Novel Curvularia species from clinical specimens
Madrid, H. ; Cunha, K.C. da; Gené, J. ; Dijksterhuis, J. ; Cano, J. ; Sutton, D.A. ; Guarro, J. ; Crous, P.W. - \ 2014
Persoonia 33 (2014). - ISSN 0031-5850 - p. 48 - 60.
internal transcribed spacer - allergic bronchopulmonary mycosis - antifungal susceptibility - bipolaris-australiensis - invasive sinusitis - genus curvularia - ribosomal dna - alternaria - cochliobolus - helminthosporium
The fungal genus Curvularia includes numerous plant pathogens and some emerging opportunistic pathogens of humans. In a previous study we used morphology and sequences of the nuclear ribosomal internal transcribed spacer region (ITS) and the glyceraldehyde-3-phosphate dehydrogenase (gpd) gene to identify species within a set of 99 clinical Curvularia isolates from the USA. Seventy-two isolates could be identified while the remaining 27 isolates belonged in three unclassified clades that were tentatively labelled Curvularia sp. I, II and III. In the present study, we further assess the taxonomic placement of these isolates using sequences of ITS, gpd, the large subunit rDNA, and the second largest subunit of RNA polymerase II. DNA sequence comparisons with a set of 87 isolates representing 33 Curvularia spp. and members of the closely-related genera Bipolaris and Exserohilum revealed that Curvularia sp. I, II and III represent novel lineages in Curvularia. These lineages are morphologically different from the currently accepted species. In the phylogenetic tree, Curvularia sp. I and sp. III were each split into two distinct lineages. Morphology and phylogeny supported the proposal of five new species, to be named C. americana, C. chlamydospora, C. hominis, C. muehlenbeckiae and C. pseudolunata. The concatenated 4-locus phylogeny revealed the existence of six clades in Curvularia, which are associated with particular morphological features. They were named after representative species, namely americana, eragrostidis, hominis, lunata, spicifera and trifolii.
Finding needles in haystacks: linking scientific names, reference specimens and molecular data for Fungi
Schoch, C.L. ; Robbertse, B. ; Robert, V. ; Vu, D. ; Cardinali, G. ; Irinyi, L. ; Meyer, W. ; Nilsson, R.H. ; Hughes, K. ; Miller, A.N. ; Kirk, P.M. ; Abarenkov, K. ; Aime, M.C. ; Ariyawansa, H.A. ; Bidartondo, M. ; Boekhout, T. ; Buyck, B. ; Cai, Q. ; Chen, J. ; Crespo, A. ; Crous, P.W. ; Damm, U. ; Beer, Z.W. de; Dentinger, B.T.M. ; Divakar, P.K. ; Duenas, M. ; Feau, N. ; Fliegerova, K. ; Garcia, M.A. ; Ge, Z.W. ; Griffith, G.W. ; Groenewald, J.Z. ; Groenewald, M. ; Grube, M. ; Gryzenhout, M. ; Gueidan, C. ; Guo, L. ; Hambleton, S. ; Hamelin, R. ; Hansen, K. ; Hofstetter, V. ; Hong, S.B. ; Houbraken, J. ; Hyde, K.D. ; Inderbitzin, P. ; Johnston, P.A. ; Karunarathna, S.C. ; Koljalg, U. ; Kovacs, G.M. ; Kraichak, E. ; Krizsan, K. ; Kurtzman, C.P. ; Larsson, K.H. ; Leavitt, S. ; Letcher, P.M. ; Liimatainen, K. ; Liu, J.K. ; Lodge, D.J. ; Luangsa-ard, J.J. ; Lumbsch, H.T. ; Maharachchikumbura, S.S.N. ; Manamgoda, D. ; Martin, M.P. ; Minnis, A.M. ; Moncalvo, J.M. ; Mule, G. ; Nakasone, K.K. ; Niskanen, T. ; Olariaga, I. ; Papp, T. ; Petkovits, T. ; Pino-Bodas, R. ; Powell, M.J. ; Raja, H.A. ; Redecker, D. ; Sarmiento-Ramirez, J.M. ; Seifert, K.A. ; Shrestha, B. ; Stenroos, S. ; Stielow, B. ; Suh, S.O. ; Tanaka, K. ; Tedersoo, L. ; Telleria, M.T. ; Udayanga, D. ; Untereiner, W.A. ; Dieguez Uribeondo, J. ; Subbarao, K.V. ; Vagvolgyi, C. ; Visagie, C. ; Voigt, K. ; Walker, D.M. ; Weir, B.S. ; Weiss, M. ; Wijayawardene, N.N. ; Wingfield, M.J. ; Xu, J.P. ; Yang, Z.L. ; Zhang, N. ; Zhuang, W.Y. ; Federhen, S. - \ 2014
Database : the Journal of Biological Databases and Curation 2014 (2014). - ISSN 1758-0463 - 21 p.
