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

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Weir et al.to utilise either citrate or tartrate as a sole carbon source, to helpcharacterise isolates as C. kahawae. None of our C. kahawaesubsp. kahawae isolates were able to utilise either citrate or tartrate,whereas all of the C. kahawae subsp. ciggaro isolates were ableto utilise one or both of these carbon sources (Weir & Johnston2009). All of the C. kahawae subsp. kahawae isolates share a 22bp deletion in the glutamine synthetase gene, lacking in the C.kahawae subsp. ciggaro isolates. Note that one of the isolatesmetabolically and genetically typical C. kahawae subsp. kahawae(CBS 982.69) was reported by Gielink & Vermeulen (1983) to benon-pathogenic to coffee, but we have not independently checkedthis result.The isolates we accept as C. kahawae subsp. kahawae showtwo cultural types, one matching the description of Waller et al.(1993), slow growing, darkly pigmented cultures with conidiadeveloping mostly in the aerial mycelium. The second cultural typegrew even more slowly, had little or no pigmentation within the agar,and the colony surface was covered with numerous acervuli andorange conidial masses. Metabolically and genetically both culturaltypes were the same, and pathogenicity tests showed that the nonpigmentedisolates caused CBD (unpubl. data, D. Silva, Centro deInvestigação das Ferrugens do Cafeeiro). Rodriguez et al. (1991)reported further variation in cultural appearance amongst CBDcausing isolates.Waller et al. 1993 stated that C. kahawae was not known to formascospores. However, Gielink & Vermeulen (1983) observed theproduction of perithecia on coffee berries that had been inoculatedwith CBD-causing isolates, many months after inoculation and deathof the berries. At least one of the isolates that they cited with thisbiology, CBS 135.30, has the GS sequence typical of C. kahawaesubsp. kahawae. Vermeulen et al. (1984) grew cultures from theperithecia that developed on the previously inoculated berries, andfound that none were pathogenic to coffee. It is possible that theperithecia developing on inoculated berries reported by Gielink &Vermeulen (1983) were from other Colletotrichum spp. present onthe berries before they were inoculated, and represented speciesdistinct from C. kahawae subsp. kahawae. A similar situation hasbeen noted with some of our inoculations, where species presenton tissues prior to inoculation, either endophytic or latent, startedto sporulate on the dead tissue following inoculation (unpubl. data,B.S. Weir).Based on ITS sequences, most of the accessions in GenBankidentified as C. kahawae and isolated from coffee, match ourconcept of C. kahawae subsp. kahawae. There are two exceptions,AF534468 (from Malawi) and AY376540 (STE-U 5295 = IMI 319424= CBS 112985, from Kenya). The Kenyan isolate was cited as C.kahawae in Lubbe et al. (2004). Based on the ITS sequences, andthe TUB2 sequence from isolate STE-U 5295 (AY376588), theseisolates represent C. siamense.Specimens examined: Angola, Ganada, on Coffea arabica berry, coll. J.N.M. Pedro16/65, 2 Jun. 1965 (IMI 310524 = CBS 982.69 = ICMP 17915). Cameroon, onCoffea arabica (IMI 361501 = ICMP 17905). Kenya, Ruiru, Kakuzi Estate, on Coffeaarabica young shoots, coll. D.M. Masaba 22/87, 29 Jan. 1987 (ex-holotype cultureof C. kahawae – IMI 319418 = ICMP 17816); on Coffea sp., coll. E.C. Edwards,May 1930 (CBS 135.30 = ICMP 17982). Malawi, on Coffea arabica (IMI 301220 =ICMP 17811).* Colletotrichum kahawae subsp. ciggaro B. Weir & P.R.Johnst., subsp. nov. MycoBank MB563758. Figs 26, 27.= Glomerella cingulata var. migrans Wollenw., Z. Parasitenk. (Berlin) 14: 262.1949.= Glomerella rufomaculans var. vaccinii Shear, Bull. Torrey Bot. Club. 34: 314.1907.Etymology: Based on the title of the Jim Jarmusch movie “Coffeeand Cigarettes”, referring to the close genetic relationship betweenC. kahawae subsp. ciggaro and the coffee pathogen C. kahawaesubsp. kahawae; ciggaro is Portuguese for cigarette.Holotype: Australia, on Olea europaea, coll. V. Sergeeva UWS124,1989, PDD 102232; ex-type culture ICMP 18539.Colonies grown from single conidia on Difco PDA 75–85 mm diamafter 10 d for most isolates, the ex-holotype culture of G. cingulatavar. migrans 48–49 mm diam. Aerial mycelium cottony, grey, dense,or in some isolates with dark stromatic masses and associatedorange