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

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Damm et al.Fig. 25. Colletotrichum pyricola (from ex-holotype strain CBS 128531). A–B. Conidiomata. C–K. Conidiophores. L–Q. Appressoria. R–S. Conidia. A, C–F, R. from Anthriscusstem. B, G–Q, S. from SNA. A–B. DM, C–S. DIC, Scale bars: A = 200 µm, C = 10 µm. Scale bar of A applies to A–B. Scale bar of C applies to C–S.restricted terminal growth and increased growth of lateral shoots.Two strains with differing colony characteristics were isolated andwere used in pathogenicity tests on 4-month-old seedlings of Pinusradiata; 11.5 % of the seedlings inoculated with the salmon orangeculture and 92 % of the seedlings with the grey culture showedthe symptoms. The two strains were sent to CBS, and both wereidentified as C. acutatum f. pineum by von Arx, presumably becauseof the identity of the host plant and some of their morphologicalfeatures, especially the conidia with acute ends typical for C.acutatum. One of the strains was kept in the CBS collection asCBS 436.77; unfortunately we can only suppose it was the salmonorange culture.Strain CBS 436.77 turns out not to be closely related to C.acutatum f. pineum, which belongs to C. acutatum s. str. (Fig. 1).Colletotrichum pseudoacutatum is at best basal to the C. acutatumspecies complex and forms a sister group to a clade containing theC. acutatum complex and C. orchidophilum (fig. 2 in Cannon et al.2012, this issue). The closest matches in a blastn search on theITS sequence of strain CBS 436.77 (with only 94 % identity) areunidentified Colletotrichum isolates, e.g. from Podocarpaceae inNew Zealand (Joshee et al. 2009), plus several C. coccodes strainsincluding the ex-epitype strain CBS 164.49 (HM171678, Liu et al.2011), C. trichellum strains MEP1535 (= CBS 118198, DQ286152,Farr et al. 2006) and DAOM 188792 (= CBS 125343, EU400142,wrongly identified as C. dematium, Chen YY, Conner R, BabcockC, Penner W, unpubl. data) and “C. gloeosporioides” strains DAOM183087 (EU400145, probably C. coccodes, Chen YY, Conner R,Babcock C, Penner W, unpubl. data) and BBA 71369 from Pleione(AJ301980, probably C. orchidophilum, Nirenberg et al. 2002).The closest matches with the TUB2 sequence showed only 82 %identity, including C. trichellum strains HKUCC 10378, CBS 217.64and CBS 118198 (GQ849447, Yang et al. 2009, GU228106,GU228107, Damm et al. 2009). There is no match over the wholespan of the GAPDH sequence of this species.In morphological terms, C. pseudoacutatum mainly differs fromspecies in the C. acutatum complex by the formation of pale brown,verruculose, irregular shaped appressoria, and also by the moreabundant formation of setae.Colletotrichum pyricola Damm, P.F. Cannon & Crous, sp.nov. MycoBank MB800510. Fig. 25.Etymology: Named after the host plant Pyrus communis.Sexual morph not observed. Asexual morph on SNA. Vegetativehyphae 1–8 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, septate, branched.Conidiogenous cells hyaline smooth-walled, cylindrical, 9–25 ×2.5–3.5 µm, opening 1–1.5 µm diam, collarette 1–1.5 µm long,periclinal thickening visible. Conidia hyaline, smooth-walled,94
The Colletotrichum acutatum species complexaseptate, straight, fusiform to cylindrical with one end slightly acuteand one end round or slightly acute, (10–)14.5–18.5(–24) × (3.5–)4.5–5(–5.5) µm, mean ± SD = 16.7 ± 2.1 × 4.7 ± 0.4 µm, L/W ratio= 3.5. Appressoria single or in small dense clusters, pale brown,smooth-walled, ellipsoidal, clavate to cylindrical, the edge entire orundulate, (4.5–)6–16(–22) × (3.5–)4.5–7(–8.5) µm, mean ± SD =11.1 ± 5.1 × 5.7 ± 1.2 µm, L/W ratio = 2.0.Asexual morph on Anthriscus stem. Conidiomata acervular,conidiophores formed on pale brown, angular, basal cells 3–7µm diam. Setae not observed. Conidiophores hyaline, smoothwalled,septate, branched, to 40 µm long. Conidiogenous cellshyaline, smooth-walled, cylindrical, 11–20 × 3–4 µm, opening1.5–2 µm diam, collarette 0.5–2 µm long, periclinal thickeningdistinct. Conidia hyaline, smooth-walled, aseptate, straight,fusiform to cylindrical with both ends acute, (9.5–)14–17(–18.5)× (4–)4.5–5(–5.5) µm, mean ± SD = 15.4 ± 1.6 × 4.8 ± 0.4 µm,L/W ratio = 3.2.Culture characteristics: Colonies on SNA flat with entire margin,hyaline, on medium, filter paper and Anthriscus stem partly coveredwith thin floccose white to pale grey aerial mycelium and orangeacervuli, reverse hyaline with orange to grey acervuli shiningthrough; growth rate 24–25 mm in 7 d (35–37 mm in 10 d). Colonieson OA flat to raised with