Sequence Data Reveals Phylogenetic Affinities of Fungal Anamorphs Bahusutrabeeja, Diplococcium, Natarajania, Paliphora, Polyschema, Rattania and Spadicoides
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Fungal Diversity (2010) 44:161–169 DOI 10.1007/s13225-010-0059-8 Sequence data reveals phylogenetic affinities of fungal anamorphs Bahusutrabeeja, Diplococcium, Natarajania, Paliphora, Polyschema, Rattania and Spadicoides Belle Damodara Shenoy & Rajesh Jeewon & Hongkai Wang & Kaur Amandeep & Wellcome H. Ho & Darbhe Jayarama Bhat & Pedro W. Crous & Kevin D. Hyde Received: 2 July 2010 /Accepted: 24 August 2010 /Published online: 11 September 2010 # Kevin D. Hyde 2010 Abstract Partial 28S rRNA gene sequence-data of the monophyletic lineage and are related to Lentithecium strains of the anamorphic genera Bahusutrabeeja, Diplo- fluviatile and Leptosphaeria calvescens in Pleosporales coccium, Natarajania, Paliphora, Polyschema, Rattania (Dothideomycetes). DNA sequence analysis also suggests and Spadicoides were analysed to predict their phylogenetic that Paliphora intermedia is a member of Chaetosphaer- relationships and taxonomic placement within the Ascomy- iaceae (Sordariomycetes). The type species of Bahusu- cota. Results indicate that Diplococcium and morphologi- trabeeja, B. dwaya, is phylogenetically related to cally similar genera, i.e. Spadicoides, Paliphora and Neodeightonia (=Botryosphaeria) subglobosa in Botryos- Polyschema do not share a recent common ancestor. The phaeriales (Dothideomycetes). Monotypic genera Natar- type species of Diplococcium, D. spicatum is referred to ajania and Rattania are phylogenetically related to Helotiales (Leotiomycetes). The placement of Spadicoides members of Diaporthales and Chaetosphaeriales, respec- bina, the type of the genus, is unresolved but it is shown to tively. Future studies with extended gene datasets and type be closely associated with Porosphaerella species, which strains are required to resolve many novel but morpholog- are sister taxa to Coniochaetales (Sordariomycetes). Three ically unexplainable phylogenetic scenarios revealed from Polyschema species analysed in this study represent a this study. It is increasingly becoming evident that a fungal B. D. Shenoy (*) : K. Amandeep D. J. Bhat Microbial Type Culture Collection and Gene Bank, Institute of Department of Botany, Goa University, Microbial Technology (CSIR), Taleigao Plateau, Goa 403 206, India Sector 39-A, Chandigarh 160 036, India e-mail: [email protected] P. W. Crous R. Jeewon (*) CBS-KNAW Fungal Biodiversity Centre, Department of Health Sciences, Faculty of Science, University of P.O. Box 85167, 3508 AD Utrecht, The Netherlands Mauritius, Reduit, Mauritius e-mail: [email protected] K. D. Hyde School of Science, Mae Fah Luang University, H. Wang Tasud, Biotechnology Institute, Zhejiang University, Chiang Rai, Thailand Hangzhou 310 029, People’s Republic of China W. H. Ho Plant Health and Environment Laboratory, Biosecurity New K. D. Hyde Zealand, Ministry of Agriculture and Forestry, Botany and Microbiology Department, College of Science, King 231, Morrin Road, St. Johns, P.O. Box 2095, Auckland 1140, Saud University, New Zealand Riyadh, Saudi Arabia 162 Fungal Diversity (2010) 44:161–169 lineage may include a mosaic of anamorphs, teleomorphs have thick, black or brown bands at the septa. The type and pleomorphs whose morphologies may not always be species of Diplococcium (i.e. D. spicatum Grove) and correlated. It is therefore suggested that where possible all Spadicoides (i.e. S. bina S. Hughes) differ mainly in new species descriptions, whether teleomorphic, anamor- branching of conidiophores and catenation of conidia. phic or pleomorphic, should include DNA sequence-data to Diplococcium spicatum produces catenate conidia on facilitate amalgamation of anamorphic and pleomorphic branched conidiophores, whereas Spadicoides bina produ- genera in a single phylogenetic classification system. ces solitary conidia on unbranched conidiophores (Hughes 1958; Ellis 1963; Sinclair et al. 1985; Goh and Hyde 1996). Keywords Ascomycetes . Asexual fungi . Hyphomycetes . The conidiophore branching was considered to be Molecular phylogeny. Taxonomy taxonomically more important at the generic level than conidial catenation by earlier mycologists (e.g. Ellis 1963, 1971a, b, 1972;Wang1976;WangandSutton Introduction 1982). Sinclair et al. (1985), however, stated that at the generic level, conidiophore branching is taxonomically The phenomenon of pleomorphism in fungi, i.e. occurrence less important than conidial catenation, and emended the of dual or multiple morphological forms of a fungal species generic description of Spadicoides to include species that in different time and space has resulted in considerable produce solitary conidia from conidiophores that may be difficulties in developing a stable