internal transcribed spacer - arbuscular mycorrhizal fungi - ribosomal dna - interspecific hybridization - sequence analyses - species complex - identification - evolution - barcode - life
DNA phylogenetic comparisons have shown that morphology-based species recognition often underestimates fungal diversity. Therefore, the need for accurate DNA sequence data, tied to both correct taxonomic names and clearly annotated specimen data, has never been greater. Furthermore, the growing number of molecular ecology and microbiome projects using high-throughput sequencing require fast and effective methods for en masse species assignments. In this article, we focus on selecting and re-annotating a set of marker reference sequences that represent each currently accepted order of Fungi. The particular focus is on sequences from the internal transcribed spacer region in the nuclear ribosomal cistron, derived from type specimens and/or ex-type cultures. Re-annotated and verified sequences were deposited in a curated public database at the National Center for Biotechnology Information (NCBI), namely the RefSeq Targeted Loci (RTL) database, and will be visible during routine sequence similarity searches with NR_prefixed accession numbers. A set of standards and protocols is proposed to improve the data quality of new sequences, and we suggest how type and other reference sequences can be used to improve identification of Fungi.
Porocercospora seminalis gen. et comb. nov., the causal organism of buffalograss false smut
Amaradasa, B.S. ; Madrid, H. ; Groenewald, J.Z. ; Crous, P.W. ; Amundsen, K. - \ 2014
Mycologia 106 (2014)1. - ISSN 0027-5514 - p. 77 - 85.
ribosomal dna - mycosphaerella - cochliobolus - cercospora - complex
False smut caused by Cercospora seminalis is an important disease of buffalograss (Buchloë dactyloides) affecting seed production. The pathogen prevents normal caryopsis development and causes considerable yield loss and reduced seed germination. The current taxonomic placement of the false-smut causal pathogen in the genus Cercospora is incorrect based on its morphological characteristics and DNA phylogeny. In the present study the phylogenetic position of C. seminalis is clarified based on DNA sequence analysis of three loci namely the internal transcribed spacer (ITS) region, partial nuclear ribosomal large subunit (LSU) and partial sequences of the RNA polymerase II second largest subunit (RPB2). A collection of C. seminalis isolates was made from buffalograss sites near Lincoln, Nebraska. DNA sequence data indicated that Cercospora seminalis is phylogenetically close to but distinct from species of Bipolaris and Curvularia (Pleosporaceae, Pleosporales). Cercospora seminalis morphologically had unique characteristics, namely densely aggregated and repeatedly branched conidiophores arising from a brown stroma, monotretic conidiogenous cells with inconspicuous loci, and scolecosporous conidia with distosepta, and thickened, darkened hila. Porocercospora is introduced as a new genus to accommodate the buffalograss false-smut pathogen.
Phylogenetic circumscription of Arthrographis (Eremomycetaceae, Dothideomycetes)
Giraldo, A. ; Gené, J. ; Sutton, D.A. ; Madrid, H. ; Cano, J. ; Crous, P.W. ; Guarro, J. - \ 2014
Persoonia 32 (2014). - ISSN 0031-5850 - p. 102 - 114.
keratinophilic fungi - molecular-data - ribosomal dna - sp-nov - ascomycota - tubeufiaceae - systematics - arthropsis - evolution - revision
Numerous members of Ascomycota and Basidiomycota produce only poorly differentiated arthroconidial asexual morphs in culture. These arthroconidial fungi are grouped in genera where the asexual-sexual connections and their taxonomic circumscription are poorly known. In the present study we explored the phylogenetic relationships of two of these ascomycetous genera, Arthrographis and Arthropsis. Analysis of D1/D2 sequences of all species of both genera revealed that both are polyphyletic, with species being accommodated in different orders and classes. Because genetic variability was detected among reference strains and fresh isolates resembling the genus Arthrographis, we carried out a detailed phenotypic and phylogenetic analysis based on sequence data of the ITS region, actin and chitin synthase genes. Based on these results, four new species are recognised, namely Arthrographis chlamydospora, A. curvata, A. globosa and A. longispora. Arthrographis chlamydospora is distinguished by its cerebriform colonies, branched conidiophores, cuboid arthroconidia and terminal or intercalary globose to subglobose chlamydospores. Arthrographis curvata produced both sexual and asexual morphs, and is characterised by navicular ascospores and dimorphic conidia, namely cylindrical arthroconidia and curved, cashew-nut-shaped conidia formed laterally on vegetative hyphae. Arthrographis globosa produced membranous colonies, but is mainly characterised by doliiform to globose arthroconidia. Arthrographis longispora also produces membranous colonies, but has poorly differentiated conidiophores and long arthroconidia. Morphological variants are described for A. kalrae and our results also revealed that Eremomyces langeronii and A. kalrae, traditionally considered the sexual and asexual morphs of the same species, are not conspecific.
