mycological research 110 (2006) 1395–1408

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Species of on Agavaceae

David F. FARRa, M. Catherine AIMEa, Amy Y. ROSSMANa,*, Mary E. PALMb aSystematic Botany & Mycology Laboratory, Agricultural Research Service, United States Department of Agriculture, Rm 304, B011A, Beltsville, Maryland, USA bSystematic Botany & Mycology Laboratory, Animal and Plant Health Inspection Service, United States Department of Agriculture, Rm 304, B011A, Beltsville, Maryland, USA article info abstract

Article history: Species of Colletotrichum cause diseases on a wide range of hosts, frequently infecting Received 22 December 2005 plants in the Agavaceae (monocotyledons: Liliales). Three species of Colletotrichum restricted Received in revised form to the Agavaceae were detected through morphological studies of specimens and molecular 30 June 2006 sequence analyses of the LSU of the nu-rDNA and the ITS region of the nu-rDNA from cul- Accepted 1 September 2006 tures. Colletotrichum agaves on Agave is fully described and illustrated. Colletotrichum dracae- Published online 28 November 2006 nophilum is described as a new species for isolates having long conidia and occurring on Corresponding Editor: Dracaena sanderiana from China. Colletotrichum phormii and Glomerella phormii are deter- Brenda Wingfield mined to be the correct scientific names for the asexual and sexual states, respectively, of a species commonly referred to as C. rhodocyclum and G. phacidiomorpha occurring mainly Keywords: on Phormium. In addition, C. gloeosporioides and C. boninense were isolated from plants in the Anthracnose Agavaceae. All species of Colletotrichum described on Agavaceae were evaluated based on Molecular phylogeny type specimens. A key to the five species of Colletotrichum on Agavaceae is included. This pa- Plant pathology per includes one new species, Colletotrichum dracaenophilum, and three new combinations, Colletotrichum phormii, Glomerella phormii, and Phaeosphaeriopsis phacidiomorpha. Published by Elsevier Ltd on behalf of The British Mycological Society.

Introduction encountered as of tropical woody plants (Arnold et al. 2003; Lu et al. 2004; Promputtha et al. 2005) as well as her- Members of the genus Colletotrichum cause diseases on baceous and monocotyledonous plants (Photita et al. 2004, a number of plants. These diseases, often referred to 2005). The taxonomy and phylogenetics of most species of Col- as anthracnose, include black spot of and key letotrichum have not been studied, which limits accurate lime anthracnose (C. acutatum), anthracnose of knowledge of the distribution and biology of these species, as (C. coccodes), red stalk rot (C. graminicola), coffee well as the ability to identify individual taxa. berry disease (C. kawahae), anthracnose of beans (C. lindemu- Historically, it was assumed that species of Colletotrichum thianum) and many others (Holliday 2000). According to Sutton were host-specific. New species were described when this (1992:1) species of Colletotrichum are especially important as the was encountered on a new host plant genus resulting cause of preharvest and postharvest problems in the tropics ‘as in more than 660 names published in Colletotrichum (Index latent or quiescent infections’. Some plant pathogenic species Fungorum http://www.indexfungorum.org/Names/Names. of Colletotrichum are of quarantine significance because they asp). About 600 of these were considered synonyms of could be introduced into countries where the disease does C. gloeosporioides by Arx (1957) based on his morphological not occur. Species of Colletotrichum are also frequently examination of type specimens. In later studies some of

* Corresponding author. E-mail address: [email protected] 0953-7562/$ – see front matter Published by Elsevier Ltd on behalf of The British Mycological Society. doi:10.1016/j.mycres.2006.09.001 1396 D. F. Farr et al.

these synonyms have proven to be distinct species. A useful key for the identification of species of Colletotrichum was Materials and methods published by Sutton (1980) who distinguished 21 species and species-groups based primarily on characteristics of the Morphological/cultural characterization conidia, appressoria, setae, sclerotia, and colonies produced under standardized conditions combined with plant host. Cultures were obtained from acervuli and perithecia in leaves Later Sutton (1992) listed 39 accepted species in Colletotrichum of specimens intercepted at ports of entry in the US. A small each with a short description but no key. At present about number of conidia were transferred from the host substrata 40 species are accepted in the genus Colletotrichum (Kirk to sterile water in a hanging drop slide, mixed, and spread et al. 2001), of which 25 are represented in GenBank. How- on a plate of corn meal dextrose agar (Difco, Detroit, MI) plus ever, most species of Colletotrichum have not been studied antibiotics (CMD-A, 0.2 % neomycin and 0.2 % streptomycin). in culture or using molecular, biochemical or genetic tech- After incubation overnight at room temperature, single or niques. Neither of the two previously described species on multiple germinating conidia were transferred to new CMD-A Agavaceae included in this paper was accepted by Sutton and dextrose agar (PDA, Difco, Detroit, MI) plates for (1992). processing. Additional cultures were obtained from the Cen- Recent molecular systematics research on the genus Col- traalbureau voor Schimmelcultures (CBS), Utrecht. letotrichum suggests that some taxa are host specific while For microscopic examination infected plant material was others are not. Using multiple gene phylogenies, RFLPs, rehydrated and mounted in 3 % potassium hydroxide. Sec- and mating tests, some easily recognizable morphological tions of acervuli ca 10 mm thick were obtained with a freezing species have been shown to consist of genetically distinct microtome and mounted in lactic acid with cotton blue. Ob- groups. For example, within C. acutatum seven distinct mo- servations of microscopic features were made using a Zeiss lecular groups could be identified (Guerber et al. 2003). Mat- Axioplan 2 microscope with bright-field and fluorescence illu- ing compatibility was determined within one clade that mination. Calcofluor was used as the fluorescent dye. Photo- included isolates from a wide range of hosts and geographic graphs and measurements of microscopic features were origins but also occurred between two phylogenetically dis- taken using an Olympus Q5 digital camera (Olympus, Tokyo) tinct clades, one of which included only isolates from Aus- and ImagePro software (Media Cybernetics, Silver Spring, MD). tralia and New Zealand that cause fruit rots and terminal The description of cultural characteristics was based on crook disease of pine seedlings. Within the C. acutatum com- isolates grown at 25 C, under NUV plus fluorescent light on plex, a new species, C. lupini, with one variety C. lupini var. PDA for 6 d. Colours were described using terms in Raynor setosum, has recently been recognized (Nirenberg et al. (1970). Sterilized stems ( sativa) on water agar 2002) that occurs only on . This species was originally were inoculated with each isolate and maintained under the considered to be C. gloeosporioides (Elmer et al. 2001). Other same conditions listed above in order to obtain dried cultures species are being segregated within the C. gloeosporioides as herbarium specimens. Slide cultures using PDA were made complex, such as C. kahawae, cause of coffee berry disease for each isolate (Malloch 1981) and examined for the develop- (Lubbe et al. 2004). Based on isolates from diseased leaves ment of appressoria. of eight different plant species in Japan, Colletotrichum Parent specimens from which cultures were derived, as boninense (Moriwaki et al. 2003) was described and distin- well as dried cultures were deposited at the US National Fun- guished both morphologically and by sequences of the ITS gus Collections (BPI), Beltsville, Maryland. Living cultures were region of the nu-rDNA repeat. Two of the eight groups of deposited at the CBS. Colletotrichum isolated as endophytes from tropical were considered to be related to C. boninense (Lu et al. DNA isolation, sequencing and analyses 2004). Thus, some species of Colletotrichum may function as both plant pathogens and endophytes (Photita et al. 2004, Provenance of cultures used for sequencing analyses is listed 2005). in Table 1. Mycelium for DNA extractions was cut with a sterile Plants belonging to the Agavaceae (monocotyledons: Lil- scalpel from actively growing cultures on PDA and extracted iales) intercepted at US ports of entry are frequently infected with the UltraClean Plant DNA Isolation Kit as per the manu- with species of Colletotrichum. At present the United States is facturer’s instructions (MoBio Laboratories, Inc., Solana prohibiting the entry of Phormium tenax (New Zealand flax) Beach, CA). Individual genes were amplified by PCR in 25 ml re- infected with a long-spored species of Colletotrichum that has actions with 12.5 ml of PCR Master Mix (Promega, Madison, WI), been called C. rhodocyclum. Additional species of Colletotrichum 1.25 ml each of 10 mM primers and 10 ml diluted (10- to 100-fold) having conidia greater than 20 mm have been encountered on DNA template using the following primers: LSU nu-rDNA LR0R living plants of Dracaena sanderiana (lucky bamboo) from and LR7 (Vilgalys & Hester 1990); ITS rDNA ITS1-F (Gardes & China. Two-gene molecular analyses of all available Colletotri- Bruns 1993) and ITS4 (White et al. 1990). Standard cycling pa- chum species were undertaken in this study in order to clarify rameters with a 55 C annealing temperature were used for the identity and number of species of Colletotrichum known on both gene regions. The PCR products were cleaned with Mon- the Agavaceae. Five species were isolated from plants in the tage PCR Centrifugal Filter Devices (Millipore Corp., Billerica, Agavaceae, including one new species, C. dracaenophilum. MA) according to the manufacturer’s protocol and sequenced Three of these species are described, illustrated and dis- as described in Rossman et al. (2004). Sequences have been de- cussed, and a key to the five species of Colletotrichum on Agava- posited in GenBank (http://www.ncbi.nlm.nih.gov) and are ceae is provided. listed in Table 1. Colletotrichum on Agavaceae 1397

