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Curvularia tsudae comb. nov. et nom. nov., formerly Pseudocochliobolus australiensis, and a revised synonymy for Curvularia australiensis
m y c o s c i e n c e x x x ( 2 0 1 4 ) 1 e5
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Curvularia tsudae comb. nov. et nom. nov., formerly Pseudocochliobolus australiensis, and a revised synonymy for Curvularia australiensis Hui Deng a, Yu Pei Tan b,c, Roger G. Shivas c, Yong Chun Niu a,* a
Key Laboratory of Microbial Resources, Ministry of Agriculture/Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, No. 12 South Zhongguancun Street, Haidian, Beijing, China b Microbiology, Department of Biology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands c Plant Pathology Herbarium, Biosecurity Queensland, Ecosciences Precinct, Dutton Park, Queensland 4102, Australia
article info
abstract
Article history:
Cultures originally identified as Drechslera australiensis, from seeds of Chloris gayana in
Received 5 September 2013
Japan, were the basis for Tsuda and Ueyama’s new combination, Bipolaris australiensis, and
Received in revised form
its associated sexual morph Pseudocochliobolus australiensis. By studying ex-type materials
7 February 2014
of both Drechslera australiensis, which was originally isolated from seeds of Oryza sativa in
Accepted 7 February 2014
Australia, and Pseudocochliobolus australiensis, we show by morphological and molecular
Available online xxx
phylogenetic analysis that these two specimens represent different species. Taxonomic
Keywords:
comb. nov. et nom. nov., together with a revised synonymy for Curvularia australiensis.
confusion is resolved by the transfer of Pseudocochliobolus australiensis to Curvularia tsudae Morphology
ª 2014 The Mycological Society of Japan. Published by Elsevier B.V. All rights reserved.
Nomenclature Phylogeny Pleosporales Taxonomy
1.
Introduction
Sivanesan (1987) provided the first comprehensive morphology-based taxonomic treatment for the species of Bipolaris Shoemaker (Shoemaker 1959), Curvularia Boedijn (Boedijn 1933) and Cochliobolus Drechsler (Drechsler 1934) that infect or were associated with grasses (Poaceae). Molecular phylogenetic analyses have since shown that these genera divide into two groups (Berbee et al. 1999; Manamgoda et al. 2012). Bipolaris and Cochliobolus species clustered in one of these groups along with their respective type species, (Bipolaris
maydis (Y. Nisik. & C. Miyake) Shoemaker and Cochliobolus heterostrophus (Drechsler) Drechsler), whereas Curvularia (including species named as Bipolaris, Cochliobolus, Pseudocochliobolus Tsuda, Ueyama & Nishih. and Curvularia) clustered in the other group, with its generic type, Curvularia lunata (Wakker) Boedijn (Manamgoda et al. 2012). Rossman et al. (2013) proposed the retention of Bipolaris over Cochliobolus as Bipolaris was more widely used and established in the plant pathology literature. Tsuda and Ueyama (1981) studied a fungus that had been isolated from the seeds of Chloris gayana Kunth provided by the Kyushu National Agriculture Research Station, Japan.
* Corresponding author. Tel./fax: þ86 010 82108647. E-mail address: [email protected] (Y.C. Niu). http://dx.doi.org/10.1016/j.myc.2014.02.002 1340-3540/ª 2014 The Mycological Society of Japan. Published by Elsevier B.V. All rights reserved.
Please cite this article in press as: Deng H, et al., Curvularia tsudae comb. nov. et nom. nov., formerly Pseudocochliobolus australiensis, and a revised synonymy for Curvularia australiensis, Mycoscience (2014), http://dx.doi.org/10.1016/j.myc.2014.02.002
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They believed that the fungus was Drechslera australiensis Bugnic. ex M.B. Ellis, which they subsequently transferred to Bipolaris australiensis (Bugnic. ex M.B. Ellis) Tsuda & Ueyama based on conidia morphology and bipolar conidial germination. Further, Tsuda and Ueyama (1981) established a new name, Pseudocochliobolus australiensis Tsuda & Ueyama, to represent the sexual morph of Bipolaris australiensis, which was produced in the mating experiments. Tsuda and Ueyama (1981) did not examine type specimens of Drechslera australiensis, but assumed that the isolates they obtained from seeds of Chloris gayana were authentic for the species. Subsequently Alcorn (1983) transferred Pseudocochliobolus australiensis to Cochliobolus australiensis (Tsuda & Ueyama) Alcorn, and Manamgoda et al. (2012) transferred Drechslera australiensis to Curvularia australiensis (M.B. Ellis) Manamgoda, L. Cai & K.D. Hyde. This has resulted in a synonymy that has placed Drechslera australiensis in five different genera (Manamgoda et al. 2012). In this study, we provide evidence based on molecular phylogeny and morphology that the fungus Tsuda and Ueyama (1981) studied was not authentic for Drechslera australiensis, but represented a novel species.
2.
Materials and methods
2.1.
Growth of cultures
The cultures studied were the ex-type of Drechslera australiensis (IMI 53994), the ex-paratype of Pseudocochliobolus australiensis (ATCC 44764), and two other strains of Cochliobolus
australiensis (BRIP 10967 and BRIP 10970) (Table 1). For morphological examination, cultures were grown on tap water agar with sterile pieces of wheat straw (TWA þ W). Conidia from cultures were mounted in lactic acid-glycerin aqueous solution (25% v/v lactic acid, 50% v/v glycerin) on glass slides for microscopic examination. For DNA extraction, these four cultures were grown on potato dextrose agar (PDA, Difco, Sparks, MD, USA) for 5e10 d.
2.2.
Molecular analyses
Total genomic DNA was extracted from fresh mycelium according to the methods given by Lee and Taylor (1990). The primers ITS4 and ITS5 (White et al. 1990) were used to amplify the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA (nrDNA). Partial region of the glyceraldehyde-3-phosphate dehydrogenase (GPDH) gene, large subunit (LSU) of the nrDNA, and translation elongation factor 1-a (EF-1a) gene were amplified using the primer pairs gpd1/ gpd2 (Berbee et al. 1999), EF1983/2218R and LR5/LROR, respectively (Schoch et al. 2009). The thermal cycling program was as follows: 95 C for 2 min, followed by 30 cycles of denaturation at 95 C for 1 min, annealing at 53 C for 1 min and elongation at 72 C for 2 min, with a final extension step of 72 C for 10 min. The ITS, LSU, GPDH, EF-1a DNA sequences were generated by Invitrogen (Beijing, China) and Beijing Sunbiotech (Beijing, China). The sequences were aligned using with Clustal X 1.81 (Thompson et al. 1997) and then further manually adjusted using BioEdit 5.0.6 (Hall 1999). Phylogenetic analyses were
Table 1 e Species, isolates and GenBank accession numbers of Curvularia and Bipolaris strains used in the phylogenetic analysis (ex-type cultures are in bold). Species
Locality
Strains no.a
GenBank accession no.