A phylogenetic re-evaluation of Phyllosticta (Botryosphaeriales)
Wikee, S. ; Lombard, L. ; Nakashima, C. ; Motohashi, K. ; Chukeatirote, E. ; Cheewangkoon, R. ; McKenzie, E.H.C. ; Hyde, K.D. ; Crous, P.W. - \ 2013
Studies in Mycology 76 (2013)1. - ISSN 0166-0616 - p. 1 - 29.
citrus black spot - ribosomal dna - south-africa - fungus - banana - identification - eucalyptus - anamorph - vaccinii - nuclear
Phyllosticta is a geographically widespread genus of plant pathogenic fungi with a diverse host range. This study redefines Phyllosticta, and shows that it clusters sister to the Botryosphaeriaceae (Botryosphaeriales, Dothideomycetes), for which the older family name Phyllostictaceae is resurrected. In moving to a unit nomenclature for fungi, the generic name Phyllosticta was chosen over Guignardia in previous studies, an approach that we support here. We use a multigene DNA dataset of the ITS, LSU, ACT, TEF and GPDH gene regions to investigate 129 isolates of Phyllosticta, representing about 170 species names, many of which are shown to be synonyms of the ubiquitous endophyte P. capitalensis. Based on the data generated here, 12 new species are introduced, while epitype and neotype specimens are designated for a further seven species. One species of interest is P. citrimaxima associated with tan spot of Citrus maxima fruit in Thailand, which adds a fifth species to the citrus black spot complex. Previous morphological studies lumped many taxa under single names that represent complexes. In spite of this Phyllosticta is a species-rich genus, and many of these taxa need to be recollected in order to resolve their phylogeny and taxonomy.
Sizing up Septoria
Quaedvlieg, W. ; Verkley, G.J.M. ; Shin, H.D. ; Barreto, R.W. ; Alfenas, A.C. ; Swart, W.J. ; Groenewald, J.Z. ; Crous, P.W. - \ 2013
Studies in Mycology 75 (2013). - ISSN 0166-0616 - p. 307 - 390.
molecular-data sets - systematic reappraisal - phylogenetic lineages - genus septoria - ribosomal dna - similar fungi - sooty blotch - south-africa - mycosphaerella - genera
Septoria represents a genus of plant pathogenic fungi with a wide geographic distribution, commonly associated with leaf spots and stem cankers of a broad range of plant hosts. A major aim of this study was to resolve the phylogenetic generic limits of Septoria, Stagonospora, and other related genera such as Sphaerulina, Phaeosphaeria and Phaeoseptoria using sequences of the the partial 28S nuclear ribosomal RNA and RPB2 genes of a large set of isolates. Based on these results Septoria is shown to be a distinct genus in the Mycosphaerellaceae, which has mycosphaerella-like sexual morphs. Several septoria-like species are now accommodated in Sphaerulina, a genus previously linked to this complex. Phaeosphaeria (based on P. oryzae) is shown to be congeneric with Phaeoseptoria (based on P. papayae), which is reduced to synonymy under the former. Depazea nodorum (causal agent of nodorum blotch of cereals) and Septoria avenae (causal agent of avenae blotch of barley and rye) are placed in a new genus, Parastagonospora, which is shown to be distinct from Stagonospora (based on S. paludosa) and Phaeosphaeria. Partial nucleotide sequence data for five gene loci, ITS, LSU, EF-1a, RPB2 and Btub were generated for all of these isolates. A total of 47 clades or genera were resolved, leading to the introduction of 14 new genera, 36 new species, and 19 new combinations
A new approach to species delimitation in Septoria
Verkley, G.J.M. ; Quaedvlieg, W. ; Shin, H.D. ; Crous, P.W. - \ 2013
Studies in Mycology 75 (2013)1. - ISSN 0166-0616 - p. 213 - 305.