Table 1 – Isolates of Colletotrichum newly sequenced for this study with reference numbers, country, plant host genus, and GenBank numbers Scientific name Culture/voucher specimen no. Country Host GenBank GenBank no. LSU no. ITS

Colletotrichum 1ATCC MYA-662 USA Arkansas Malus DQ286122 DQ286121 acutatum ATCC MYA-664 USA Arkansas Malus DQ286223 DQ286123 MEP 1323, CBS 119292 New Zealand Vaccinium DQ286125 DQ286124 MEP 1325, CBS 119186/BPI 1107284 New Zealand Vaccinium DQ286127 DQ286126 MEP 1322, CBS 119293/LA 093243 New Zealand Vaccinium DQ286129 DQ286128 ATCC 56816 d type culture Australia Carica DQ286133 DQ286132 MEP 1534, CBS 119294/BPI 871836 Mexico Leucaena DQ286131 DQ286130 C. agaves CBS 318.79 (as C. crassipes)/BPI 871832 Netherlands Agave DQ286220 DQ286219 AR 3920, CBS 118190/BPI 871831 Mexico Agave DQ286222 DQ286221 C. boninense MEP 1554, CBS 118401/BPI 871492 China Pachira DQ286161 DQ286160 AR 3649/BPI 749129 China Pachira DQ286163 DQ286162 AR 3739, CBS 119185/BPI 871835 Brazil Hippeastrum DQ286165 DQ286164 CBS 241.78 The Netherlands Hippeastrum DQ286167 DQ286166 AR 3658, CBS 118193/BPI 748487 China Dracaena DQ286169 DQ286168 AR 3751, CBS 118774/BPI 871833 China Dracaena DQ286171 DQ286170 CBS 102667 New Zealand Passiflora DQ286173 DQ286172 C. capsici AR 4028, CBS 119189 USA Maryland Phaseolus lunatum DQ286157 DQ286156 AR 4048, CBS 119295 Costa Rica Jatropha DQ286159 DQ286158 C. crassipes CBS 159.75 India Ex air and DQ286206 DQ286205 stored grain C. dematium AR 3563, CBS 119444/Houston 05763 Mexico Lirope DQ286155 DQ286154 C. dracaenophilum MEP 1532, CBS 118199/BPI 871498 Holotype China Dracaena DQ286210 DQ286209 MEP 1537, CBS 119360/BPI 871491 China Dracaena DQ286208 DQ286207 MEP 1518, CBS 118200/BPI 842098 China Dracaena DQ286212 DQ286211 C. gloeosporioides AR 3385 Puerto Rico Cajanus DQ286175 DQ286174 GJS 01-199/BPI 842147 Cameroon Theobroma DQ286177 DQ286176 AR 4040, CBS 119195 Panama Theobroma DQ286195 DQ286194 MEP 1525, CBS 119201 Jamaica orchid DQ286179 DQ286178 AR 4041 Panama Theobroma DQ286181 DQ286180 5101 Panama Theobroma DQ286183 DQ286182 MCA 2499, CBS 119290 Hungary Unknown DQ286197 DQ286196 MCA 2498, CBS 119296 Unknown Glycine DQ286199 DQ286198 AR 4042, CBS 119298 Panama Theobroma DQ286185 DQ286184 AR 2802, 119299/BPI 871839 USA Georgia Pueraria DQ286187 DQ286186 AR 2801, CBS 119205/BPI 871838 USA Georgia Pueraria DQ286191 DQ286190 AR 2800, CBS 119203 USA Georgia Pueraria DQ286189 DQ286188 AR 2799, CBS 119204/BPI 871837 USA Georgia Pueraria DQ286193 DQ286192 MCA 2503 USA Maryland (greenhouse) Theobroma DQ286204 DQ286203 C. lupini AR 2820/BPI 863581 USA: Utah Lupinus DQ286118 DQ286117 AR 2826/BPI 871840 USA: New Hampshire Lupinus DQ286120 DQ286119 C. phormii MEP 1334, CBS 118201/BPI 802177 New Zealand Phormium DQ286141 DQ286140 CBS 198.35 cultures ex Kinghorn England Phormium DQ286145 DQ286144 CBS 199.35 cultures ex Kinghorn England Phormium DQ286143 DQ286142 AR 3410, CBS 118195/SF 051697 South Africa Phormium DQ286139 DQ286138 AR 3389, CBS 118197/BPI 746430 New Zealand Phormium DQ286135 DQ286134 AR 3546, CBS 118194 Germany Phormium DQ286137 DQ286136 AR 3787, CBS 118191/BPI 871493 South Africa Phormium DQ286147 DQ286146 Colletotrichum sp. MEP 1545, CBS 119292/Miami 223820 Panama Cygnoches DQ286149 DQ286148 AR 4031, CBS 114801/BPI 843682 Greece Crupina AY539807 AY539806 MEP 1530, CBS 119206/BPI 842096 Italy Ruscus DQ286151 DQ286150 AR 3749/BPI 871027 Thailand Cattleya DQ286214 DQ286213 AR 3750/BPI 871023 Thailand Dendrobium DQ286216 DQ286215 MCA 2773, CBS 119297 Costa Rica Theobroma DQ286218 DQ286217 C. trichellum MEP 1535, CBS 118198/BPI 871496 Guatemala Hedera DQ286153 DQ286152 AR 3788, CBS 118192/BPI 871497 Indonesia Dracaena DQ2862001 DQ286200 FAU 552, CBS 120005/BPI 747977 USA: Florida Fragaria DQ286225 DQ286224 FAU 553, CBS 114054/BPI 747978 USA: Florida Fragaria AF543786 DQ286202

1 AR, Amy Rossman; ATCC, American Type Culture Collection; BPI, US National Fungus Collections; CBS, Centraal Bureau voor Schimmelcul- tures; FAU, Francis A. Uecker; GJS, Gary J. Samuels; MCA, M. Catherine Aime; MEP, Mary E. Palm. 1398 D. F. Farr et al.