References
ITS
GDPH
LSU
EF-1
Curvularia coicis
Australia Australia Thailand Thailand Australia Vietnam Japan
BRIP 12790 CBS 280.91 MFLUCC 10-0703 MFLUCC 10-0711 IMI 53994 CBS 172.57 CBS 192.29
JN601034 JN601032 JX256420 JX256424 KC424595 JN601026 AF081447
JN600977 JN600974 JX276433 JX276436 KC747744 JN601036 AF081410
JN601000 JN600996 JX256387 JX256391 KC445296 JN600981 JN600984
JN601022 JN601018 JX266589 JX266593 KC503939 JN601003 JN601006
Curvularia ellisii Curvularia graminicola Curvularia hawaiiensis Curvularia heteropogonis Curvularia lunata Curvularia ovariicola Curvularia ravenelii Curvularia tripogonis Curvularia tsudae
Pakistan Australia Hawaii Australia USA Australia Australia Australia Japan
JN192375 JN192376 JN601029 JN192379 JX256429 JN192384 JN192386 JN192388 KC424596
JN600963 JN600964 JN600966 JN600969 JX276441 JN600976 JN600978 JN600980 KC747745
JN600985 JN600986 JN600988 JN600990 JX256396 JN600998 JN601001 JN601002 KC445297
JN601007 JN601008 JN601010 JN601013 JX266596 JN601020 JN601024 JN601025 KC503940
Curvularia tuberculata
Australia Australia India
CBS 193.62 BRIP 23186 CBS 173.57 CBS 284.91 CBS 730.96 CBS 470.90 BRIP 13165 BRIP 12375 ATCC 44764 ([Cg-157b) BRIP 10967 BRIP 10970 CBS 146.63
Manamgoda et al. 2011 Manamgoda et al. 2012 Manamgoda et al. 2012 Manamgoda et al. 2012 This study Manamgoda et al. 2012 Berbee et al. 1999; Manamgoda et al. 2011 Manamgoda et al. 2011 Manamgoda et al. 2012 Manamgoda et al. 2012 Manamgoda et al. 2011 Manamgoda et al. 2012 Manamgoda et al. 2011 Manamgoda et al. 2011 Manamgoda et al. 2011 This study
KC424604 KC424605 JX256433
KC747754 KC747755 JX276445
KC445306 KC445307 JX256401
KC503949 KC503950 JX266599
This study This study Manamgoda et al. 2012
Bipolaris peregianensis Bipolaris microlaenae Curvularia alcornii Curvularia asianensis Curvularia australiensis
a
ATCC: American Type Culture Collection, Manassas, Virginia. BRIP: Plant Pathology Herbarium, Brisbane, Queensland, Australia. CBS: Centraalbureau voor Schimmelcultures Fungal Biodiversity Centre, Utrecht, the Netherlands. IMI: CABI Bioscience Genetic Resource Collection, CABI Bioscience UK Centre, Egham, UK. Cg: M. Tsuda and A. Ueyama collectors’ no. MFLUCC: Mae Fae Luang University Culture Collection, Chiang Rai, Thailand. b Paratype of Pseudocochliobolus australiensis.
Please cite this article in press as: Deng H, et al., Curvularia tsudae comb. nov. et nom. nov., formerly Pseudocochliobolus australiensis, and a revised synonymy for Curvularia australiensis, Mycoscience (2014), http://dx.doi.org/10.1016/j.myc.2014.02.002
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performed in PAUP* v4.0b10 (Swofford 2002). Ambiguously aligned regions were excluded from all analyses. All gaps were treated as missing data and trees were viewed by Tree View (Page 1996). The concatenated alignment was deposited in TreeBASE (http://www.treebase.org/) (Study S14597), and the new taxon was deposited in MycoBank (http://www. mycobank.org/).
3.
Results and discussion
3.1.
Molecular phylogeny
The combined (ITS, GPDH, LSU and EF-1a) alignment was composed for 19 isolates (including the outgroups) (Table 1). The evolutionary relationship between isolates was analyzed using maximum parsimony. The inferred phylogenetic tree showed that the ex-type culture of Drechslera australiensis (IMI 53994) and the ex-paratype culture of Pseudocochliobolus australiensis (ATCC 44764) were not conspecific. Cochliobolus australiensis (BRIP 10967 and BRIP 10970) was conspecific with Pseudocochliobolus australiensis (Fig. 1). The phylogenetic tree also showed that these isolates all belong to the genus Curvularia (Fig. 1). As Drechslera australiensis was transferred to Curvularia australiensis by Manamgoda et al. (2012), a new name in Curvularia needs to be introduced for the proposed transfer of Pseudocochliobolus australiensis to avoid the making a homonym in Curvularia.
3.2.