molecular-data sets - ribosomal dna - rdna sequence - mycosphaerella - anamorphs - phylogeny - genus - morphology - eucalyptus - celery
Septoria is a large genus of asexual morphs of Ascomycota causing leaf spot diseases of many cultivated and wild plants. Host specificity has long been a decisive criterium in species delimitation in Septoria, mainly because of the paucity of useful morphological characters and the high level of variation therein. This study aimed at improving the species delimitation of Septoria by adopting a polyphasic approach, including multilocus DNA sequencing and morphological analyses on the natural substrate and in culture. To this end 365 cultures preserved in CBS, Utrecht, The Netherlands, among which many new isolates obtained from fresh field specimens were sequenced. Herbarium material including many types was also studied. Full descriptions of the morphology in planta and in vitro are provided for 57 species. DNA sequences were generated for seven loci, viz. nuclear ITS and (partial) LSU ribosomal RNA genes, RPB2, actin, calmodulin, Btub, and EF. The robust phylogeny inferred showed that the septoria-like fungi are distributed over three main clades, establishing the genera Septoria s. str., Sphaerulina, and Caryophylloseptoria gen. nov. Nine new combinations and one species, Sphaerulina tirolensis sp. nov. were proposed. It is demonstrated that some species have wider host ranges than expected, including hosts from more than one family. Septoria protearum, previously only associated with Proteaceae was found to be also associated with host plants from six additional families of phanerogams and cryptogams. To our knowledge this is the first study to provide DNA-based evidence that multiple family-associations occur for a single species in Septoria. The distribution of host families over the phylogenetic tree showed a highly dispersed pattern for 10 host plant families, providing new insight into the evolution of these fungi. It is concluded that trans-family host jumping is a major force driving the evolution of Septoria and Sphaerulina
Diaporthe: a genus of endophytic, saprobic and plant pathogenic fungi
Gomes, R.R. ; Glienke, C. ; Videira, S.I.R. ; Lombard, L. ; Groenewald, J.Z. ; Crous, P.W. - \ 2013
Persoonia 31 (2013). - ISSN 0031-5850 - p. 1 - 41.
internal transcribed spacer - south-africa - species concepts - sp-nov - coelomycete phomopsis - foeniculum-vulgare - multigene analysis - north-america - ribosomal dna - twig dieback
Diaporthe (Phomopsis) species have often been reported as plant pathogens, non-pathogenic endophytes or saprobes, commonly isolated from a wide range of hosts. The primary aim of the present study was to resolve the taxonomy and phylogeny of a large collection of Diaporthe species occurring on diverse hosts, either as pathogens, saprobes, or as harmless endophytes. In the present study we investigated 243 isolates using multilocus DNA sequence data. Analyses of the rDNA internal transcribed spacer (ITS1, 5.8S, ITS2) region, and partial translation elongation factor 1-alpha (TEF1), beta-tubulin (TUB), histone H3 (HIS) and calmodulin (CAL) genes resolved 95 clades. Fifteen new species are described, namely Diaporthe arengae, D. brasiliensis, D. endophytica, D. hongkongensis, D. inconspicua, D. infecunda, D. mayteni, D. neoarctii, D. oxe, D. paranensis, D. pseudomangiferae, D. pseudophoenicicola, D. raonikayaporum, D. schini and D. terebinthifolii. A further 14 new combinations are introduced in Diaporthe, and D. anacardii is epitypified. Although species of Diaporthe have in the past chiefly been distinguished based on host association, results of this study confirm several taxa to have wide host ranges, suggesting that they move freely among hosts, frequently co-colonising diseased or dead tissue. In contrast, some plant pathogenic and endophytic taxa appear to be strictly host specific. Given this diverse ecological behaviour among members of Diaporthe, future species descriptions lacking molecular data (at least ITS and HIS or TUB) should be strongly discouraged.
Woudenberg, J.H.C. ; Groenewald, J.Z. ; Binder, M. ; Crous, P.W. - \ 2013
Studies in Mycology 75 (2013)1. - ISSN 0166-0616 - p. 171 - 212.