Sequencing reactions were edited and contiguous se- Phylogenetic analyses quences were assembled in Sequencher v.4.1.4 (Gene Codes Corp., Ann Arbor, MI). Sequences obtained were blasted in Fifty-five new sequences from isolates of Colletotrichum/Glom- GenBank to confirm identity as Colletotrichum spp. Plecto- erella were analysed along with 17 sequences selected from sphaerella cucumerina was chosen as an outgroup as this GenBank. A comprehensive dataset was assembled that in- fungus represents the closest known relative to Glomerella/ cludes both ITS and LSU sequences from 27 of the estimated Colletotrichum (Huhndorf et al. 2004). Sequence alignments 40 species of Colletotrichum (Kirk et al. 2001). Of the 1185 char- were constructed by eye in Se-Al v2.0a11 (http://evolve. acters included, 970 were constant, 111 variable characters zoo.ox.ac.uk) for each gene and then concatenated in PAUP were parsimony uninformative, and 104 variable characters v.4.0b10 (Swofford 2002). Sequences were selected from Gen- were parsimony informative. Four equally parsimonious trees Bank for those Colletotrichum and outgroup taxa for which of length 402 were found by MP; CI ¼ 0.61; RI ¼ 0.87. Both MP both ITS and LSU sequences were available from the and ML methods uncovered the same major lineages (Fig 1). same isolate. These were: Colletotrichum sp. AY539806 and AY539807, C. circinans AJ301955, C. coccodes AB105970 and AB06153, C. dematium AJ301954, C. destructivum AF320562 and AB106148, C. fuscum AB105966 and AB106161, C. higgin- Taxonomy sianum AB105957 and AB105956, C. lindemuthianum AJ301958, C. linicola AB057437 and AB106150, C. sublineolum Colletotrichum agaves Cavara, Fungi Longobardiae Exsiccati No. AJ301978, C. trifolii AJ301941, C. truncatum AJ301985, Glomer- 100 (1892). ella cingulata AF543786, G. glycines AB057435 and AB106160, Fig 2 G. lagenerium AJ301965, and Plectosphaerella cucumerina Leaf lesions circular, chlorotic, with or without a brown mar- AF176952 and AF176953. Data analyses followed Rossman gin, usually 2–4 cm diam, up to 6 cm diam or more, several on et al. (2004). The dataset consisted of 70 isolates aligned one leaf, occasionally becoming confluent, eventually causing across 1272 bp. A total of 86 bp were excluded from analyses death of plant tissue that becomes purplish to dark brown. due to ambiguities in alignment. MP analyses were Acervuli epiphyllous, scattered or forming concentric rings, conducted in PAUP as heuristic searches with 100 random distinct to confluent, developing beneath epidermis, becom- addition replicates and bisection-reconnection branch ing erumpent with age, often surrounded by remnants of epi- swapping. Support for the branching topologies was dermis, globose to oblong, in longitudinal section of hyaline to evaluated by BS analysis derived from 1000 replicates with dark brown cells forming textura angularis, 0.10–1 mm wide, ten random addition replicates each. ML analyses were con- 200–400 mm tall, brown to black, producing masses of pale or- ducted by the quartet puzzling method (Strimmer & von ange conidia. Setae numerous, projecting beyond hymenial Haeseler 1996) in PAUP with 10 K puzzling steps; transition: surface, brown to black, thick-walled, septate, straight to ir- transversion ratio ¼ 2. regularly crooked, tapering to acute or rounded apices, 60– 211 5–8 mm. Conidiophores dark brown to hyaline arising from upper cells of acervuli. Conidiogenous cells hyaline, Results enteroblastic, 18.5–24 3.5–7 mm, cylindric, periclinal thicken- ings moderate, channel narrow to broad. Conidia hyaline, Five different species of Colletotrichum were isolated from (17.5–) 19.0–30.5 (–33) 5–8 (–9.5) mm (mean ¼ 23.5 6.5 mm, plant hosts in the Agavaceae. Three of these species appear sd length ¼ 3 mm, width ¼ 0.5 mm, n ¼ 146), aseptate, cylindri- to occur only on specific genera in the Agavaceae, viz. C. agaves cal or expanding to a broadly rounded apex, straight or slightly on Agave, C. dracaenophilum on Dracaena, and C. phormii on curved, with one or two guttules. Phormium, as indicated by the phylogenetic analyses (see be- Colonies 3.7–4.2 cm diam, rosy buff to sepia, peach at mar- low in Results). These three species are described and illus- gin, margin smooth, broadly wavy, reverse rosy buff to peach, trated in the taxonomy section of this paper. Two additional profuse sporulation on agar and alfalfa twigs, acervuli similar species having a wide host range were identified from Dra- to above, open acervuli each with numerous black setae. caena. Two isolates of Colletotrichum from Dracaena were iden- Appressoria not seen. All other characters as on natural tified as C. boninense based on both molecular and substrata. morphological characteristics. The following specimens of Host range: On numerous species of Agave as well as the C. boninense were examined: China: on living leaves of closely related genus Furcraea, including F. macrophylla Baker Dracaena sanderiana, APHIS interception Hoboken New Jersey, (wild sisal). 30 Apr. 2001, P. Milicia, det. M. Feinstein, isol. AR 3658 ¼ CBS Geographic distribution: Known from warm temperate and 118193 (BPI 748487); 25 Oct. 2001, P. Milicia & M. Feinstein, isol. tropical regions including southern Europe, the Neotropics, AR 3751 ¼ CBS 118774 (BPI 871833). One isolate Cyprus (Georghiou & Papadopoulos 1957); Africa (Kenya in was obtained from a specimen of Glomerella cingulata on Nattrass 1961); and the US (FL, MO in greenhouses, MS in Par- Dracaena that grouped with other sequences of G. cingulata in- ris 1959). Although no specimens were located and examined, cluding the homothallic isolates studied by Uecker (1994). The C. agaves has also been reported from Asia, including China following specimen of G. cingulata was examined: Indonesia: (Tai 1979), Korea (Cho & Shin 2004), the Philippines (Teodoro on Dracaena, APHIS interception Miami Florida 223164, 4 Mar. 1937) and Taiwan (Anonymous 1979). 2002, H. Ruiz, det. A.Y. Rossman, isol. from AR Type specimens examined: Italy: Pavia: Ticinum, in botanical 3788 ¼ CBS 118192 (BPI 871497). garden, on decaying leaves of cultivated Agave americana, Colletotrichum on Agavaceae 1399