3
Taxonomy
Curvularia australiensis (Bugnic. ex M.B. Ellis) Manamgoda, L. Cai & K.D. Hyde, Fungal Diversity 56: 140. 2012. hHelminthosporium australiense Bugnic., Rev. ge´n. Bot. 62: 242. 1956. (nom. inval. as nomen nudum, Art. 39.1 Melbourne Code). hDrechslera australiensis Bugnic. ex Subram. & B.L. Jain [as ‘australiense’], Curr. Sci. 35: 354. 1966. (nom. inval. as nomen nudum and a type specimen was not indicated, Arts 39.1 and 40.1 Melbourne Code). hDrechslera australiensis Bugnic. ex M.B. Ellis, Dematiaceous Hyphomycetes: 413. 1971. hBipolaris australiensis (Bugnic. ex M.B. Ellis) Tsuda & Ueyama, Mycologia 73: 90. 1981. On PDA colonies effuse, blackish brown with light gray aerial hyphae, velvety. On TWA þ W colonies with sparse aerial hyphae. Hyphae pale to mid brown, smooth, septate. Conidiophores single, flexuous, geniculate, septate, smooth, cylindrical, reddish brown, up to 235 6e15 mm. Conidiogenous cells with verruculose nodes. Conidia straight, ellipsoidal or oblong, rounded at the ends, reddish brown to black brown, usually 3- or rarely 4e5-distoseptate, 27e43 7.5e10 mm, 32 9 mm on average. Type: AUSTRALIA, New South Wales, Yanco, from seeds of Oryza sativa, Jan. 1951, F. Bugnicourt, IMI 53994 (lectotype). Isolectotypes: BRIP 12044, CBS 126973. Notes: Ellis (1971) validated Drechslera australiensis by providing a Latin description, and indicating a lectotype from
Fig. 1 e Phylogram generated from the maximum parsimony analysis based on combined genes of ITS, LSU, GPDH and EF1a sequence data. The tree is rooted to Bipolaris peregianensis Alcorn and B. microlaenae Alcorn. Bootstrap values of more than 50 are shown. The scale bar denotes 10 nucleotide substitutions. Please cite this article in press as: Deng H, et al., Curvularia tsudae comb. nov. et nom. nov., formerly Pseudocochliobolus australiensis, and a revised synonymy for Curvularia australiensis, Mycoscience (2014), http://dx.doi.org/10.1016/j.myc.2014.02.002
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one of Bugnicourt’s syntypes. Our morphological description of Curvularia australiensis is based on an isolate (CBS 172.57) from Vietnam, collected on seeds of Oryza sativa, in Dec. 1957 by F. Bugnicourt. This specimen was genetically identical to the ex-lectotype culture (IMI 53994) (Fig. 1, Table 1), which was sterile under our culture conditions. Tsuda and Ueyama (1981) provided a new name, Pseudocochliobolus australiensis, for the sexual morph of what they mistakenly thought was Drechslera australiensis. Transfer of P. australiensis to Curvularia is necessary to reflect the result of our phylogenetic analysis of an ex-paratype culture. A new species name is chosen as the epithet australiensis is not available as it would create a homonym. Curvularia tsudae (Tsuda & Ueyama) H. Deng, Y.P. Tan & R. G. Shivas, comb. nov. et nom. nov. Fig. 2. MycoBank no.: MB 805173. Replaced synonym: Pseudocochliobolus australiensis Tsuda & Ueyama, Mycologia 73: 92. 1981. hCochliobolus australiensis (Tsuda & Ueyama) Alcorn, Mycotaxon 16: 373. 1983. Non Curvularia australiensis (M.B. Ellis) Manamgoda, L. Cai & K.D. Hyde, Fungal Diversity 56: 140. 2012. Etymology: Named after the Japanese plant pathologist and mycologist Mitsuya Tsuda, who first described and named this fungus. On PDA colonies effuse, blackish with white aerial hyphae at the center, velvety. On TWA þ W colonies powdery gray aerial hyphae. Hyphae pale to mid brown, smooth, septate. Conidiophores single, flexuous, geniculate, septate, smooth, cylindrical, reddish brown, up to 165 mm 3e7 mm. Conidiogenous cells with verruculose nodes. Conidia straight, ellipsoidal or
fusoid, rounded at the ends, pale brown to mid brown, 3distoseptate, 17e30 7.5e10 mm, 26 9 mm on average (Fig. 2). Holotype: JAPAN, from seeds of Chloris gayana, H. Nakagawa, PRI-79001 (TNS-F-195405). Isotypes: BPI, CUP, IMI, L. Paratypes: PRI-79014 ¼ Cg-150-1 (TNS-F-195410), PRI-79015 ¼ Cg-157 (TNS-F-195411) ¼ ATCC 44764. Gene sequences ex-paratype strain ATCC 44764: KC424596 (ITS), KC747745 (GPDH), KC445297 (LSU), KC503940 (EF-1a). Additional specimens examined: AUSTRALIA, Queensland, Booie, from leaf tip blight of Chloris gayana, 12 Dec. 1972, J.L. Alcorn, BRIP 10967; Kenmore, from leaf lesion on Chloris gayana, 6 Jan. 1974, J.L. Alcorn, BRIP 10970. Notes: Tsuda and Ueyama (1981) established Pseudocochliobolus australiensis with holotype PRI-79001 (TNS-F-195405), which was deposited in the National Science Museum, Tokyo. We were unable to obtain the ex-holotype culture of Pseudocochliobolus australiensis but were able to locate an ex-paratype culture PRI-79015 (Cg-157, TNS-F-195411) held in the American Type Culture Collection (ATCC 44764), which Tsuda and Ueyama (1981) used in the mating experiments to demonstrate pseudothecium production and heterothallism, thereby verifying its authenticity. Recent synonymies for Curvularia australiensis (Manamgoda et al. 2012) and Cochliobolus australiensis (Manamgoda et al. 2011) are revised in light of our evidence that Tsuda and Ueyama (1981) did not examine an authentic isolate of Drechslera australiensis when establishing Pseudocochliobolus australiensis for its sexual morph. We have shown by molecular phylogenetic analysis that P. australiensis represents a species of Curvularia distinct from Curvularia australiensis. As the epithet ‘australiensis’ is no longer available, as this would create a homonym with Curvularia australiensis
Fig. 2 e Curvularia tsudae (ex-paratype ATCC 44764). A: Colony on PDA after 10 d at 23 C. B: Colony on TWA D W after 12 d at 23 C. C, D: Condiophore and conidia on TWA D W. Bars: 10 mm. Please cite this article in press as: Deng H, et al., Curvularia tsudae comb. nov. et nom. nov., formerly Pseudocochliobolus australiensis, and a revised synonymy for Curvularia australiensis, Mycoscience (2014), http://dx.doi.org/10.1016/j.myc.2014.02.002
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(M.B. Ellis) Manamgoda, L. Cai & K.D. Hyde, we have chosen the new name Curvularia tsudae. Curvularia tsudae differs morphologically from Curvularia australienis. The conidia of Curvularia tsudae are always 3distoseptate and average 26 mm in length, whereas Curvularia australienis has 3- or rarely 4e5-distoseptate conidia that average 32 mm in length. Furthermore, Curvularia tsudae has paler conidia than Curvularia australiensis. The literature shows that fungi attributed to Curvularia australiensis may be serious human pathogens (McGinnis et al. 1986; Flanagan and Bryceson 1997; Newell et al. 2006). In addition, there are several references in the literature to Curvularia australiensis as a pathogen on grasses (Poaceae), e.g., leaf spot on Pennisetum typhoides Stapf & C.E. Hubb (Chand and Singh 1966), and leaf blight on Cymbopogon winterianus Jowitt (Ramaiah and Chandrashekar 1981), as well as other plant hosts, e.g., pod rot of Senna angustifolia Batka (Saroj et al. 2011). Future molecular phylogenetic analyses may reveal other cryptic species morphologically similar to Curvularia australiensis and Curvularia tsudae. The International Code of Nomenclature for algae, fungi and plants (Melbourne Code) (McNeill et al. 2012) gives equal status to all legitimate names of asexual and sexual morphs for the purposes of priority. There is a potential for taxonomic confusion if the names of the asexual and sexual morphs are not based on the same type specimen as we have shown for Drechslera australiensis and Pseudocochliobolus australiensis. All anamorphic/teleomorphic fungi combinations warrant reexamination if based on different types.