ribosomal dna - phylogenetic-relationships - species-group - ulocladium - embellisia - genus - identification - sequences - taxonomy - nuclear
Alternaria is a ubiquitous fungal genus that includes saprobic, endophytic and pathogenic species associated with a wide variety of substrates. In recent years, DNA-based studies revealed multiple non-monophyletic genera within the Alternaria complex, and Alternaria species clades that do not always correlate to species-groups based on morphological characteristics. The Alternaria complex currently comprises nine genera and eight Alternaria sections. The aim of this study was to delineate phylogenetic lineages within Alternaria and allied genera based on nucleotide sequence data of parts of the 18S nrDNA, 28S nrDNA, ITS, GAPDH, RPB2 and TEF1-alpha gene regions. Our data reveal a Pleospora/Stemphylium clade sister to Embellisia annulata, and a well-supported Alternaria clade. The Alternaria clade contains 24 internal clades and six monotypic lineages, the assemblage of which we recognise as Alternaria. This puts the genera Allewia, Brachycladium, Chalastospora, Chmelia, Crivellia, Embellisia, Lewia, Nimbya, Sinomyces, Teretispora, Ulocladium, Undifilum and Ybotromyces in synonymy with Alternaria. In this study, we treat the 24 internal clades in the Alternaria complex as sections, which is a continuation of a recent proposal for the taxonomic treatment of lineages in Alternaria. Embellisia annulata is synonymised with Dendryphiella salina, and together with Dendryphiella arenariae, are placed in the new genus Paradendryphiella. The sexual genera Clathrospora and Comoclathris, which were previously associated with Alternaria, cluster within the Pleosporaceae, outside Alternaria s. str., whereas Alternariaster, a genus formerly seen as part of Alternaria, clusters within the Leptosphaeriaceae. Paradendryphiella is newly described, the generic circumscription of Alternaria is emended, and 32 new combinations and 10 new names are proposed. A further 10 names are resurrected, while descriptions are provided for 16 new Alternaria sections
Phylogeography of the Sponge Suberites diversicolor in Indonesia: Insights into the Evolution of Marine Lake Populations
Becking, L.E. ; Erpenbeck, D. ; Peijnenburg, K.T.C.A. ; Voogd, N.J. de - \ 2013
PLoS ONE 8 (2013)10. - ISSN 1932-6203
oxidase subunit-i - coral triangle - halocaridina-rubra - ribosomal dna - indo-pacific - life-history - porifera - biodiversity - islands - demospongiae
The existence of multiple independently derived populations in landlocked marine lakes provides an opportunity for fundamental research into the role of isolation in population divergence and speciation in marine taxa. Marine lakes are landlocked water bodies that maintain a marine character through narrow submarine connections to the sea and could be regarded as the marine equivalents of terrestrial islands. The sponge Suberites diversicolor (Porifera: Demospongiae: Suberitidae) is typical of marine lake habitats in the Indo-Australian Archipelago. Four molecular markers (two mitochondrial and two nuclear) were employed to study genetic structure of populations within and between marine lakes in Indonesia and three coastal locations in Indonesia, Singapore and Australia. Within populations of S. diversicolor two strongly divergent lineages (A & B) (COI: p = 0.4% and ITS: p = 7.3%) were found, that may constitute cryptic species. Lineage A only occurred in Kakaban lake (East Kalimantan), while lineage B was present in all sampled populations. Within lineage B, we found low levels of genetic diversity in lakes, though there was spatial genetic population structuring. The Australian population is genetically differentiated from the Indonesian populations. Within Indonesia we did not record an East-West barrier, which has frequently been reported for other marine invertebrates. Kakaban lake is the largest and most isolated marine lake in Indonesia and contains the highest genetic diversity with genetic variants not observed elsewhere. Kakaban lake may be an area where multiple putative refugia populations have come into secondary contact, resulting in high levels of genetic diversity and a high number of endemic species
Species concepts in Cercospora: spotting the weeds among the roses
Groenewald, J.Z. ; Nakashima, C. ; Nishikawa, J. ; Shin, H.D. ; Park, J.H. ; Jama, A.N. ; Groenewald, M. ; Braun, U. ; Crous, P.W. - \ 2013
Studies in Mycology 75 (2013). - ISSN 0166-0616 - p. 115 - 170.
leaf-spot - water hyacinth - phylogenetic analysis - sequence-analysis - ribosomal dna - primer sets - new-zealand - sugar-beet - mycosphaerella - beticola
The genus Cercospora contains numerous important plant pathogenic fungi from a diverse range of hosts. Most species of Cercospora are known only from their morphological characters in vivo. Although the genus contains more than 5 000 names, very few cultures and associated DNA sequence data are available. In this study, 360 Cercospora isolates, obtained from 161 host species, 49 host families and 39 countries, were used to compile a molecular phylogeny. Partial sequences were derived from the internal transcribed spacer regions and intervening 5.8S nrRNA, actin, calmodulin, histone H3 and translation elongation factor 1-alpha genes. The resulting phylogenetic clades were evaluated for application of existing species names and five novel species are introduced. Eleven species are epi-, lecto- or neotypified in this study. Although existing species names were available for several clades, it was not always possible to apply North American or European names to African or Asian strains and vice versa. Some species were found to be limited to a specific host genus, whereas others were isolated from a wide host range. No single locus was found to be the ideal DNA barcode gene for the genus, and species identification needs to be based on a combination of gene loci and morphological characters. Additional primers were developed to supplement those previously published for amplification of the loci used in this study.