91/53 Colletotrichum lupini AR2820 Colletotrichum lupini AR2826 64/86 MYA-662 Colletotrichum acutatum MYA-664 Colletotrichum acutatum MEP1323 Colletotrichum acutatum MEP1325 Colletotrichum acutatum MEP1322 Colletotrichum acutatum ATCC56816 Colletotrichum phormii AR3389 53/56 Colletotrichum phormii AR3410 99/91 Colletotrichum phormii AR3546 Colletotrichum phormii AR3787 Colletotrichum phormii CBS198.35 96/90 Colletotrichum phormii CBS199.35 Colletotrichum phormii MEP1334 Colletotrichum acutatum MEP1534 Colletotrichum sp. MEP1545 Colletotrichum sp. AR4031 Colletotrichum higginsianum Colletotrichum destructivum Colletotrichum fuscum Colletotrichum linicola 67/93 Glomerella glycines AR3563 Colletotrichum dematium AJ301954 100/94 AR4028 Colletotrichum capsici AR4048 55/69 Colletotrichum sp. MEP1530 63/72 Colletotrichum trichellum Colletotrichum circinans Colletotrichum sublineolum 78/76 Colletotrichum boninense MEP1554 Colletotrichum boninense AR3649 80/59 94/96 Colletotrichum boninense AR3739 Colletotrichum boninense CBS241.78 Colletotrichum boninense AR3658 Colletotrichum boninense AR3751 Colletotrichum boninense CBS102667 98/97 Colletotrichum agaves CBS318.79 Colletotrichum agaves AR3920 Colletotrichum gloeosporoides AR3385 Colletotrichum sp. GJS01.199 Colletotrichum gloeosporoides AR4040 Colletotrichum sp. MEP1525 Glomerella cingulata AR4041 65/92 Colletotrichum gloeosporoides 5101 Colletotrichum gloeosporoides MCA2499 Colletotrichum gloeosporoides MCA2498 Glomerella cingulata AR3788 Glomerella cingulata FAU552 Glomerella cingulata FAU553 100/98 Colletotrichum gloeosporoides AR4042 67/99 Colletotrichum gloeosporoides AR2802 Colletotrichum gloeosporioides AR2801 Colletotrichum gloeosporioides AR2800 Colletotrichum gloeosporoides AR2799

82/85 Colletotrichum gloeosporoides MCA2503 Colletotrichum dracaenophilum MEP1537 95/58 Colletotrichum dracaenophilum MEP1518 Colletotrichum dracaenophilum MEP1532 99/88 Glomerella sp. AR3749 98/77 Colletotrichum sp. AR3750 Colletotrichum sp. MCA2773 Glomerella lagenerium 95/98 Colletotrichum lindemuthianum Plectosphaerella cucumerina 5 changes

Fig 1 – MP tree derived from combined ITS and LSU sequence matrix. Nodes with moderate support by bootstrapping (50–70 %) and quartet puzzling (50–70 %) are of medium thickness. Nodes with both strong bootstrapping support (>70 %) and quartet puzzling reliability (>70 %) are indicated by heavily blackened branches. Support values are given above supported branches: the number before the slash represents bootstrapping support values from MP analyses; the number after the slash represents quartet puzzling reliability scores for that node. Isolates from Agavaceae are indicated in bold type. 1400 D. F. Farr et al.

Fig 2 – Colletotrichum agaves. A. Conidiomata on natural substrata. BPI 397108. B. Section of conidiomata showing conidio- phores and setae in culture. CBS 118190. C. Section of conidiomata showing conidiophores, setae lacking on natural sub- strata. BPI 397108. D. Conidiophores and setae variable in length in culture. CBS 118190. 6–7. Conidia. E. Produced in culture. CBS 118190. F. Produced on natural substrata. BPI 397108. Bars: A [ 200 mm, B–D [ 100 mm, E–F [ 10 mm.

Cavara (BPI specimen in bound exsiccati – lectotypus hic desig- A.Y. Rossman AR 3920, CBS 118190 (BPI 871831); on Agave americana, natus; additional isolectotype specimens BPI 397134 and BPI APHIS interception Laredo, Texas 15693, 31 Jan. 1982, isol. & det. A. 397135). Rossman AR 1686 (BPI 396815); on Agave sp., APHIS interception in New York City, 12 Mar. 1923, J.W. O’Brien, det. J.A. Stevenson (BPI Additional specimens examined: Cuba: Cardenas,onAgave rigida d Baarn Agave var. sisalana, 27 Jul. 1903, J.H. Foster (BPI 397153); Havana:onAgave 397125). The Netherlands: , causing leaf spot on americana H.A. van der Aa rigida, Mar. 1926, Mr. Moreland & Mr. Nicaise 708, det. W.W. Diehl (BPI , Mar. 1979, isol. , No. 6863, CBS 318.79 ini- tially identified as C. crassipes (BPI 871832 dried culture). d USA: 397152); intercepted in Philadelphia, Pennsylvania, on diseased Florida Agave C.L. Shear F. Petrak leaves of Agave sp., 8 Jan. 1934, M. Kislink, det. J.A. Stevenson (BPI : Winter Park, on sp., Jan. 1946, , det. Agave C.L. Shear 397121). d El Salvador: Lake Ilopango Road,onAgave americana, (BPI 397115); Altamonte Spa, on sp., 24 Mar. 1938, , det. F. Petrak (BPI 397117); Missouri, St Louis, in greenhouse of the 25 Jul. 1943, F.L. Wellman 300, det. J.A. Stevenson (BPI 397130); San Missouri Botanical Garden, on Agave sp., G.G. Hedgcock (BPI Miguel: Santa Tecla, on Agave letonae, 16 Mar. 1949, B.S. Crandall, Agave americana G.G. Hedgcock det. J.A. Stevenson (BPI 397148). d Haiti: Port au Prince,onAgave 397108); on , 16 Oct. 1905, 466 (BPI Agave ‘expatriata G.G. Hedgcock sp. (sisal), 5 Sep. 1956, Felix Pierre-Louis (BPI 397112). d Italy: LaMos- 397136); on ?’, 16 Oct. 1909, 469 (BPI 397143). tala,onAgave sp., 1905, W. Trelease (BPI 397109). d Jamaica:on Agave sisalana Perrine, Nov. 1920, coll. S.F. Ashby (BPI 39711). d Mexico:onAgave striata, APHIS Interception Los Indios, Texas Notes: Colletotrichum agaves causes a foliar disease of agave 038140, 10 Jun. 2002, O. Mireles, det. N. Guerrero & M.E. Palm, isol. plants as first documented by Hedgcock (1905) who reported Colletotrichum on Agavaceae 1401