Disclosures The authors declare no conflict of interest. All the experiments undertaken in this study comply with the current laws of the country in which they were performed.
Acknowledgments This study was supported by the National Natural Science Foundation of China (No. 30600010) and the Central Publicinterest Scientific Institution Basal Research Fund (No. 20104, 2012-9, 2013-28).
references
Alcorn JL, 1983. On the genera Cochliobolus and Pseudocochliobolus. Mycotaxon 16: 353e379. Berbee M, Pirseyedi M, Hubbard S, 1999. Cochliobolus phylogenetics and the origin of known, highly virulent pathogens, inferred from ITS and glyceraldehyde-3-phosphate dehydrogenase gene sequences. Mycologia 91: 964e977. Boedijn KB, 1933. Ueber Einige Phragmosporen Dematiazeen. Bulletin du Jardin botanique de Buitenzorg 13: 120e134. Chand JN, Singh B, 1966. A new Helminthosporium disease of Bajra. Current Science 35: 240. Drechsler C, 1934. Phytopathological and taxonomical aspects of Ophilobolus, Pyrenophora, Helminthosporium and a new genus Cochliobolus. Phytopathology 24: 953e983.
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Ellis MB, 1971. Dematiaceous hyphomycetes. Commonwealth Agricultural Bureaux, Kew. Flanagan KL, Bryceson AD, 1997. Disseminated infection due to Bipolaris australiensis in a young immunocompetent man: case report and review. Clinical Infectious Diseases 25: 311e313. Hall TA, 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Ser 41: 95e98. Lee SB, Taylor JW, 1990. Isolation of DNA from fungal mycelia and single spores. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds), PCR protocols: a guide to methods and applications. Academic Press, New York, pp 282e287. Manamgoda DS, Cai L, Bahkali AH, Chukeatirote E, Hyde KD, 2011. Cochliobolus: an overview and current status of species. Fungal Diversity 51: 3e42. Manamgoda DS, Cai L, McKenzie EHC, Crous PW, Madrid H, Chukeatirote E, Shivas RG, Tan YP, Hyde KD, 2012. A phylogenetic and taxonomic re-evaluation of the Bipolaris e Cochliobolus e Curvularia complex. Fungal Diversity 56: 131e144. McNeill J, Barrie FR, Buck WR, Demoulin V, Greuter W, Hawksworth DL, Herendeen PS, Knapp S, Marhold K, Prado J, Prud’homme van Reine WF, Smith GF, Wiersema JH, Turland N (eds), 2012. International Code of Nomenclature for algae, fungi, and plants (Melbourne Code), adopted by the Eighteenth International Botanical Congress Melbourne, Australia, July 2011. Koeltz Scientific Books, Ko¨nigstein. McGinnis MR, Rinaldi MG, Winn RE, 1986. Emerging agents of phaeohyphomycosis: pathogenic species of Bipolaris and Exserohilum. Journal of Clinical Microbiology 24: 250e259. Newell CK, Steinmetz RL, Brooks HL, 2006. Chronic postoperative endophthalmitis caused by Bipolaris australiensis. Retina 26: 109e110. Page RDM, 1996. Treeview. An application to display phylogenetic trees on personal computer. Computer Application in the Biosciences 12: 357e358. Ramaiah KS, Chandrashekar M, 1981. A new leaf blight disease of citronella grass. Current Science 50: 724e725. Rossman AY, Manamdoga DS, Hyde KD, 2013. Proposal to conserve the name Bipolaris against Cochliobolus (Ascomycota: Pleosporales: Pleosporaceae). Taxon 62: 1331e1332. Saroj A, Alam M, Qamar N, Khaliq A, Sattar A, 2011. First report of Bipolaris australiensis causing pod rot of senna in India. New Disease Reports 23: 28. Schoch C, Crous PW, Groenewald J, Boehm E, Burgess TI, DeGruyter J, De Hoog G, Dixon L, Grube M, Gueidan C, 2009. A class-wide phylogenetic assessment of Dothideomycetes. Studies in Mycology 64: 1e15. Shoemaker RA, 1959. Nomenclature of Drechslera and Bipolaris, grass parasites segregated from Helminthosporium. Canadian Journal of Botany 37: 879e887. Sivanesan A, 1987. Graminicolous species of Bipolaris, Curvularia, Drechslera, Exserohilum and their teleomorphs. Mycological Papers 158: 1e261. Swofford DL, 2002. PAUP*: phylogenetic analysis using parsimony (and other methods) 4.0 beta. Sinauer Associates, Sunderland, MA. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG, 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25: 4876e4882. Tsuda M, Ueyama A, 1981. Pseudocochliobolus australiensis, the ascigerous state of Bipolaris australiensis. Mycologia 73: 88e96. White TJ, Bruns T, Lee S, Taylor JW, 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds), PCR protocols: a guide to methods and applications. Academic Press, New York, pp 315e322.