Phenotypic, Molecular, and Pathological Characterization of Colletotrichum acutatum Associated with Andean Lupine and Tamarillo in the Ecuadorian Andes
Falconi, C. ; Visser, R.G.F. ; Heusden, A.W. van - \ 2013
Plant Disease 97 (2013)6. - ISSN 0191-2917 - p. 819 - 827.
phytophthora-infestans - phylogenetic-relationships - olive anthracnose - ribosomal dna - sensu-lato - strawberry - identification - diversity - gloeosporioides - tomato
Anthracnose is a serious problem of both Andean lupine and tamarillo in Ecuador. Morphological features, internal transcribed spacer (ITS) sequences, and host specificity were used to characterize Colletotrichum isolates from lupine and tamarillo. Based on phenotypic and molecular characterization, the causal agent of anthracnose on both hosts was Colletotrichum acutatum. All isolates were identified in a C. acutatum-specific polymerase chain reaction assay. Colony diameter, conidia shape, and insensitivity to benomyl also placed isolates from both hosts in the C. acutatum group. However, a detailed analysis of the ITS sequences placed the lupine and tamarillo isolates from the Ecuadorian Andean zone in two clades, with both lupine and tamarillo isolates in each clade. C. acutatum isolates from Andean lupine were distinct from other C. acutatum isolates on lupine around the world. In cross-infection studies, the diameter of lesions produced by isolates from each host was compared on the main stem of two tamarillo and three lupine cultivars. Some isolates produced larger lesions on the host from which they were isolated but others showed similar aggressiveness on their alternate host. Isolates from both hosts were biotrophic on lupine stems, producing little necrosis and abundant sporulation whereas, on tamarillo stems, they produced dark lesions with few conidia. The collection of C. acutatum isolates from lupine and tamarillo provides interesting material for the study quantitative host adaptation.
A Universal Microarray Detection Method for Identification of Multiple Phytophthora spp. Using Padlock Probes
Sikora, K. ; Verstappen, E.C.P. ; Mendes, O. ; Schoen, C.D. ; Ristaino, J. ; Bonants, P.J.M. - \ 2012
Phytopathology 102 (2012)6. - ISSN 0031-949X - p. 635 - 645.
polymerase-chain-reaction - internal transcribed spacer - real-time pcr - ribosomal dna - phylogenetic-relationships - natural ecosystems - plant-pathogens - reaction assay - ramorum - quantification
The genus Phytophthora consists of many species that cause important diseases in ornamental, agronomic, and forest ecosystems worldwide. Molecular methods have been developed for detection and identification of one or several species of Phytophthora in single or multiplex reactions. In this article, we describe a padlock probe (PLP)-based multiplex method of detection and identification for many Phytophthora spp. simultaneously. A generic TaqMan polymerase chain reaction assay, which detects all known Phytophthora spp., is conducted first, followed by a species-specific PLP ligation. A 96-well-based microarray platform with colorimetric readout is used to detect and identify the different Phytophthora spp. PLPs are long oligonucleotides containing target complementary sequence regions at both their 5' and 3' ends which can be ligated on the target into a circular molecule. The ligation is point mutation specific; therefore, closely related sequences can be differentiated. This circular molecule can then be detected on a microarray. We developed 23 PLPs to economically important Phytophthora spp. based upon internal transcribed spacer-1 sequence differences between individual Phytophthora spp. Tests on genomic DNA of many Phytophthora isolates and DNA from environmental samples showed the specificity and utility of PLPs for Phytophthora diagnostics.
How well do ITS rDNA sequences differentiate species of true morels (Morchella)?
Du, X.H. ; Zhao, Q. ; Yang, Z.L. ; Hansen, K. ; Taskin, H. ; Büyükalaca, S. ; Dewsbury, D. ; Moncalvo, J.M. ; Douhan, G.W. ; Robert, V.A.R.G. ; Crous, P.W. ; Rehner, S.A. ; Rooney, A.P. ; Sink, S. ; O’Donnell, K. - \ 2012
Mycologia 104 (2012)6. - ISSN 0027-5514 - p. 1351 - 1368.
dna-sequences - phylogenetic-relationships - intragenomic variation - kingdom fungi - ribosomal dna - diversity - fusarium - identification - databases - recognition
Arguably more mycophiles hunt true morels (Morchella) during their brief fruiting season each spring in the northern hemisphere than any other wild edible fungus. Concerns about overharvesting by individual collectors and commercial enterprises make it essential that science-based management practices and conservation policies are developed to ensure the sustainability of commercial harvests and to protect and preserve morel species diversity. Therefore, the primary objectives of the present study were to: (i) investigate the utility of the ITS rDNA locus for identifying Morchella species, using phylogenetic species previously inferred from multilocus DNA sequence data as a reference; and (ii) clarify insufficiently identified sequences and determine whether the named sequences in GenBank were identified correctly. To this end, we generated 553 Morchella ITS rDNA sequences and downloaded 312 additional ones generated by other researchers from GenBank using emerencia and analyzed them phylogenetically. Three major findings emerged: (i) ITS rDNA sequences were useful in identifying 48/62 (77.4%) of the known phylospecies; however, they failed to identify 12 of the 22 species within the species-rich Elata Subclade and two closely related species in the Esculenta Clade; (ii) at least 66% of the named Morchella sequences in GenBank are misidentified; and (iii) ITS rDNA sequences of up to six putatively novel Morchella species were represented in GenBank. Recognizing the need for a dedicated Web-accessible reference database to facilitate the rapid identification of known and novel species, we constructed Morchella MLST (http://www.cbs.knaw.nl/morchella/), which can be queried with ITS rDNA sequences and those of the four other genes used in our prior multilocus molecular systematic studies of this charismatic genus.