plants dying from this fungus in greenhouses at the Missouri 3–6 septate. Conidiogenous cells hyaline, enteroblastic, Botanical Garden, St Louis. This disease was also discussed by 18–32 4–8 mm, periclinal thickenings moderate, channel nar- Barthelet (1942) in Italy. Petrak (1942) reported it on plants row to broad. Conidia (20–)22–34(–38) (5.5–)6.5–9.5(–10) mm growing outdoors in Florida. (mean ¼ 29 8.5, sd length ¼ 3.5 mm, width ¼ 0.9 mm, n ¼ 190), C. agaves is easily distinguished from other species on broadly clavate to cylindrical, frequently slightly curved, hya- Agavaceae by the numerous black setae that develop line, guttulate. throughout the conidiogenous region in acervuli. Acervuli Colonies 6–6.7 cm diam, pale pink except dark where often develop in concentric rings in necrotic areas on the sporulating, margin pale pink, reverse speckled from profuse upper surface of living leaves of Agave. Acervuli of Colletotri- sporulation, sparse aerial mycelium, rosy buff to saffron in chum from Agave developing on alfalfa stems in culture ap- centre, rosy buff to saffron in reverse, except for presence of pear to be morphologically the same as C. agaves present dark acervuli, paler at even margin; immature, white acervuli on the type and other specimens. In addition after 6 d on developing at margin. Acervuli on alfalfa stems the same as on PDA, the growth rate of C. agaves (3.7–4.2 cm) is faster natural substrata except slightly smaller, 140–420 mmin than the slow-growing C. phormii (2.5–3.4 cm) but slower longest dimension, separate, and superficial. than C. dracaenophilum (6–6.7 cm). Arx (1957) listed C. agaves Host range: On stems of Dracaena sanderiana and Dracaena sp. among the several hundred synonyms of C. gloeosporioides Geographic distribution: Known only from China. while Hughes (1958) considered C. agaves a synonym of Additional specimens examined: China: on stems of Dracaena sand- C. coccodes. The molecular data for C. agaves show this eriana, APHIS interception Hoboken, New Jersey 104782, 12 Feb. M. Feinstein M.E. Palm species to be a distinct taxon, separate from both C. gloeo- 2003, det. , isol. (MEP 1537, CBS 119360, BPI 871491); APHIS interception at San Francisco, California 052192, sporioides and C. coccodes (Fig 1). No teleomorph is known 29 Jan. 2002, det. M. D. Petrillo, isol. M.E. Palm (MEP 1518, CBS for this species. 118200, BPI 842098). Notes: C. dracaenophilum is distinguished from all other spe- cies of Colletotrichum known from Agavaceae by the long coni- Excluded and doubtful names of Colletotrichum on Agave dia that average more than 28 mm long. In culture, C. dracaenophilum is the fastest growing of the three long-spored Gloeosporium agaves Syd. & P. Syd., Ann. Mycol. 5: 362 (1907). No species of Colletotrichum on Agavaceae. Although Dracaena conidia were found on the immature type specimen. This sanderiana is native to western Africa, this host is now culti- name was considered to be a synonym of Colletotrichum cras- vated throughout the world especially as ‘lucky bamboo’ in sipes by Arx (1957). small containers. Specimen examined: Brazil: Sao Paulo: Campinas, on Agave sp., Apr. 1898, F. Noark, comm. P. Sydow, ex herb Pazschke (B). Gloeosporium macropus was considered a possible synonym Excluded names of Colletotrichum on Dracaena of C. agaves by Hedgcock (1905) but this species is reported only on hosts other than Agave. The type specimen was not Colletotrichum dracaenae Allesch., in Rabenhorst, Rabenh. examined. Krypt.-Fl. 7: 560 (1902). The type specimen of this taxon was examined. The conidia Colletotrichum dracaenophilum D.F. Farr & M.E. Palm, were originally described as 14–18 5–7 mm, which agrees sp. nov. with our examination of the specimen in which the conidia Fig 3A–F were 14.5–18.5 5–8.5 mm (mean ¼ 16 6.5 mm, sd length ¼ MycoBank no.: MB 510231 0.5 mm, width ¼ 0.5 mm, n ¼ 72). This is much smaller than Etym.: From the host name, Dracaena, and -philum, Greek conidia on the specimens from China described above. for loving. Acervuli nigri, globosi vel irregulatim elongati. Setae sparsae in Based on a morphological examination of the type speci- hymenio, nigrae, 143–187 5–6.5 mm, laeves, 3–6 septatae. Cellu- men, Arx (1957) considered C. dracaenae to be a synonym lae conidiogenae hyalinae, heteroblasticae,18–32 4–8 mm. Coni- of C. gloeosporioides and our observations agree with that dia (20–)22–34(–38) (5.5–)6.5–9.5 (–10) mm (mean ¼ 29 8.5, conclusion. n ¼ 190, sd longitudo ¼ 3.2, latitudo ¼ 0.96), late clavata v. cylin- Specimen examined: Germany: Munich, in the greenhouse of the drica, hyalina. botanical garden, on faded leaves of Dracaena latifolia, Apr. 1895, Typus: China: on stems of Dracaena sp., APHIS interception San J.E. Weiss (M-0090064). Francisco, California 052750, 25 Sep. 2002, det. M.D. Petrillo, isol. d M. E. Palm MEP 1532 (holotypus BPI 871498, ex type culture CBS Colletotrichum dracaenae Petch, Ann. Roy. Bot. Gard. Perad. 9: 118199). 325 (1925), non Allescher 1902 nec Trinchieri 1909.

Acervuli on dying stems, numerous in discoloured areas, de- The fruiting bodies on the type specimen of this later hom- veloping subepidermally, surface exposed through irregularly onym have a very thin subhymenial layer that is only one or splitting of epidermal tissue that folds back, acervuli black, two layers thick. This is unlike those of Colletotrichum spp. globose to irregularly elongate, often coalescing, 210–630 mm Specimen examined: Sri Lanka: Peradeniya, on leaves of Dracaena in longest dimension. Cells at acervular surface mainly sanderiana, May 1904, T. Petch 6775 (K(M) 125641 d holotype of 5–6 mm diam forming textura prismatica, radially oriented to- C. dracaenae Petch, hom. illeg.). ward outer edge. Setae sparse, scattered in hymenium, black, 143–187 5–6.5 mm, straight to slightly irregularly rounded, Colletotrichum dracaenicola Sacc. & Trott, Syll. Fung. 22: 1205 becoming narrower and hyaline at rounded apex, smooth, (1913). 1402 D. F. Farr et al.

Fig 3 – Colletotrichum dracaenophilum. A. Conidiomata on natural substrata. BPI 842097. B. Section of conidiomata with setae produced on alfalfa stems in culture. CBS 118200. C. Section of conidiomata with conidiophores, conidia and setae produced on alfalfa stems in culture. CBS 118199 ex holotype culture. D. As above but without setae. CBS 119360. E. Conidia produced on alfalfa stems in culture. CBS 118200. F. Conidia on natural substrata. BPI 871498 Holotype. G–I. Glomerella sp., possible teleomorph of C. dracaenophilum. BPI 842097. G. Ascomata, longitudinal section. H–I. Asci and paraphyses. Bars: A–D, G–H [ 100 mm: E–F, I [ 10 mm. Colletotrichum on Agavaceae 1403