entire margin; surface partly covered withfloccose to woolly whitish to pale olivaceous grey aerial myceliumand orange acervuli mainly appearing in growth rings, reverse buff,olivaceous buff to grey olivaceous with olivaceous grey to irongrey rings; growth rate 21.5–22.5 mm in 7 d (35–37.5 mm in 10 d).Conidia in mass orange.Material examined: New Zealand, WO, Waikato, from fruit rot of Pyrus communis,1 Jun. 1988, unknown collector (deposited in ICMP collection by P.R. Johnston),(CBS H-20810 holotype, culture ex-type CBS 128531 = ICMP 12924 = PRJ 977.1).Notes: This is a third species within clade 4, not clearly distinctfrom C. johnstonii using morphological or cultural characteristicsbut with unique ACT, TUB2, CHS-1, GAPDH and HIS3 sequences.The ITS sequence of C. pyricola is identical with those of C. salicis,C. johnstonii and C. phormii.As with C. johnstonii, C. pyricola appears to be endemic to NewZealand, but more data are needed to confirm its distribution. StrainCBS 128531 (= PRJ 977.1) is the only strain of this species availableto us and was included in C. acutatum group C by Johnston & Jones(1997) and Lardner et al. (1999) and in group F2 by Guerber et al.(2003). In the combined GS and GAPDH phylogeny in Guerberet al. (2003), there is a second strain grouping with C. pyricolathat they assigned as the only representative of their F5 group.This strain (PRJ 823) however belongs to group B in Lardner et al.(1999), with a completely different RAPD banding pattern.In contrast to apple, for which Colletotrichum species are listedas major pathogens causing bitter rot (González et al. 2006), peartrees seem to be rarely affected by anthracnose. Colletotrichum piriNoack was actually described from apple (listed as Pyrus malus,a synonym of Malus pumila) in Brazil, rather than from pear as itsname suggests.The closest match in a blastn search with the TUB2 sequenceof strain CBS 128531 (with 98 % identity, 10 bp differences) wereAJ409294 isolate 90 from Fragaria in the UK (Talhinhas et al. 2002)as well as AJ748609, AJ748612–AJ748614, AJ748619–AJ748622,AJ748625 from olive isolates (Talhinhas et al. 2005). With theGAPDH sequence there was no closer match than 89 % identity.Colletotrichum rhombiforme Damm, P.F. Cannon & Crous,sp. nov. MycoBank MB800511. Fig. 26.Etymology: Named after the shape of the ascospores, which canbe rhomboidal.Sexual morph developed on Anthriscus stem. Ascomata globoseto subglobose, pale brown, 300–400 × 400–500 µm, glabrous,ostiolate. Peridium 8–14 µm thick, composed of pale to mediumbrown flattened angular cells, 6–16 µm diam. Ascogenous hyphaehyaline, smooth-walled, delicate. Interascal tissue composed ofparaphyses, hyaline, septate, branched at the base, 35–80 × 3–5µm, widest part at the base, tips round. Asci cylindrical, 55–73× 9–11 µm, 8-spored. Ascospores arranged uni-to bi-seriately,aseptate, hyaline, smooth-walled, oval, fusiform, or rhomboidal,one end ± acute and one end round or both ends round, sometimesslightly curved, (11–)12.5–16(–17) × 4–)4.5–6(–7.5) µm, mean ±SD = 14.1 ± 1.6 × 5.2 ± 0.8 µm, L/W ratio = 2.7.Asexual morph on SNA. Vegetative hyphae 1–8 µm diam,hyaline to pale brown, smooth-walled, septate, branched.Chlamydospores not observed. Conidiomata absent, conidiophoresformed directly on hyphae. Setae not observed. Conidiophoreshyaline to pale brown, smooth-walled, septate and branched, to50 µm long. Conidiogenous cells hyaline to pale brown, smoothwalled,cylindrical to ampulliform, often lacking a basal septumand continuous with the conidiophore, discrete phialides measure4–13 × 3–5 µm, opening 1–2 µm diam, collarette distinct, 1–2 µmlong, periclinal thickening distinct. Conidia hyaline, smooth-walled,aseptate, straight cylindrical with one end round and one endslightly acute or both ends round, (12–)12.5–17(–24) × (4–)4.5–5.5(–6) µm, mean ± SD = 14.7 ± 2.1 × 5.0 ± 0.7µm, L/W ratio =2.9. Appressoria single or in loose groups, medium to dark brown,smooth-walled, the outline mostly clavate, elliptical or ovate, theedge entire or undulate, rarely lobate, (5.5–)8–13(–17.5) × (4.5–)6–8(–9.5) µm, mean ± SD = 10.6 ± 2.4 × 7.0 ± 1.1 µm, L/W ratio= 1.5.Asexual morph on Anthriscus stem. Conidiomata acervular,conidiophores formed on a cushion of pale brown angular cells4–9 µm diam. Setae very few, pale brown, smooth-walled,3–4-septate, 50–80 µm long, base cylindrical, 3–3.5 µm diam, tip± rounded or ending with a conidiogenous locus. Conidiophorespale brown, smooth-walled, septate, branched, to 40 µm long.Conidiogenous cells pale brown, smooth-walled, cylindrical,sometimes