DNA barcoding of Mycosphaerella species of quarantine importance to Europe
Quaedvlieg, W. ; Groenewald, J.Z. ; Jesús Yáñez-Morales, M. de; Crous, P.W. - \ 2012
Persoonia 29 (2012). - ISSN 0031-5850 - p. 101 - 115.
eucalyptus leaves - ribosomal dna - leaf-spot - phylogeny - nuclear - fungus - genus - anamorphs - taxonomy - sequence
The EU 7th Framework Program provided funds for Quarantine Barcoding of Life (QBOL) to develop a quick, reliable and accurate DNA barcode-based diagnostic tool for selected species on the European and Mediterranean Plant Protection Organization (EPPO) A1/A2 quarantine lists. Seven nuclear genomic loci were evaluated to determine those best suited for identifying species of Mycosphaerella and/or its associated anamorphs. These genes included ß-tubulin (Btub), internal transcribed spacer regions of the nrDNA operon (ITS), 28S nrDNA (LSU), Actin (Act), Calmodulin (Cal), Translation elongation factor 1-alpha (EF-1a) and RNA polymerase II second largest subunit (RPB2). Loci were tested on their Kimura-2-parameter-based inter- and intraspecific variation, PCR amplification success rate and ability to distinguish between quarantine species and closely related taxa. Results showed that none of these loci was solely suited as a reliable barcoding locus for the tested fungi. A combination of a primary and secondary barcoding locus was found to compensate for individual weaknesses and provide reliable identification. A combination of ITS with either EF-1a or Btub was reliable as barcoding loci for EPPO A1/A2-listed Mycosphaerella species. Furthermore, Lecanosticta acicola was shown to represent a species complex, revealing two novel species described here, namely L. brevispora sp. nov. on Pinus sp. from Mexico and L. guatemalensis sp. nov. on Pinus oocarpa from Guatemala. Epitypes were also designated for L. acicola and L. longispora to resolve the genetic application of these names.
Stagonosporopsis spp. associated with ray blight disease of Asteraceae
Vaghefi, N. ; Pethybridge, S.J. ; Ford, R. ; Nicolas, M.E. ; Crous, P.W. ; Taylor, P.W.J. - \ 2012
Australasian Plant Pathology 41 (2012)6. - ISSN 0815-3191 - p. 675 - 686.
phoma-ligulicola - phylogenetic analyses - ribosomal dna - mixed models - host-range - pyrethrum - sequences - complex - genera - taxa
Ray blight disease of pyrethrum (Tanacetum cinerariifolium) is shown to be caused by more than one species of Stagonosporopsis. The Australian pathogen, previously identified as Phoma ligulicola var. inoxydabilis, represents a new species described as Stagonosporopsis tanaceti based on morphological characters and a five-gene phylogeny employing partial sequences of the actin, translation elongation factor 1-alpha, internal transcribed spacers and 5.8S of the nrDNA, 28S large subunit and beta-tubulin 2 gene sequences. Furthermore, the two varieties of Stagonosporopsis ligulicola are elevated to species level as S. chrysanthemi and S. inoxydabilis based on their DNA phylogeny and morphology.
Phyllosticta species associated with freckle disease of banana
Wong, M.H. ; Crous, P.W. ; Henderson, J. ; Groenewald, J.Z. ; Drenth, A. - \ 2012
Fungal Diversity 56 (2012)1. - ISSN 1560-2745 - p. 173 - 187.
citrus black spot - ribosomal dna - mycosphaerella - sequences - musarum - plants
The identity of the casual agent of freckle disease of banana was investigated. The pathogen is generally referred to in literature under its teleomorphic name, Guignardia musae, or that of its purported anamorph, Phyllosticta musarum. Based on morphological and molecular data from a global set of banana specimens, several species were found associated with freckle disease. Phyllosticta maculata (from Southeast Asia and Oceania) is introduced as a new name for Guignardia musae, and an epitype is designated from Australia. Phyllosticta musarum (from India and Thailand) is shown to represent a distinct species, and the name is fixed by designation of an epitype from India. Guignardia stevensii is confirmed as distinct species from Hawaii, while Guignardia musicola from northern Thailand is shown to contain different taxa and is regarded as nomen confusum. Phyllosticta cavendishii is described as a new, widely distributed species, appearing primarily on Cavendish, but also on non-Cavendish banana cultivars.