Synonym: Colletotrichum dracaenae Trinchieri. Rend. Accad. (mean ¼ 24.5 5.5 mm, sd length ¼ 2.5 mm, width ¼ 0.5 mm, Napoli. fasc. 8–12: 7 (1909), non Allescher 1902. n ¼ 69). Appressoria terminal or lateral, at first obpyriform, becoming irregularly lobed, 9–12 6–11 mm. The type specimen of C. dracaenicola could not be located, al- Ascomata on upper and lower surface of leaves in large, el- though requested from NAP and PORUN, nor is it housed at liptical, discoloured areas similar to those bearing acervuli, PAD (Gola 1930). Based on the description in Saccardo, this with or without a narrow, black margin, subepidermal, some- species has conidia 12–19 2–7 mm that are much smaller times partially erumpent, solitary, scattered to crowded or than those of C. dracaenophilum. aggregated, black, shiny when exposed, globose to ellipsoid, flattened. Ascomatal walls of thin-walled, brown cells, Colletotrichum dracaenae-fragrantis (H. Mori) Petr. & Syd., 9–15 mm diam. Paraphyses sparse, inflated, hyaline. Asci uni- Ann. Mycol. 23: 251 (1925). tunicate, narrowly clavate with a rounded apex and short Synonym: Macrophoma dracaenae-fragrantis H. Mori, Bull. Soc. stipe, 56–70 15–20 mm, with an indistinct apical ring in im- Bot. Ital. 3: 131 (1893). mature asci, 8-spored, obliquely seriate. Ascospores hyaline, non-septate, ellipsoidal, 15–22 4.5–6 mm. The type specimen for this name could not be located, how- Colonies 2.5–3.5 cm diam, slightly to thickly fluffy, vina- ever, based on the description in Saccardo, this species has ceous buff to hazel, slightly paler at margin, margin even, ol- conidia that are 5–12 2.5–3.5 mm, much smaller than those ivaceous grey toward a centre with slimy conidial masses, of C. dracaenophilum. An exsiccati specimen at BPI identified reverse smoke grey to grey olivaceous. No pigment in me- as this species was immature. dium. On CMD 2.3–2.8 cm diam, not fluffy, mouse grey toward Specimen examined: Romania:onDracaena fragrans Ker-Gawl, 15 centre, sporulating in small clumps; thick-walled cells devel- Aug. 1951, Savulescu, Myc. Roman. 1537 (BPI-bound exsiccati). oping in immersed mycelium. Host range: On living and recently killed leaves of Phormium Colletotrichum phormii (Henn.) D.F. Farr & Rossman, comb. cookeanum, Phormium sp. and P. tenax. nov. Geographic distribution: New Zealand, where the hosts are Fig 4A–F native, and elsewhere in temperate regions where Phormium, MycoBank no.: MB 499342 especially P. tenax, is cultivated, often in greenhouses, includ- Basionym: Fusarium phormii Henn, Verh. Bot. Ver. Prov. Brandenb ing Europe (Austria; Germany; Italy; Malta in Saccardo, 1915 as 40: 175 (1898). Gloeosporium phormii; Portugal in Almeida & Camara 1909 as Synonyms: Gloeosporium phormii (Henn.) Wollenw., Fus. Auto Cryptosporium rhodocyclum; Republic of Georgia in Dzhalago- Delin. No. 498 (1916), non Sacc. 1915. nia, 1965 as G. phormii; Spain in Unamuno, 1941 as Physalospora Synonym: Cryptosporium rhodocyclum Mont. ex Almeida & phormii; United Kingdom, specifically England and Scotland in Camara, Bol. Soc. Brot. 25: 190 (1909). Foister 1961 and Kirk & Spooner 1984), South Africa, and pos- Colletotrichum rhodocyclum (Mont.) Petr., Ann. Mycol. 25: 251 sibly in the USA (Florida based on Alfieri et al. 1984). (1927). Type specimens examined: Germany: Hortus Berolinensis: Kalthaus, Gloeosporidium rhodocyclum (Mont.) Ho¨ hn., Ann. Mycol. 18:92 on Phormium tenax, Apr. 1889 (B 70 0005220dholotype of Fusarium (1920). phormii; BPI-isotype); Breslau, Botan. Garten im Warmhause, on Gloeosporium phormii Sacc., Nuovo Giorn. Bot. Ital. n.s. 22:67 decaying leaf tips of P. tenax, on or before Mar. 1894 (Pilze Schle- (1915). siens: Type of Physalospora phormii not located). Additional specimens examined: Austria: Vorarlberg: Stella matu- Teleomorph: Glomerella phormii (J. Schro¨ t.) D.F. Farr & Ross- tina, Feldkirch, on P. tenax, 1902, S.J. Rick (S F39251, F39252, man, comb. nov. F39253 as Physalospora phormii, also issued as Rehm: Ascomyce- MycoBank no.: 510233 tun 1295b); Tirol: St Oswald, on leaves of P. tenax, 11 Jan. 1916, Basionym: Physalospora phormii J. Schro¨ t, in Cohn, Krypt.-Fl W. Pfaff, det. F. Buba´k (BPI 599638). d Germany: Berlin, Horticul- Schlesien 3: 347 (1894). tural Garden, on P. tenax, 3 May 1899, P. Hennings (S F39248 as Synonym: Hypostegium phormii (J. Schro¨ t.) Theiss., Verh. Zool.- Physalospora phormii); ‘Kgl. Botan. Garten’, on dying ends of bot. Ges. Wien 66: 384 (1916). leaves of P. tenax, coll. P. Hennings, Rehm, Ascomyceten 1295 Physalospora phormii (S P39246, S P39247); on Phormium sp., APHIS interception Port Orlando, Florida 007160, 6 Nov. 2000, W. Sheta, m On lower surfaces of living leaves, acervuli black, 280–420 m det. J. McKemy, isol. A. Rossman (AR 3546, CBS 118194, BPI 871494, long, no setae observed, scattered, subepidermal, developing Colletotrichum state only). d Italy: Tirolia: St Oswald, on P. tenax, in large, elliptical discolored areas, lesions (0.2–) 2.0– 8 Dec. 1915, W. Pfaff, det. F. Buba´k (BPI 599637 as Physalospora 3.7 0.5–0.9 cm, with generally well-defined, white to brown phormii); 11 Jan. 1916 (BPI 599638). d New Zealand: Wairoa: margin, broad to narrow pale green outer region, and brown Hawke’s Bay, on P. tenax, Apr. 1931, comm. P.G.M. Rhodes, det. to black in inner region, acervuli not splitting widely, produc- E.A. Hodgson (S F39240 as P. phormii); on Phormium, APHIS inter- ception Los Angeles, California 134866, 1 May 1997, M. A. Abdel- ing pale orange cirrhus of conidia. On upper surface acervuli shife, det. R. Schulteiss, isol. A. Rossman (AR 3389, CBS 118197, BPI less conspicuous, not in discoloured areas, developing below 746430); on Phormium, APHIS interception Los Angeles, California epidermis, erumpent through epidermis that remains at- 105828, 4 May 1993, N. Suzuki, isol. M.E. Palm (MEP 1334, CBS tached, surface sunken. Acervuli of a palisade of 2–5, dark cells 118201, BPI 802177); APHIS interception Los Angeles, California subtending conidiogenous cells. Conidiogenous cells cylindri- 093629, 1 May 1991, Y. Ishibashi (BPI 737566); APHIS interception cal, 13–26 4–5 mm. Conidia cylindrical to long fusiform, Los Angeles, California 099701, 11 May 1992, coll. J. Nelson, (BPI 802176); APHIS interception Los Angeles, California 105825, 28 gradually tapering to broadly truncate base, apex rounded, Apr. 1993, R. Heliczer (BPI 802175). d South Africa:onPhormium straight to slightly curved, (16.5–)19–29.5(–31) 4–7.5 mm cookeanum, APHIS interception San Francisco California 051697, 1404 D. F. Farr et al.