Please cite this article in press as: Deng H, et al., Curvularia tsudae comb. nov. et nom. nov., formerly Pseudocochliobolus australiensis, and a revised synonymy for Curvularia australiensis, Mycoscience (2014), http://dx.doi.org/10.1016/j.myc.2014.02.002
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journal homepage: www.elsevier.com/locate/myc
Full paper
Curvularia tsudae comb. nov. et nom. nov., formerly Pseudocochliobolus australiensis, and a revised synonymy for Curvularia australiensis Hui Deng a, Yu Pei Tan b,c, Roger G. Shivas c, Yong Chun Niu a,* a
Key Laboratory of Microbial Resources, Ministry of Agriculture/Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, No. 12 South Zhongguancun Street, Haidian, Beijing, China b Microbiology, Department of Biology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands c Plant Pathology Herbarium, Biosecurity Queensland, Ecosciences Precinct, Dutton Park, Queensland 4102, Australia
article info
abstract
Article history:
Cultures originally identified as Drechslera australiensis, from seeds of Chloris gayana in
Received 5 September 2013
Japan, were the basis for Tsuda and Ueyama’s new combination, Bipolaris australiensis, and
Received in revised form
its associated sexual morph Pseudocochliobolus australiensis. By studying ex-type materials
7 February 2014
of both Drechslera australiensis, which was originally isolated from seeds of Oryza sativa in
Accepted 7 February 2014
Australia, and Pseudocochliobolus australiensis, we show by morphological and molecular
Available online xxx
phylogenetic analysis that these two specimens represent different species. Taxonomic
Keywords:
comb. nov. et nom. nov., together with a revised synonymy for Curvularia australiensis.
confusion is resolved by the transfer of Pseudocochliobolus australiensis to Curvularia tsudae Morphology
ª 2014 The Mycological Society of Japan. Published by Elsevier B.V. All rights reserved.
Nomenclature Phylogeny Pleosporales Taxonomy
1.
Introduction
Sivanesan (1987) provided the first comprehensive morphology-based taxonomic treatment for the species of Bipolaris Shoemaker (Shoemaker 1959), Curvularia Boedijn (Boedijn 1933) and Cochliobolus Drechsler (Drechsler 1934) that infect or were associated with grasses (Poaceae). Molecular phylogenetic analyses have since shown that these genera divide into two groups (Berbee et al. 1999; Manamgoda et al. 2012). Bipolaris and Cochliobolus species clustered in one of these groups along with their respective type species, (Bipolaris
maydis (Y. Nisik. & C. Miyake) Shoemaker and Cochliobolus heterostrophus (Drechsler) Drechsler), whereas Curvularia (including species named as Bipolaris, Cochliobolus, Pseudocochliobolus Tsuda, Ueyama & Nishih. and Curvularia) clustered in the other group, with its generic type, Curvularia lunata (Wakker) Boedijn (Manamgoda et al. 2012). Rossman et al. (2013) proposed the retention of Bipolaris over Cochliobolus as Bipolaris was more widely used and established in the plant pathology literature. Tsuda and Ueyama (1981) studied a fungus that had been isolated from the seeds of Chloris gayana Kunth provided by the Kyushu National Agriculture Research Station, Japan.
* Corresponding author. Tel./fax: þ86 010 82108647. E-mail address: [email protected] (Y.C. Niu). http://dx.doi.org/10.1016/j.myc.2014.02.002 1340-3540/ª 2014 The Mycological Society of Japan. Published by Elsevier B.V. All rights reserved.
Please cite this article in press as: Deng H, et al., Curvularia tsudae comb. nov. et nom. nov., formerly Pseudocochliobolus australiensis, and a revised synonymy for Curvularia australiensis, Mycoscience (2014), http://dx.doi.org/10.1016/j.myc.2014.02.002
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They believed that the fungus was Drechslera australiensis Bugnic. ex M.B. Ellis, which they subsequently transferred to Bipolaris australiensis (Bugnic. ex M.B. Ellis) Tsuda & Ueyama based on conidia morphology and bipolar conidial germination. Further, Tsuda and Ueyama (1981) established a new name, Pseudocochliobolus australiensis Tsuda & Ueyama, to represent the sexual morph of Bipolaris australiensis, which was produced in the mating experiments. Tsuda and Ueyama (1981) did not examine type specimens of Drechslera australiensis, but assumed that the isolates they obtained from seeds of Chloris gayana were authentic for the species. Subsequently Alcorn (1983) transferred Pseudocochliobolus australiensis to Cochliobolus australiensis (Tsuda & Ueyama) Alcorn, and Manamgoda et al. (2012) transferred Drechslera australiensis to Curvularia australiensis (M.B. Ellis) Manamgoda, L. Cai & K.D. Hyde. This has resulted in a synonymy that has placed Drechslera australiensis in five different genera (Manamgoda et al. 2012). In this study, we provide evidence based on molecular phylogeny and morphology that the fungus Tsuda and Ueyama (1981) studied was not authentic for Drechslera australiensis, but represented a novel species.
2.
Materials and methods
2.1.
Growth of cultures
The cultures studied were the ex-type of Drechslera australiensis (IMI 53994), the ex-paratype of Pseudocochliobolus australiensis (ATCC 44764), and two other strains of Cochliobolus
australiensis (BRIP 10967 and BRIP 10970) (Table 1). For morphological examination, cultures were grown on tap water agar with sterile pieces of wheat straw (TWA þ W). Conidia from cultures were mounted in lactic acid-glycerin aqueous solution (25% v/v lactic acid, 50% v/v glycerin) on glass slides for microscopic examination. For DNA extraction, these four cultures were grown on potato dextrose agar (PDA, Difco, Sparks, MD, USA) for 5e10 d.
2.2.
Molecular analyses
Total genomic DNA was extracted from fresh mycelium according to the methods given by Lee and Taylor (1990). The primers ITS4 and ITS5 (White et al. 1990) were used to amplify the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA (nrDNA). Partial region of the glyceraldehyde-3-phosphate dehydrogenase (GPDH) gene, large subunit (LSU) of the nrDNA, and translation elongation factor 1-a (EF-1a) gene were amplified using the primer pairs gpd1/ gpd2 (Berbee et al. 1999), EF1983/2218R and LR5/LROR, respectively (Schoch et al. 2009). The thermal cycling program was as follows: 95 C for 2 min, followed by 30 cycles of denaturation at 95 C for 1 min, annealing at 53 C for 1 min and elongation at 72 C for 2 min, with a final extension step of 72 C for 10 min. The ITS, LSU, GPDH, EF-1a DNA sequences were generated by Invitrogen (Beijing, China) and Beijing Sunbiotech (Beijing, China). The sequences were aligned using with Clustal X 1.81 (Thompson et al. 1997) and then further manually adjusted using BioEdit 5.0.6 (Hall 1999). Phylogenetic analyses were
Table 1 e Species, isolates and GenBank accession numbers of Curvularia and Bipolaris strains used in the phylogenetic analysis (ex-type cultures are in bold). Species
Locality
Strains no.a
GenBank accession no.