conidial ooze showing through mycelium from agarsurface; in reverse agar with pinkish-orange pigments (6B4–7B4),irregular scattered black spots, and variable levels of developmentof overlying dark grey to green-grey pigments (4C2–5D4), thesesometimes in discrete sectors. See notes below about a divergentgrowth form single ascospore cultures from perithecia in culture.Conidia form on dark-based acervuli, (12–)16–19.5(–29) × (4.5–)5(–8) µm (av. 17.8 × 5.1 µm, n = 214), cylindric, straight, apexrounded, often tapering slightly towards the base. Appressoriatypically cylindric to fusoid in shape, deeply lobed. Perithecianumerous, forming tightly packed clumps, individual peritheciaglobose, small, about 250 µm diam, with a short ostiolar neck. Asci55–100 × 10–12 µm, 8–spored. Ascospores (13.5–)17.5–20(–24)× (4–)4.5–5(–6.5) µm (av. 18.8 × 4.8 µm, n = 121), gently curved,tapering to quite narrow, rounded ends, widest point usuallytowards one end of the spore.Geographic distribution and host range: Known from Australia,Germany, New Zealand, and South Africa. Both host and geographicrange of the isolates we accept in C. kahawae subsp. ciggaroare broad. Genetic identification: ACT, CAL, CHS-1, GAPDH,TUB2, SOD2, and ITS sequences match those from C. kahawaesubsp. kahawae. The two subspecies can be distinguished by GSsequences. Collectively, the two subspecies can be distinguishedfrom all other species using ITS sequences alone.Notes: The authentic isolate of G. cingulata var. migrans (CBS 237.49)differed from all other isolates we accept in C. kahawae subsp.ciggaro by its slower growth rate. Wollenweber & Hochapfel (1949)distinguished Glomerella cingulata var. migrans from G. cingulata var.cingulata on the basis of pathogenicity (G. cingulata var. migrans waspathogenic to Hypericum and not to apple) and because of its slightlylonger ascospores and shorter conidia — ascospores average 21 ×4.2 µm versus 18 × 4.6 µm, conidia average 14 × 5.2 µm versus 18 ×5 µm (Wollenweber & Hochapfel 1949). We were unable to produceascospores from CBS 237.49, the conidia were similar in size to thatreported by Wollenweber & Hochapfel (1949), averaging 16.6 × 5.3µm. However, the average ascospore and conidial lengths of our C.kahawae subsp. ciggaro isolates varied across the range cited byWollenweber & Hochapfel (1949) for both G. cingulata var. cingulataand G. cingulata var. migrans, the average ascospore length fromindividual isolates ranging from 16.6 to 20 µm, the average conidiallength ranging from 14.9 to 21.2 µm.Glomerella rufomaculans var. vaccinii was described by Shear(1907) for a fungus isolated from cranberry that was morphologicallyidentical to isolates from apple and other hosts but which appearedto be biologically distinct (Shear 1907, Shear & Wood 1913). Aputatively authentic isolate of this species, deposited by Shearin CBS in 1922, matches C. kahawae subsp. ciggaro genetically.Polashock et al. (2009) discussed the diversity of Colletotrichum158
The Colletotrichum gloeosporioides species complexFig. 26. Colletotrichum kahawae subsp. ciggaro. A. ICMP 12952. B, D. ICMP 17932 (ex CBS 112984). E, H. ICMP 17931 (ex IMI 359911). C, F. ICMP 18539 – ex-holotypeculture. G. ICMP 18531. A–B. Asci and ascospores. C–D. Appressoria. E. Setae. F–H. Conidia. Scale bars A, E = 20 µm. Scale bar of A applies to A–D, F–H.spp. associated with North American cranberry fruit rots, reporting aclose match between their isolates and C. kahawae. Incorporationof their ITS sequences into our alignment confirms this. Whether ornot there is a genetically distinct, cranberry specialised taxon withinC. kahawae requires additional genes to be sequenced from thecranberry-associated isolates.Colletotrichum kahawae subsp. ciggaro was referred to as C.gloeosporioides Group B by Johnston & Jones (1997) and Johnstonwww.studiesinmycology.org159
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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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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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Damm et al.Fig. 25. Colletotrichum
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Damm et al.to olivaceous grey, filt
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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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