Fig 4 – Colletotrichum phormii. A. Conidiomata on lower surface of leaves. CBS 118194. B. Conidiomata produced in culture on alfalfa stems. CBS 118194. C. Conidiophores from natural substrata. B70005220. D. Appressoria produced in slide culture. CBS 118194. E. Conidia produced in culture on alfalfa stems. CBS 118194. F. Conidia on natural substrata. B700005220. G–I. Phaeosphaeriopsis phacidiomorpha Rabenhorst, Fungi Europaei 2337 as Sphaeria phacidiomorpha. G. Asci. H. Asci and pseu- doparaphyses. I. Ascospore released from . Bars: A–C [ 100 mm, D–I [ 10 mm.

13 Apr. 2000, L. Krekorian, det. M.E. Palm, isol. A. Rossman (AR a commercial crop in Devonshire, England in the spring of 3410, CBS 118195, BPI 871495); on Phormium, APHIS interception 1934 and later that year. Kinghorn (1936) published the only Miami, Florida 223143, 26 Feb. 2002, H. Ruiz, det. M.E. Palm, account of both the disease and the development of the fun- isol. A. Rossman (AR 3787, CBS 118191, BPI 871493). d United gus. Perithecia were reported to develop on leaves in 3–4 Kingdom; West Cornwall, St Ives, on P. tenax, Jul. 1931, det. W.B. Grove (S F39242 as Physalospora phormii); Scotland, on weeks; however, no perithecia developed in culture during P. tenax, W.O. Kinghorn (CBS 198.35 as Glomerella phacidiomorpha); this study. Of the three long-spored species of Colletotrichum same (CBS 199.35 as G. phacidiomorpha). specific to Agavaceae, C. phormii is the slowest growing in Notes: C. phormii is the best known of the long-spored spe- culture. cies of Colletotrichum on Agavaceae with most reports under In reviewing the nomenclature of this species it was dis- the name C. rhodocyclum following the thorough study of this covered that the description of the basionym Cryptosporium species by Kinghorn (1936). Since that report relatively little rhodocyclum, initially cited as ‘Montagne in litt.,’ was not val- has been published about this disease. C. phormii (as C. rhodo- idly published until Almeida & Camara did so in 1909, the cyclum or the teleomorph name G. phacidiomorpha) was date from with this epithet originates. Thus, the earliest reported to cause a disease on Phormium tenax cultivated as basionym for this species is Fusarium phormii Henn. 1899 Colletotrichum on Agavaceae 1405

Key to species of Colletotrichum on Agavaceae

1 Conidia generally less than 20 mm long...... 2 Conidia generally longer than 20 mm long...... 3

2(1) Conidia 12–17 3.5–5 mm, base truncate...... gloeosporioides Conidia 13–15.5 5–6 mm, base with a scar-like hilum...... boninense

3(1) Conidia averaging longer than 28 mm; colonies relatively fast-growing, 6–6.7 cm diam on PDA in 6 d. On Dracaena spp...... dracaenophilum Conidia averaging less than 28 mm long; colonies slow-growing, less than 5 cm diam on PDA in 6 d...... 4

4(3) Conidia 19–29.5 4–7 mm; setae generally lacking; colonies 3–3.2 cm on PDA in 6 d. On Phormium spp...... phormii Conidia 19–30.5 5.0–8.5 mm; setae abundant; colonies 3.5–4.5 cm on PDA in 6 d. On Agave and Furcraea spp...... agaves and that epithet is herein transferred to Colletotrichum. One Basionym: Glomerella phacidiomorpha (Ces.) Petr., Ann. Mycol. additional synonym, Gloeosporium phormii Sacc. 1915, is 25: 253 (1927). based on a specimen collected in Malta. Later Saccardo Synonyms: Sphaeria phacidiomorpha Ces., Fungi Europaei no. (1931) recognized the synonymy of his name with that of 2337 (1876). Fusarium phormii when he cited this name as G. phormii Didymella phacidiomorpha (Ces.) Sacc., Syll. Fung. 1: 559 (1882). (Henn.) Sacc. listing the specimen from Malta as well as Specimen examined: Italy: Horto Botanico Neapolitano, on leaves Henning’s type specimen from the Berlin Botanical Garden of Phormium tenax, Cesati, Rabenhorst, Fungi Europaei 2337 (BPI Sphaeria phacidiomorpha lectotypus hic in Germany. The later homonym of Gloeosporium phormii bound-lectotype of designatus). (Henn.) Wollenw. should perhaps be listed as G. phormii (Henn.) Buba´k. Wollenweber’s Fusarium delineatum autogra- The teleomorph of Colletotrichum phormii has often been re- phicum was published in 1916, however, it was not possible ferred to as G. phacidiomorpha following Kinghorn (1936). The to determine the precise date of publication. Buba´k’s combi- basionym Sphaeria phacidiomorpha was described in Rabenhor- nation was published on 10 October 1916 (Buba´k 1916), thus st’s Fungi Europaei (1876) and listed in Rabenhorst (1878). it is assumed that Wollenweber’s new combination is earlier However, an examination of the type specimen of Sphaeria ´ than that of Bubak. In addition, this authorship is accepted phacidiomorpha reveals that this taxon is not a Glomerella. Al- et al. by Kirk [ (http://www.indexfungoru- though appearing macroscopically similar to Glomerella, this Gloeosporium phormii m.org/Names/Names.asp) under ], al- fungus has bitunicate, obpyriform asci surrounded by abun- though von Arx (1957) attributed this new combination to dant, filiform, branched pseudoparaphyses with four- to ´ Bubak. The question of authorship in 1916 is moot because five-septate, hyaline ascospores that become distinctly con- this new combination was preceded by Saccardo’s publica- stricted at one of the middle septa and yellow at maturity tion of the same name in 1915. (Fig 4G–I). The conidia described as cylindric, hyaline, 3 mm The sexual state of Colletotrichum phormii was first de- long agree with the Microsphaeropsis-like anamorphs of Phaeos- scribed as Physalospora phormii, a name that is herein trans- phaeriopsis (Camara et al. 2003). Based on our examination of ferred to the genus Glomerella and serves as the name for the the type specimen, Sphaeria phacidiomorpha is placed in Phaeos- C. phormii teleomorph of . This name had previously been phaeriopsis in the , unrelated to Glomerella/ Hypostegium placed in the monotypic genus Theiss. 1916, Colletotrichum. thus Hypostegium is a later synonym of Glomerella Schrenk & H. Spauld. 1903. The teleomorph of C. phormii was regarded as Glomerella phacidiomorpha by Kinghorn (1936) but an examina- tion of the basionym of that name reveals that Sphaeria phacidio- Discussion morpha does not refer to a Glomerella (see below). Excluded names of Colletotrichum phormii and Glomerella Fungi in the genus Colletotrichum are present in soil and on phormii: plants as pathogens and endophytes throughout the world. Phyllosticta haematocycla was listed as a synonym of C. phor- Of the 40 recognized species, about 25 are represented by se- mii as C. rhodocyclum by Almeida and Camara (1909). However, quences in GenBank, although not all have ITS sequences. this name is now recognized as Phoma haematocycla (Berk.) Aa About 500 plant species are reported to be hosts of species of & Boerema (in de Gruyter et al. 1993) based on an examination Colletotrichum with most plants said to be infected with C. gloeo- of the type specimen of Phyllosticta haematocycla and is ac- sporioides. Recent publications have addressed the systematics cepted in Boerema et al. (2004). of species of Colletotrichum but often only taxa on one plant host are included, e.g. Fragaria (Martinez Culebras et al. 2003; Xiao Phaeosphaeriopsis phacidiomorpha (Ces.) D.F. Farr & M.E. et al. 2004), Rhododendron (Vinnere et al. 2002), or only one spe- Palm, comb. nov. cies of Colletotrichum is treated (Guerber et al. 2003; Horvath & Fig 4G–I Vargas, 2004). Many of the species defined by Sutton (1980) MycoBank no.: MB 510232 have yet to be sequenced and their concepts clarified. 1406 D. F. Farr et al.