phylogenetic classifica- simple or branched. This generic delineation based on a tion system in kingdom Fungi (Shenoy et al. 2007). This single anamorphic character has been accepted by several has partially resulted from the establishment of the dual authors (e.g. Kuthubutheen and Nawawi 1991;Bhatand system of classification in which sexual and asexual fungal Kendrick 1993;GohandHyde1996, 1998;Hoetal. forms are grouped separately. In this arrangement, more 2002). than 20,000 anamorphs have been grouped in a pigeon- Information on teleomorphs of Diplococcium and Spadi- hole-like system of genera, mainly based on anamorphic coides members is limited and ambiguous. Available taxo- characters (Shenoy et al. 2007). The practice of a dual nomic information from morphological and culture-based classification goes against the principle of natural classifi- studies suggests that Diplococcium species might have cation. Anamorphic genera are regularly described by connections with phylogenetically divergent sexual morphol- fungal taxonomists to facilitate practical purposes such as ogies. The suggested sexual forms include Helminthosphaeria identification, but with little consideration for the principle (Sordariomycetes)(Bisby1938; Ellis 1971b; Sutton 1973; of natural groupings in classification. Recent efforts in Subramanian 1983; Samuels et al. 1997; Goh and Hyde 1998; fungal gene sequence-data analysis have shown that many Réblová 1999b)andOtthia (Dothideomycetes)(Subramanian ‘well-established’ anamorphic genera are polyphyletic (e.g. and Sekar 1989). There is one report that, based on Seifert et al. 2000; Shenoy et al. 2006, 2007; Crous 2009; circumstantial evidence such as growth of the anamorph Crous et al. 2009b; Jones et al. 2009). There is a warranted on ascomata, suggested Tengiomyces as a possible sexual need to revisit the taxonomy of anamorphic genera and form of Spadicoides (Réblová 1999a). These reports refine their taxonomy based on a polyphasic approach as indicate that members of Diplococcium-Spadicoides are demonstrated in recent studies (Groenewald et al. 2008; possibly derived from phylogenetically distant lineages. Aveskamp et al. 2008, 2009, 2010). This paper presents our Phylogenetic significance of conidial catenation in Diplococ- recent studies on molecular phylogeny of seven anamorphic cium-Spadicoides taxonomy is, therefore, doubtful and genera that include members of the Diplococcium-complex questionable. DNA sequence-analysis is employed in this viz. Diplococcium Grove (1885), Spadicoides S. Hughes study to determine the phylogenetic relationships and (1958), Paliphora Sivanesan and B. Sutton (1985), and possible taxonomic placement of these asexual fungi within Polyschema H.P. Upadhyay (1966) and three anamorphic known ascomycetes. genera reported from India viz. Bahusutrabeeja Subrama- Paliphora is similar to Spadicoides in conidial ontogeny nian and Bhat (1977), Natarajania Pratibha and Bhat but produces hyaline, euseptate conidia (Sivanesan and (2005) and Rattania Prabhugaonkar and Bhat (2009). Sutton 1985), whereas Polyschema differs from Spadi- Diplococcium and Spadicoides have similar conidial coides mainly in having monotretic or polytretic conidiog- ontogeny. In both genera conidiogenous cells are terminal enous cells borne on micronematous conidiophores or intercalary, and polytretic with several unthickened (Upadhyay 1966; Ellis 1971a;Hoetal.2002). Inclusion conidiogenous loci (pores). Conidial ontogeny is holoblas- of Paliphora and Polyschema in this study aims to examine tic and the conidiogenous pores are easily visible after whether morphological similarities between these two conidial secession. The conidia are acropleurogenous, dry, genera and Spadicoides correspond to the DNA-based dematiaceous, usually thick-walled, 0–5-septate and may phylogenetic groupings. This paper also deals with phylo- Fungal Diversity (2010) 44:161–169 163 genetic placement of three hyphomycetous genera, Bahusu- spanning approximately 900 bp for partial 28S rRNA trabeeja, Natarajania and Rattania reported from India. gene were checked on 1% agarose electrophoresis gel Bahusutrabeeja was described to accommodate a hypho- stained with ethidium bromide. PCR products were then mycetous fungus that produces unicellular, hyaline, pear- purified using commercial kits (Amersham Biosciences, shaped to subglobose phialoconidia with several evenly UK, catalogue number 27-09602-01; Qiagen, Valencia, distributed appendages (Subramanian and Bhat 1977). This USA, catalogue number 28706). DNA sequencing was species has been frequently reported from aquatic habitats performed using the above-mentioned primers in an in Hong Kong and Guang Dong areas of China (Tsui et al. Applied Biosystem 3130/3730 analyzer at the Genome 2001; Wu and McKenzie 2003). Recently, this species has Research Centre, The University of Hong Kong