10) × (3–)4–6(–6.5) µm, mean ± SD = 7.1 ± 1.2 × 4.9 ± 0.9 µm, L/W ratio = 1.4, appressoria of strain CBS 211.78 are medium brown. Asexual morph on Anthriscus stem. Conidiomata not developed, conidiophores and setae formed directly on hyphae. Setae medium to dark brown, smooth-walled to finely verruculose, 0–2-septate, 50–60 µm long, base cylindrical, 3.5–4.5 µm diam, the tip ± acute. Conidiophores hyaline to pale brown, smooth-walled, septate, branched, to 40 µm long. Conidiogenous cells hyaline to pale brown, smooth-walled, cylindrical to ampulliform, 8–22 × 3–5 µm, opening 1–1.5 µm diam, collarette 1–1.5 µm long, periclinal thickening distinct. Conidia hyaline, smooth-walled, aseptate, straight, cylindrical with both ends acute, (12.5–)13.5–16(–18) × 3.5–4 µm, mean ± SD = 14.8 ± 1.4 × 3.8 ± 0.3 µm, L/W ratio = 3.9. Culture characteristics: Colonies on SNA flat with entire margin, hyaline to pale cinnamon, on Anthriscus stem partly olivaceous grey to iron-grey, on filter paper pale olivaceous grey to olivaceous grey, with short or woolly white aerial mycelium and few salmon acervuli on filter paper and on Anthriscus stem, reverse of filter paper same colours; growth rate 19–22.5 mm in 7 d (31–34 mm in 10 d). Colonies on OA flat with entire margin; surface olivaceous with pale olivaceous grey to olivaceous grey sectors, the sectors covered with short white aerial mycelium and salmon acervuli or culture completely covered with short felty whitish aerial mycelium, reverse honey, olivaceous grey to iron-grey, growth rate 22–23 mm in 7 d (28.5–34.5 mm in 10 d). Conidia in mass salmon to saffron. Material examined: Costa Rica, Meseta Central, from berry of Coffea arabica cv. Typica, collection date and collector unknown (deposited in CBS collection Jun. 1975 by D. Mulder, Wageningen), (CBS H-20811 holotype, culture ex-type CBS 330.75); Turrialba, from twig of Coffea sp., collection date and collector unknown (deposited in CBS collection Apr. 1978 by C. Bianchini), culture CBS 211.78 = IMI 309622. www.studiesinmycology.org 63
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