References
ITS
GDPH
LSU
EF-1
Curvularia coicis
Australia Australia Thailand Thailand Australia Vietnam Japan
BRIP 12790 CBS 280.91 MFLUCC 10-0703 MFLUCC 10-0711 IMI 53994 CBS 172.57 CBS 192.29
JN601034 JN601032 JX256420 JX256424 KC424595 JN601026 AF081447
JN600977 JN600974 JX276433 JX276436 KC747744 JN601036 AF081410
JN601000 JN600996 JX256387 JX256391 KC445296 JN600981 JN600984
JN601022 JN601018 JX266589 JX266593 KC503939 JN601003 JN601006
Curvularia ellisii Curvularia graminicola Curvularia hawaiiensis Curvularia heteropogonis Curvularia lunata Curvularia ovariicola Curvularia ravenelii Curvularia tripogonis Curvularia tsudae
Pakistan Australia Hawaii Australia USA Australia Australia Australia Japan
JN192375 JN192376 JN601029 JN192379 JX256429 JN192384 JN192386 JN192388 KC424596
JN600963 JN600964 JN600966 JN600969 JX276441 JN600976 JN600978 JN600980 KC747745
JN600985 JN600986 JN600988 JN600990 JX256396 JN600998 JN601001 JN601002 KC445297
JN601007 JN601008 JN601010 JN601013 JX266596 JN601020 JN601024 JN601025 KC503940
Curvularia tuberculata
Australia Australia India
CBS 193.62 BRIP 23186 CBS 173.57 CBS 284.91 CBS 730.96 CBS 470.90 BRIP 13165 BRIP 12375 ATCC 44764 ([Cg-157b) BRIP 10967 BRIP 10970 CBS 146.63
Manamgoda et al. 2011 Manamgoda et al. 2012 Manamgoda et al. 2012 Manamgoda et al. 2012 This study Manamgoda et al. 2012 Berbee et al. 1999; Manamgoda et al. 2011 Manamgoda et al. 2011 Manamgoda et al. 2012 Manamgoda et al. 2012 Manamgoda et al. 2011 Manamgoda et al. 2012 Manamgoda et al. 2011 Manamgoda et al. 2011 Manamgoda et al. 2011 This study
KC424604 KC424605 JX256433
KC747754 KC747755 JX276445
KC445306 KC445307 JX256401
KC503949 KC503950 JX266599
This study This study Manamgoda et al. 2012
Bipolaris peregianensis Bipolaris microlaenae Curvularia alcornii Curvularia asianensis Curvularia australiensis
a
ATCC: American Type Culture Collection, Manassas, Virginia. BRIP: Plant Pathology Herbarium, Brisbane, Queensland, Australia. CBS: Centraalbureau voor Schimmelcultures Fungal Biodiversity Centre, Utrecht, the Netherlands. IMI: CABI Bioscience Genetic Resource Collection, CABI Bioscience UK Centre, Egham, UK. Cg: M. Tsuda and A. Ueyama collectors’ no. MFLUCC: Mae Fae Luang University Culture Collection, Chiang Rai, Thailand. b Paratype of Pseudocochliobolus australiensis.
Please cite this article in press as: Deng H, et al., Curvularia tsudae comb. nov. et nom. nov., formerly Pseudocochliobolus australiensis, and a revised synonymy for Curvularia australiensis, Mycoscience (2014), http://dx.doi.org/10.1016/j.myc.2014.02.002
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performed in PAUP* v4.0b10 (Swofford 2002). Ambiguously aligned regions were excluded from all analyses. All gaps were treated as missing data and trees were viewed by Tree View (Page 1996). The concatenated alignment was deposited in TreeBASE (http://www.treebase.org/) (Study S14597), and the new taxon was deposited in MycoBank (http://www. mycobank.org/).
3.
Results and discussion
3.1.
Molecular phylogeny
The combined (ITS, GPDH, LSU and EF-1a) alignment was composed for 19 isolates (including the outgroups) (Table 1). The evolutionary relationship between isolates was analyzed using maximum parsimony. The inferred phylogenetic tree showed that the ex-type culture of Drechslera australiensis (IMI 53994) and the ex-paratype culture of Pseudocochliobolus australiensis (ATCC 44764) were not conspecific. Cochliobolus australiensis (BRIP 10967 and BRIP 10970) was conspecific with Pseudocochliobolus australiensis (Fig. 1). The phylogenetic tree also showed that these isolates all belong to the genus Curvularia (Fig. 1). As Drechslera australiensis was transferred to Curvularia australiensis by Manamgoda et al. (2012), a new name in Curvularia needs to be introduced for the proposed transfer of Pseudocochliobolus australiensis to avoid the making a homonym in Curvularia.
3.2.