Of the five species of Collectotrichum on Agavaceae, three Glomerella, unbranched true paraphyses develop from the peri- were restricted to specific genera of Agavaceae while the other thecial base among the ascogenous hyphae but dissolve as the two species have broad host ranges. Both Colletotrichum agaves asci develop and thus may not be evident in mature perithecia. and C. dracaenophilum appear to be host-specific taxa that are The asci are unitunicate, generally lacking an apical ring, al- distinct within the genus Colletotrichum. Neither of these though the apex of immature asci often appears thickened. As- taxa is closely related to any other species included in this mo- cospores are uniseriate, non-septate, although often guttulate, lecular analysis of Colletotrichum species. hyaline, generally ellipsoidal, and smooth-walled. Species of Based on the molecular sequence data presented here, Glomerella/Colletotrichum are placed in the . Phylo- C. phormii is most closely related to C. acutatum (Fig 1). Origi- genetic placement of this family has not been adequately de- nally described as a fruit-rotting fungus, C. acutatum is now termined although species of Glomerella/Colletotrichum seem known from a variety of plants (Farr et al. 2005; Peres et al. to be closely allied with Plectosphaerella/Plectosporium in the 2005). Many older reports of C. gloeosporioides as a plant path- (Huhndorf et al. 2004). Within Glomerella/Colleto- ogen may actually refer to C. acutatum. C. acutatum is one of trichum few morphological distinctions between taxa can be the most troublesome pathogens of along with made based on the sexual state alone. Morphological variabil- C. gloeosporioides (Xiao et al. 2004). Within what is increasingly ity among species of Glomerella/Colletotrichum is generally more regarded as the C. acutatum species complex (Guerber et al. evident in the asexual state as is the case with other ascomy- 2003; Peres et al. 2005; Talhinhas et al. 2002), one species has cetes, e.g., /Fusicoccum/Diplodia (Alves et al. been recognized as host-specific, namely C. lupini. C. phormii 2004); Calonectria/Cylindrocladium (Crous 2002); and Hypocrea/ also constitutes a distinct and host-specific species within Trichoderma (Chaverri & Samuels 2003). the C. acutatum lineage. Conidia of C. phormii (19–29 4– 7 mm) are morphologically distinct from those of C. acutatum (8–16 2.5–4 mm), which are considerably shorter and nar- Acknowledgements rower than those of C. phormii. In addition, colonies of C. acu- tatum on PDA often produce bright pink to reddish pigments, We would like to acknowledge the skilled technical assistance which are not produced by C. phormii. C. phormii is also mor- of Cindy Park. Malcolm DeCruise, Aimee Hyten and Franklin phologically distinct from C. lupini that produces conidia mea- Hendricks who were also extremely helpful in caring for the suring 12.5–15 4–45 mm(Nirenberg et al. 2002), considerably many isolates used in this study. We thank Jim Correll for smaller than those of C. phormii. cultures of Colletotrichum acutatum and the following reference C. boninense was described recently as a segregate of the collections: B, K, M, PC and S, from which specimens were C. gloeosporioides complex (Moriwaki et al. 2003) and is known graciously sent to us on loan. Finally, we appreciate the APHIS from a diversity of plant species including monocotyledonous, plant pathology identifiers, many listed as collectors in this dicotyledonous, herbaceous and woody plants. It is differenti- paper, for their diligence in safeguarding US agriculture and ated from C. gloeosporioides by conidial length-breadth ratio. for their professionalism and contribution to our knowledge C. boninense has relatively broad conidia 13–15.5 5–6 mmwith of the biodiversity of plant pathogenic fungi. They provided length:breadth 1.8–3.3, a hilum-like conidial base, and cream both the impetus and numerous specimens for this study. to orange-coloured colonies on PDA (Moriwaki et al. 2003). One ascospore isolate of Glomerella cingulata, the sexual state of C. gloeosporioides, from Dracaena fell within the C. gloeo- references sporioides group as expected, allied with two other sexual state isolates. The C. gloeosporioides-complex, including G. cingulata, includes strains from diverse hosts that function as both en- Alfieri jr SA, Langdon KR, Wehlburg C, Kimbrough JW, 1984. Index dophytes and as plant pathogens causing leaf spots. All of plant diseases in Florida (revised). Florida Department of Ag- riculture and Consumer Services, Division of Plant Industry Bulletin strains of C. gloeosporioides from Theobroma included in this 11: 1–389. study were isolated as endophytes as reported by Arnold Almeida JV, Sousa da Camara M, 1909. Contributiones ad myco- et al. (2003) while the strains from Pueraria (kudzu) are being floram Lusitaniae. Boletim de Sociedade Broteriana 24: 150–213. studied as potential biocontrol agents of this invasive weed Alves A, Correia A, Luque J, Phillips A, 2004. Botryosphaeria corticola (C. Boyette, pers. comm.). A thorough study of relationships sp. nov. on Quercus species, with notes and description of Bo- among fungi in the C. gloeosporioides-complex is needed. tryosphaeria stevensii and its anamorph, Diplodia mutila. Mycologia 96: 598–613. Anonymous, 1979. List of Plant Diseases in Taiwan. Plant Protection The sexual states of species of Colletotrichum are placed in Society, Republic of China. the holomorph genus Glomerella, although relatively few spe- Arnold AE, Mejı´a LC, Kyllo DA, Rojas EI, Maynard Z, Robbins N, cies are known to produce the sexual state. At present only Herre EA, 2003. Fungal endophytes limit pathogen damage in five species with a Glomerella state are recognized by Kirk a tropical tree. Proceedings of the National Academy of Sciences, et al. (2001), although 95 species names in Glomerella have USA 100: 15649–15654. been published. Glomerella produces solitary, uniloculate peri- von Arx JA, 1957. Die Arten der Gattung Colletotrichum Cda. Phy- thecia usually immersed in the substrata and becoming par- topathologische Zeitschrift 29: 413–468. Barthelet J, 1942. L’anthracnose des agaves. Annales des E´piphyties tially erumpent. The centrum development of G. cingulata 8: 111–118. was carefully described by Uecker (1994) who noted that this Boerema GH, de Gruyter J, Noordeloos ME, Hamers MEC, 2004. was similar to that of Plectosphaerella cucumerina (Uecker Phoma Identification Manual. Differentiation of Specific and Infra- 1993), the species most closely related to Glomerella.In specific Taxa in Culture. CABI Publishing, Wallingford. Colletotrichum on Agavaceae 1407

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