Highlights of the Didymellaceae: A polyphasic approach to characterise Phoma and related pleosporalean genera

Fungal taxonomists routinely encounter problems when dealing with asexual fungal species due to poly- and paraphyletic generic phylogenies, and unclear species boundaries. These problems are aptly illustrated in the genus Phoma. This phytopathologically significant fungal genus is currently subdivided into nine sections which are mainly based on a single or just a few morphological characters. However, this subdivision is ambiguous as several of the section-specific characters can occur within a single species. In addition, many teleomorph genera have been linked to Phoma, three of which are recognised here. In this study it is attempted to delineate generic boundaries, and to come to a generic circumscription which is more correct from an evolutionary point of view by means of multilocus sequence typing. Therefore, multiple analyses were conducted utilising sequences obtained from 28S nrDNA (Large Subunit - LSU), 18S nrDNA (Small Subunit - SSU), the Internal Transcribed Spacer regions 1 & 2 and 5.8S nrDNA (ITS), and part of the ß-tubulin (TUB) gene region. A total of 324 strains were included in the analyses of which most belonged to Phoma taxa, whilst 54 to related pleosporalean fungi. In total, 206 taxa were investigated, of which 159 are known to have affinities to Phoma. The phylogenetic analysis revealed that the current Boeremaean subdivision is incorrect from an evolutionary point of view, revealing the genus to be highly polyphyletic. Phoma species are retrieved in six distinct clades within the Pleosporales, and appear to reside in different families. The majority of the species, however, including the generic type, clustered in a recently established family, Didymellaceae. In the second part of this study, the phylogenetic variation of the species and varieties in this clade was further assessed. Next to the genus Didymella, which is considered to be the sole teleomorph of Phoma s. str., we also retrieved taxa belonging to the teleomorph genera Leptosphaerulina and Macroventuria in this clade. Based on the sequence data obtained, the Didymellaceae segregate into at least 18 distinct clusters, of which many can be associated with several specific taxonomic characters. Four of these clusters were defined well enough by means of phylogeny and morphology, so that the associated taxa could be transferred to separate genera. Aditionally, this study addresses the taxonomic description of eight species and two varieties that are novel to science, and the recombination of 61 additional taxa

Systematic reappraisal of species in Phoma section Paraphoma, Pyrenochaeta and Pleurophoma

Sequence data from the 18S nrDNA (SSU) and 28S nrDNA (LSU) regions of isolates of Phoma section Paraphoma were compared with those of representative isolates of the morphologically similar anamorph genera Pleurophoma and Pyrenochaeta and of the type species of Phoma sections Phoma, Pilosa and Plenodomus. Phoma section Paraphoma was found to be highly polyphyletic within the Pleosporales and only distantly related to Phoma section Phoma. The genus Paraphoma, which is based on Paraphoma radicina, is reintroduced in the Phaeosphaeriaceae with two additional taxa. The new genera Setophoma and Neosetophoma, type species Setophoma terrestris comb. nov. and Neosetophoma samarorum comb. nov., are introduced and represent species that are closely related to Paraphoma but differ based on morphological characters and molecular phylogeny. Phoma coonsii is transferred to genus Chaetosphaeronema that also belongs to the Phaeosphaeriaceae. Pyrenochaetopsis gen. nov. is introduced to accommodate the type species Pyrenochaetopsis leptospora comb. nov., as well as several other species formerly accommodated in Phoma and Pyrenochaeta. Pyrenochaetopsis is closely related to Pyrenochaeta and classified in the Cucurbitariaceae. Pleurophoma cava is transferred to genus Pyrenochaeta. The new genera elucidate the confusing taxonomy of species in genera Phoma, Pyrenochaeta and Pleurophoma and recognize monophyletic genera with distinct teleomorph affinities

Phyllosticta citriasiana sp. nov., the cause of Citrus tan spot of Citrus maxima in Asia