3
Taxonomy
Curvularia australiensis (Bugnic. ex M.B. Ellis) Manamgoda, L. Cai & K.D. Hyde, Fungal Diversity 56: 140. 2012. hHelminthosporium australiense Bugnic., Rev. ge´n. Bot. 62: 242. 1956. (nom. inval. as nomen nudum, Art. 39.1 Melbourne Code). hDrechslera australiensis Bugnic. ex Subram. & B.L. Jain [as ‘australiense’], Curr. Sci. 35: 354. 1966. (nom. inval. as nomen nudum and a type specimen was not indicated, Arts 39.1 and 40.1 Melbourne Code). hDrechslera australiensis Bugnic. ex M.B. Ellis, Dematiaceous Hyphomycetes: 413. 1971. hBipolaris australiensis (Bugnic. ex M.B. Ellis) Tsuda & Ueyama, Mycologia 73: 90. 1981. On PDA colonies effuse, blackish brown with light gray aerial hyphae, velvety. On TWA þ W colonies with sparse aerial hyphae. Hyphae pale to mid brown, smooth, septate. Conidiophores single, flexuous, geniculate, septate, smooth, cylindrical, reddish brown, up to 235 6e15 mm. Conidiogenous cells with verruculose nodes. Conidia straight, ellipsoidal or oblong, rounded at the ends, reddish brown to black brown, usually 3- or rarely 4e5-distoseptate, 27e43 7.5e10 mm, 32 9 mm on average. Type: AUSTRALIA, New South Wales, Yanco, from seeds of Oryza sativa, Jan. 1951, F. Bugnicourt, IMI 53994 (lectotype). Isolectotypes: BRIP 12044, CBS 126973. Notes: Ellis (1971) validated Drechslera australiensis by providing a Latin description, and indicating a lectotype from
Fig. 1 e Phylogram generated from the maximum parsimony analysis based on combined genes of ITS, LSU, GPDH and EF1a sequence data. The tree is rooted to Bipolaris peregianensis Alcorn and B. microlaenae Alcorn. Bootstrap values of more than 50 are shown. The scale bar denotes 10 nucleotide substitutions. Please cite this article in press as: Deng H, et al., Curvularia tsudae comb. nov. et nom. nov., formerly Pseudocochliobolus australiensis, and a revised synonymy for Curvularia australiensis, Mycoscience (2014), http://dx.doi.org/10.1016/j.myc.2014.02.002
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one of Bugnicourt’s syntypes. Our morphological description of Curvularia australiensis is based on an isolate (CBS 172.57) from Vietnam, collected on seeds of Oryza sativa, in Dec. 1957 by F. Bugnicourt. This specimen was genetically identical to the ex-lectotype culture (IMI 53994) (Fig. 1, Table 1), which was sterile under our culture conditions. Tsuda and Ueyama (1981) provided a new name, Pseudocochliobolus australiensis, for the sexual morph of what they mistakenly thought was Drechslera australiensis. Transfer of P. australiensis to Curvularia is necessary to reflect the result of our phylogenetic analysis of an ex-paratype culture. A new species name is chosen as the epithet australiensis is not available as it would create a homonym. Curvularia tsudae (Tsuda & Ueyama) H. Deng, Y.P. Tan & R. G. Shivas, comb. nov. et nom. nov. Fig. 2. MycoBank no.: MB 805173. Replaced synonym: Pseudocochliobolus australiensis Tsuda & Ueyama, Mycologia 73: 92. 1981. hCochliobolus australiensis (Tsuda & Ueyama) Alcorn, Mycotaxon 16: 373. 1983. Non Curvularia australiensis (M.B. Ellis) Manamgoda, L. Cai & K.D. Hyde, Fungal Diversity 56: 140. 2012. Etymology: Named after the Japanese plant pathologist and mycologist Mitsuya Tsuda, who first described and named this fungus. On PDA colonies effuse, blackish with white aerial hyphae at the center, velvety. On TWA þ W colonies powdery gray aerial hyphae. Hyphae pale to mid brown, smooth, septate. Conidiophores single, flexuous, geniculate, septate, smooth, cylindrical, reddish brown, up to 165 mm 3e7 mm. Conidiogenous cells with verruculose nodes. Conidia straight, ellipsoidal or
fusoid, rounded at the ends, pale brown to mid brown, 3distoseptate, 17e30 7.5e10 mm, 26 9 mm on average (Fig. 2). Holotype: JAPAN, from seeds of Chloris gayana, H. Nakagawa, PRI-79001 (TNS-F-195405). Isotypes: BPI, CUP, IMI, L. Paratypes: PRI-79014 ¼ Cg-150-1 (TNS-F-195410), PRI-79015 ¼ Cg-157 (TNS-F-195411) ¼ ATCC 44764. Gene sequences ex-paratype strain ATCC 44764: KC424596 (ITS), KC747745 (GPDH), KC445297 (LSU), KC503940 (EF-1a). Additional specimens examined: AUSTRALIA, Queensland, Booie, from leaf tip blight of Chloris gayana, 12 Dec. 1972, J.L. Alcorn, BRIP 10967; Kenmore, from leaf lesion on Chloris gayana, 6 Jan. 1974, J.L. Alcorn, BRIP 10970. Notes: Tsuda and Ueyama (1981) established Pseudocochliobolus australiensis with holotype PRI-79001 (TNS-F-195405), which was deposited in the National Science Museum, Tokyo. We were unable to obtain the ex-holotype culture of Pseudocochliobolus australiensis but were able to locate an ex-paratype culture PRI-79015 (Cg-157, TNS-F-195411) held in the American Type Culture Collection (ATCC 44764), which Tsuda and Ueyama (1981) used in the mating experiments to demonstrate pseudothecium production and heterothallism, thereby verifying its authenticity. Recent synonymies for Curvularia australiensis (Manamgoda et al. 2012) and Cochliobolus australiensis (Manamgoda et al. 2011) are revised in light of our evidence that Tsuda and Ueyama (1981) did not examine an authentic isolate of Drechslera australiensis when establishing Pseudocochliobolus australiensis for its sexual morph. We have shown by molecular phylogenetic analysis that P. australiensis represents a species of Curvularia distinct from Curvularia australiensis. As the epithet ‘australiensis’ is no longer available, as this would create a homonym with Curvularia australiensis
Fig. 2 e Curvularia tsudae (ex-paratype ATCC 44764). A: Colony on PDA after 10 d at 23 C. B: Colony on TWA D W after 12 d at 23 C. C, D: Condiophore and conidia on TWA D W. Bars: 10 mm. Please cite this article in press as: Deng H, et al., Curvularia tsudae comb. nov. et nom. nov., formerly Pseudocochliobolus australiensis, and a revised synonymy for Curvularia australiensis, Mycoscience (2014), http://dx.doi.org/10.1016/j.myc.2014.02.002
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(M.B. Ellis) Manamgoda, L. Cai & K.D. Hyde, we have chosen the new name Curvularia tsudae. Curvularia tsudae differs morphologically from Curvularia australienis. The conidia of Curvularia tsudae are always 3distoseptate and average 26 mm in length, whereas Curvularia australienis has 3- or rarely 4e5-distoseptate conidia that average 32 mm in length. Furthermore, Curvularia tsudae has paler conidia than Curvularia australiensis. The literature shows that fungi attributed to Curvularia australiensis may be serious human pathogens (McGinnis et al. 1986; Flanagan and Bryceson 1997; Newell et al. 2006). In addition, there are several references in the literature to Curvularia australiensis as a pathogen on grasses (Poaceae), e.g., leaf spot on Pennisetum typhoides Stapf & C.E. Hubb (Chand and Singh 1966), and leaf blight on Cymbopogon winterianus Jowitt (Ramaiah and Chandrashekar 1981), as well as other plant hosts, e.g., pod rot of Senna angustifolia Batka (Saroj et al. 2011). Future molecular phylogenetic analyses may reveal other cryptic species morphologically similar to Curvularia australiensis and Curvularia tsudae. The International Code of Nomenclature for algae, fungi and plants (Melbourne Code) (McNeill et al. 2012) gives equal status to all legitimate names of asexual and sexual morphs for the purposes of priority. There is a potential for taxonomic confusion if the names of the asexual and sexual morphs are not based on the same type specimen as we have shown for Drechslera australiensis and Pseudocochliobolus australiensis. All anamorphic/teleomorphic fungi combinations warrant reexamination if based on different types.