polyphialides, 12–28 × 2–3.5 µm, opening 1–2 µmdiam, collarette 0.5–1.5 µm long, periclinal thickening distinct.Conidia hyaline, smooth-walled, aseptate, straight, very variable inshape, cylindrical, clavate, ellipsoidal or limoniform with one endround and one end slightly acute to truncate, (7.5–)10.5–17.5(–21)× (3.5–)4–5.5(–6) µm, mean ± SD = 14.1 ± 3.5 × 4.8 ± 0.6 µm,L/W ratio = 2.9.Culture characteristics: Colonies on SNA flat with entire margin,hyaline to pale cinnamon, on filter paper, Anthriscus stem andmedium covered with short floccose-felty pale olivaceous greyaerial mycelium, on Anthriscus stem covered with pale grey to blackstructures, reverse medium hyaline to pale cinnamon, filter paperpale cinnamon to olivaceous grey, growth rate 20–22.5 mm in 7 d(32.5–37.5 mm in 10 d). Colonies on OA flat with entire margin;surface honey, pale olivaceous grey, grey olivaceous to olivaceous,almost entirely covered with floccose-felty pale olivaceous greyaerial mycelium, reverse pale olivaceous grey, grey olivaceouswww.studiesinmycology.org95
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Studies in Mycology 73 (September 2
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Colletotrichum: complex species or
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Damm et al.Table 1. Strains of Coll
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Damm et al.Table 1. (Continued).Gen
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Damm et al.combined sequence datase
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Damm et al.Fig. 2. Colletotrichum a
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The Colletotrichum boninense specie
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available online at www.studiesinmy
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The Colletotrichum acutatum species
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The Colletotrichum acutatum species
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Weir et al.Fig. 17. Colletotrichum
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Weir et al.Fig. 18. Glomerella cing
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Weir et al.images under C. fructico
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Weir et al.Specimen examined: Thail
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Weir et al.Notes: First described f
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Weir et al.gossypii var. cephalospo
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Weir et al.to utilise either citrat
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Weir et al.(ex-epitype culture CBS
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Weir et al.of this species to C. ps
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Weir et al.Table 4. Performance of
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Weir et al.Crous PW, Gams W, Stalpe
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Weir et al.Simmonds JH (1968). Type
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Cannon et al.182
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Cannon et al.In rare instances, Col
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Cannon et al.Table 1. Summary of pr
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Cannon et al.Table 1. (Continued).P
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Cannon et al.Table 2. Colletotrichu
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Cannon et al.as did those for Colle
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Cannon et al.Table 3. Authentic seq
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Cannon et al.Table 3. (Continued).S
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Cannon et al.110.74 C. cuscutae IMI
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Cannon et al.Boninense cladeThe bon
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Cannon et al.C. lindemuthianum AB08
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Cannon et al.A further important ac
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Cannon et al.Hawksworth DL (2011).
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Cannon et al.Rodríguez-Guerra R, R
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Studies in MycologyStudies in Mycol
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CBS Biodiversity SeriesNo. 11:Taxon
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Studies in Mycology (ISSN 0166-0616
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