Guignardia citricarpa, the causal agent of Citrus Black Spot, is subject to phytosanitary legislation in the European Union and the U.S.A. This species is frequently confused with G. mangiferae, which is a non-pathogenic, and is commonly isolated as an endophyte from citrus fruits and a wide range of other hosts. Recently, necrotic spots similar to those caused by G. citricarpa were observed on fruit of Citrus maxima intercepted in consignments exported from Asia. In these spots, pycnidia and conidia of a Guignardia species closely resembling G. citricarpa were observed, and therefore measures were taken for the consignments in line with the European Union legislation for G. citricarpa. To determine the identity of the causal organism on this new host, fungal isolates were subjected to DNA sequence analysis of the internal transcribed spacer region (ITS1, 5.8S, ITS2), translation elongation factor 1-alpha (TEF1) and actin genes. A combined phylogenetic tree resolved three species correlating to G. citricarpa, G. mangiferae and a previously undescribed species, Phyllosticta citriasiana sp. nov., closely related to G. citricarpa. Morphologically P. citriasiana can be distinguished from G. citricarpa by having larger conidia, longer conidial appendages, and in not producing any diffuse yellow pigment when cultivated on oatmeal agar (OA). Furthermore, it is distinguishable from G. mangiferae by having smaller conidia, with a narrower mucoid sheath. In culture, colonies of P. citriasiana can also be distinguished from G. citricarpa and G. mangiferae by being darker shades of grey and black on OA, malt extract agar (MEA), potato-dextrose agar, and cornmeal agar. Furthermore, cultures of P. citriasiana achieved optimal growth after 2 weeks at 21-27°C, and ceased to grow at 30-33°C. In contrast, colonies of G. citricarpa and G. mangiferae achieved optimal growth at 27-30°C, and ceased to grow at 30-36°C Colonies of P. citriasiana also grew faster than those of G. citricarpa and G. mangiferae on OA and MEA. Phyllosticta citriasiana appears to be a harmful pathogen of Citrus maxima, causing a tan spot on fruit, underlining the need for further surveys and research to determine its distribution and host range

Regulatory Structures and their Impact on the Sustainability Performance of Public Transport in World Cities

This paper describes a new measure of the sustainability performance of public transport in 88 world cities adopting 15 indicators including Environmental, Social, Economic and System Effectiveness sustainability. Sustainability performance is then explored for cities with only “Public” operations or others with some degree of commercial operation (“Non-Public”) Results show no significant difference in aggregate total sustainability indicator scores between world cities with “Public”/“Non-Public” operations. However Social Sustainability indicators are significantly different with “Public” operations having better Social Sustainability performance than “Non-Public”. For individual component indicators, three of the four Social Sustainability component indicators have average normalised scores suggesting statistically significant differences between “Public” and “Non-Public” city scores with “Public” cities performing better than “Non-Public”. The indicators and their relative advantage to “Public” cities being Trip distance (24%), Affordability (34%) and PT related deaths (29%). However results also show that operating costs per passenger km and cost recovery are higher in “Non-Public” cities suggesting higher elements of Economic Sustainability in “Non-Public” based Public Transport cities The paper concludes with a summary and discussion of the results including implications for regulatory practices and areas for future research.Institute of Transport and Logistics Studies. Faculty of Economics and Business. The University of Sydne

Multi-locus phylogeny of Pleosporales: a taxonomic, ecological and evolutionary re-evaluation

Five loci, nucSSU, nucLSU rDNA, TEF1, RPB1 and RPB2, are used for analysing 129 pleosporalean taxa representing 59 genera and 15 families in the current classification of Pleosporales. The suborder Pleosporineae is emended to include four families, viz. Didymellaceae, Leptosphaeriaceae, Phaeosphaeriaceae and Pleosporaceae. In addition, two new families are introduced, i.e. Amniculicolaceae and Lentitheciaceae. Pleomassariaceae is treated as a synonym of Melanommataceae, and new circumscriptions of Lophiostomataceae s. str, Massarinaceae and Lophiotrema are proposed. Familial positions of Entodesmium and Setomelanomma in Phaeosphaeriaceae, Neophaeosphaeria in Leptosphaeriaceae, Leptosphaerulina, Macroventuria and Platychora in Didymellaceae, Pleomassaria in Melanommataceae and Bimuria, Didymocrea, Karstenula and Paraphaeosphaeria in Montagnulaceae are clarified. Both ecological and morphological characters show varying degrees of phylogenetic significance. Pleosporales is most likely derived from a saprobic ancestor with fissitunicate asci containing conspicuous ocular chambers and apical rings. Nutritional shifts in Pleosporales likely occured from saprotrophic to hemibiotrophic or biotrophic

Multiple Didymella teleomorphs are linked to the Phoma clematidina morphotype

The fungal pathogen Phoma clematidina is used as a biological agent to control the invasive plant species Clematis vitalba in New Zealand. Research conducted on P. clematidina as a potential biocontrol agent against C. vitalba, led to the discovery of two perithecial-forming strains. To assess the diversity of P. clematidina and to clarify the teleomorph-anamorph relationship, phylogenetic analyses of 18 P. clematidina strains, reference strains representing the Phoma sections in the Didymellaceae and strains of related species associated with Clematis were conducted. Partial sequences of the ITS1, ITS2 and 5.8S rRNA gene, the ß-tubulin gene and 28S rRNA gene were used to clarify intra- and inter-species relationships. These analyses revealed that P. clematidina resolves into three well-supported clades which appear to be linked to differences in host specificity. Based on these findings, Didymella clematidis is newly described and the descriptions of P. clematidina and D. vitalbina are amended

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