DNA barcoding of oomycetes with cytochrome c oxidase subunit I and internal transcribed spacer.

Oomycete species occupy many different environments and many ecological niches. The genera Phytophthora and Pythium for example, contain many plant pathogens which cause enormous damage to a wide range of plant species. Proper identification to the species level is a critical first step in any investigation of oomycetes, whether it is research driven or compelled by the need for rapid and accurate diagnostics during a pathogen outbreak. The use of DNA for oomycete species identification is well established, but DNA barcoding with cytochrome c oxidase subunit I (COI) is a relatively new approach that has yet to be assessed over a significant sample of oomycete genera. In this study we have sequenced COI, from 1205 isolates representing 23 genera. A comparison to internal transcribed spacer (ITS) sequences from the same isolates showed that COI identification is a practical option; complementary because it uses the mitochondrial genome instead of nuclear DNA. In some cases COI was more discriminative than ITS at the species level. This is in contrast to the large ribosomal subunit, which showed poor species resolution when sequenced from a subset of the isolates used in this study. The results described in this paper indicate that COI sequencing and the dataset generated are a valuable addition to the currently available oomycete taxonomy resources, and that both COI, the default DNA barcode supported by GenBank, and ITS, the de facto barcode accepted by the oomycete and mycology community, are acceptable and complementary DNA barcodes to be used for identification of oomycetes.

A new species of Pythium with inflated sporangia and coiled antheridia, isolated from India.

Pythium kashmirense sp. nov. was isolated from soil samples taken on the Himalayas at the height of 5300 feet in the Shivalik Hill Range of the northern Indian state of Jammu and Kashmir. The oomycete has filamentous-inflated type sporangia and its antheridial filaments form loose loops around the female gametangia, and coil around the oogonial stalks. The new species is closely related to Pythium plurisporium, Pythium periilum, Pythium inflatum, and Pythium folliculosum. All of these oomycetes have filamentous-inflated type sporangia. However, P. kashmirense has its own distinguishing characteristics which can easily differentiate it among these related species. The sequences of the internal transcribed spacer (ITS) region of its rRNA and its morphological characters are unique for the genus Pythium. Taxonomic description of this new species, its comparison with related oomycetes and the sequence of the ITS region of its rRNA, are discussed here.

Saprolegnia bulbosa sp. nov. isolated from an Argentine stream: taxonomy and comparison with related species.

Saprolegnia bulbosa sp. nov. was isolated from floating and decaying twigs and leaves in El Gato stream, Partido de La Plata, Buenos Aires Province, Argentina. The distinctive characteristics of S. bulbosa are the product of smooth oogonia and predominantly contorted monoclinous, androgynous and diclinous antheridia. The oogonial stalks are usually bent, curved or once coiled; oospores are subcentric, (1) 2-15 (45) per oogonium and are variable in size. Taxonomical description of this new species, its comparison with related oomycetes of the genus and the nucleotide sequences of the internal transcribed region (spacers ITS1, ITS2 and the gene 5.8S) of its rRNA gene are given here.

Pythium rhizo-oryzae sp. nov. isolated from paddy fields: taxonomy, ITS region of rDNA, and comparison with related species.

Pythium rhizo-oryzae sp. nov. was isolated from some soil samples taken in paddy fields in northeastern India. For quite a long time it was wrongly identified as Pythium catenulatum Mathews, because of the presence of numerous catenulate hyphal bodies. The sequencing of the ITS region of its nuclear ribosomal DNA and a close look at its morphological characters have now enabled us to describe it as a new species. This species is characterized by the absence of zoospores and sporangia, rarely formed aritheridia and oogonia, and the presence of chained and terminal hyphal bodies. The ITS region of its rDNA comprises 774 bases. This oomycete is closely related to the members producing a filamentous inflated type of sporangia like P. catenulatum Mathews, P. torulosum Coker & Paterson, and P. inflatum Mathews. The closest of all these is P. catenulatum. Taxonomic description of this new species and its comparison with related oomycetes, together with the sequence of the PCR amplified internal transcribed region (spacers ITS1, ITS2, and the gene 5.8S) of its rDNA are given here.

A new species of Pythium with ornamented oogonia: morphology, taxonomy, internal transcribed spacer region of its ribosomal RNA, and its comparison with related species.

Pythium spiculum sp. nov. was isolated from soil samples taken in a vineyard in the Burgundian region of France and from different locations in Spain and Portugal. The oomycete has spiny oogonia and does not sporulate readily. It resembles Pythium mamillatum Meurs, but has its own distinguishing characteristics. It also exhibits sickle-shaped as well as spherical appressoria which at times are associated with sex organs like those found in Pythium abappressorium Paulitz and Pythium contiguanum Paul. Sequencing of the internal transcribed spacer region of its nuclear ribosomal DNA and a close look at its morphological characters have now enabled us to describe it as a new species. The internal transcribed spacer region of its rRNA gene sequence is comprised of 945 bases. This oomycete is closely related to the members that form ornamented or spiny oogonia like Pythiummamillatum, Pythium spinosum and Pythium irregulare but also with those producing smooth-walled oogonia like Pythium paroecandrum, Pythium sylvaticum and Pythium cylindrosporum. Taxonomic description of this new species, its comparison with related oomycetes, the sequence of the internal transcribed spacer region of its rRNA gene and the phylogenetic tree, are given here.

Morphological and molecular taxonomy of Pythium longisporangium sp. nov. isolated from the Burgundian region of France.

During the course of an investigation on the Pythiaceous oomycetes occurring in the Burgundian vineyards, some species of Pythium possessing mainly hypogynous antheridia were found. These had been classified as oomycetes belonging to the "Pythium rostratum" group for a long time. Three of these isolates, having similar structures and growth, are very closely related to a recently described species, Pythium bifurcatum Paul. A close look at these, however, underlines some fundamental differences with the latter. Not all of them produce zoospores but have very large sporangia. The type specimen is F-1200 (B 76a) which is a medium-slow growing saprophyte. The sequence of the ITS region of the rDNA also shows a very close relationship with P. bifurcatum. On the basis of morphological and molecular analysis, we now describe this species as Pythium longisporangium sp. nov. Morphological features of this new species, the sequences of the ITS region of its nuclear ribosomal DNA, and its comparison with related species are discussed.

Characterisation of Pythium paroecandrum and its antagonism towards Botrytis cinerea, the causative agent of grey mould disease of grape.

Pythium paroecandrum (B-30), an oomycete, was isolated from soil samples taken from a wheat field in Genlis in the Burgundy region of France and was found to check the growth and development of Botrytis cinerea, a serious grapevine pathogen. The oomycete is a fast-growing organism, living on vegetable debris, and can be recognised by its catenulate hyphal swellings, catenulate oogonia, and monoclinous antheridia. When grown together with B. cinerea, the causal agent of the grey mould disease of the grapevine, P. paroecandrum shows a pronounced antagonism and suppresses its growth and its aptitude to provoke the grey mould symptoms. Morphological features of this oomycete, its antagonism to B. cinerea, the sequences of the internal transcribed spacer region of its nuclear ribosomal DNA, and its comparison with related species are discussed in this article.