Chocolate spot of Eucalyptus
Cheewangkoon, R. ; Groenewald, J.Z. ; Hyde, K.D. ; To-anun, C. ; Crous, P.W. - \ 2012
Mycological Progress 11 (2012)2. - ISSN 1617-416X - p. 61 - 69.
leaf-inhabiting fungi - ribosomal dna - genera nova - heteroconium - proteaceae - anamorphs - culture
Chocolate Spot leaf disease of Eucalyptus is associated with several Heteroconium-like species of hyphomycetes that resemble Heteroconium s.str. in morphology. They differ, however, in their ecology, with the former being plant pathogenic, while Heteroconium s.str. is a genus of sooty moulds. Results of molecular analyses, inferred from DNA sequences of the large subunit (LSU) and internal transcribed spacers (ITS) region of nrDNA, delineated four Heteroconium-like species on Eucalyptus, namely H. eucalypti, H. kleinziense, Alysidiella parasitica, and one isolate resembling a novel species in a clade separate from the holotype of Heteroconium, H. citharexyli. Based on molecular phylogeny, morphology and ecology, the Heteroconium-like species associated with Chocolate Spot disease are reclassified in the genus Alysidiella, which is shown to have mycelium that is immersed in and superficial on the host tissue and conidiogenous cells that can have loci that are either inconspicuous or proliferating percurrently. Furthermore, conidiogenous cells can either occur solitary on hyphae, or be sporodochial, arranged on a weakly developed stroma, which further distinguishes Alysidiella from Heteroconium
A re-appraisal of Harknessia (Diaporthales), and the introduction of Harknessiaceae fam. nov.
Crous, P.W. ; Summerell, B.A. ; Shivas, R.G. ; Carnegie, A.J. ; Groenewald, J.Z. - \ 2012
Persoonia 28 (2012). - ISSN 0031-5850 - p. 49 - 65.
phylogenetic lineages - ribosomal dna - genera nova - primer sets - eucalyptus - wuestneia - morphology - culture - fungi - genus
Harknessiaceae is introduced as a new family in the ascomycete order Diaporthales to accommodate species of Harknessia with their Wuestneia-like teleomorphs. The family is distinguished by having pycnidial conidiomata with brown, furfuraceous margins, brown conidia with hyaline, tube-like basal appendages, longitudinal striations, and rhexolytic secession. Six species occurring on Eucalyptus are newly introduced, namely H. australiensis, H. ellipsoidea, H. pseudohawaiiensis, and H. ravenstreetina from Australia, H. kleinzeeina from South Africa, and H. viterboensis from Italy. Epitypes are designated for H. spermatoidea and H. weresubiae, both also occurring on Eucalyptus. Members of Harknessia are commonly associated with leaf spots, but also occur as saprobes and endophytes in leaves and twigs of various angiosperm hosts.
Genera of diaporthalean coelomycetes associated with leaf spots of tree hosts
Crous, P.W. ; Summerell, B.A. ; Alfenas, A.C. ; Edwards, J. ; Pascoe, I.G. ; Porter, I.J. ; Groenewald, J.Z. - \ 2012
Persoonia 28 (2012). - ISSN 0031-5850 - p. 66 - 75.
phylogenetic lineages - ribosomal dna - south-africa - primer sets - eucalyptus - phaeoacremonium - grapevines - harknessia - culture - genus
Four different genera of diaporthalean coelomycetous fungi associated with leaf spots of tree hosts are morphologically treated and phylogenetically compared based on the DNA sequence data of the large subunit nuclear ribosomal DNA gene (LSU) and the internal transcribed spacers and 5.8S rRNA gene of the nrDNA operon. These include two new Australian genera, namely Auratiopycnidiella, proposed for a leaf spotting fungus occurring on Tristaniopsis laurina in New South Wales, and Disculoides, proposed for two species occurring on leaf spots of Eucalyptus leaves in Victoria. Two new species are described in Aurantiosacculus, a hitherto monotypic genus associated with leaf spots of Eucalyptus in Australia, namely A. acutatus on E. viminalis, and A. eucalyptorum on E. globulus, both occurring in Tasmania. Lastly, an epitype specimen is designated for Erythrogloeum hymenaeae, the type species of the genus Erythrogloeum, and causal agent of a prominent leaf spot disease on Hymenaea courbaril in South America. All four genera are shown to be allied to Diaporthales, although only Aurantiosacculus (Cryphonectriaceae) could be resolved to family level, the rest being incertae sedis.