Disclosures The authors declare no conflict of interest. All the experiments undertaken in this study comply with the current laws of the country in which they were performed.
Acknowledgments This study was supported by the National Natural Science Foundation of China (No. 30600010) and the Central Publicinterest Scientific Institution Basal Research Fund (No. 20104, 2012-9, 2013-28).
references
Alcorn JL, 1983. On the genera Cochliobolus and Pseudocochliobolus. Mycotaxon 16: 353e379. Berbee M, Pirseyedi M, Hubbard S, 1999. Cochliobolus phylogenetics and the origin of known, highly virulent pathogens, inferred from ITS and glyceraldehyde-3-phosphate dehydrogenase gene sequences. Mycologia 91: 964e977. Boedijn KB, 1933. Ueber Einige Phragmosporen Dematiazeen. Bulletin du Jardin botanique de Buitenzorg 13: 120e134. Chand JN, Singh B, 1966. A new Helminthosporium disease of Bajra. Current Science 35: 240. Drechsler C, 1934. Phytopathological and taxonomical aspects of Ophilobolus, Pyrenophora, Helminthosporium and a new genus Cochliobolus. Phytopathology 24: 953e983.
5
Ellis MB, 1971. Dematiaceous hyphomycetes. Commonwealth Agricultural Bureaux, Kew. Flanagan KL, Bryceson AD, 1997. Disseminated infection due to Bipolaris australiensis in a young immunocompetent man: case report and review. Clinical Infectious Diseases 25: 311e313. Hall TA, 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Ser 41: 95e98. Lee SB, Taylor JW, 1990. Isolation of DNA from fungal mycelia and single spores. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds), PCR protocols: a guide to methods and applications. Academic Press, New York, pp 282e287. Manamgoda DS, Cai L, Bahkali AH, Chukeatirote E, Hyde KD, 2011. Cochliobolus: an overview and current status of species. Fungal Diversity 51: 3e42. Manamgoda DS, Cai L, McKenzie EHC, Crous PW, Madrid H, Chukeatirote E, Shivas RG, Tan YP, Hyde KD, 2012. A phylogenetic and taxonomic re-evaluation of the Bipolaris e Cochliobolus e Curvularia complex. Fungal Diversity 56: 131e144. McNeill J, Barrie FR, Buck WR, Demoulin V, Greuter W, Hawksworth DL, Herendeen PS, Knapp S, Marhold K, Prado J, Prud’homme van Reine WF, Smith GF, Wiersema JH, Turland N (eds), 2012. International Code of Nomenclature for algae, fungi, and plants (Melbourne Code), adopted by the Eighteenth International Botanical Congress Melbourne, Australia, July 2011. Koeltz Scientific Books, Ko¨nigstein. McGinnis MR, Rinaldi MG, Winn RE, 1986. Emerging agents of phaeohyphomycosis: pathogenic species of Bipolaris and Exserohilum. Journal of Clinical Microbiology 24: 250e259. Newell CK, Steinmetz RL, Brooks HL, 2006. Chronic postoperative endophthalmitis caused by Bipolaris australiensis. Retina 26: 109e110. Page RDM, 1996. Treeview. An application to display phylogenetic trees on personal computer. Computer Application in the Biosciences 12: 357e358. Ramaiah KS, Chandrashekar M, 1981. A new leaf blight disease of citronella grass. Current Science 50: 724e725. Rossman AY, Manamdoga DS, Hyde KD, 2013. Proposal to conserve the name Bipolaris against Cochliobolus (Ascomycota: Pleosporales: Pleosporaceae). Taxon 62: 1331e1332. Saroj A, Alam M, Qamar N, Khaliq A, Sattar A, 2011. First report of Bipolaris australiensis causing pod rot of senna in India. New Disease Reports 23: 28. Schoch C, Crous PW, Groenewald J, Boehm E, Burgess TI, DeGruyter J, De Hoog G, Dixon L, Grube M, Gueidan C, 2009. A class-wide phylogenetic assessment of Dothideomycetes. Studies in Mycology 64: 1e15. Shoemaker RA, 1959. Nomenclature of Drechslera and Bipolaris, grass parasites segregated from Helminthosporium. Canadian Journal of Botany 37: 879e887. Sivanesan A, 1987. Graminicolous species of Bipolaris, Curvularia, Drechslera, Exserohilum and their teleomorphs. Mycological Papers 158: 1e261. Swofford DL, 2002. PAUP*: phylogenetic analysis using parsimony (and other methods) 4.0 beta. Sinauer Associates, Sunderland, MA. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG, 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25: 4876e4882. Tsuda M, Ueyama A, 1981. Pseudocochliobolus australiensis, the ascigerous state of Bipolaris australiensis. Mycologia 73: 88e96. White TJ, Bruns T, Lee S, Taylor JW, 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds), PCR protocols: a guide to methods and applications. Academic Press, New York, pp 315e322.
Please cite this article in press as: Deng H, et al., Curvularia tsudae comb. nov. et nom. nov., formerly Pseudocochliobolus australiensis, and a revised synonymy for Curvularia australiensis, Mycoscience (2014), http://dx.doi.org/10.1016/j.myc.2014.02.002