Bernard Paul

Secretion of β-1,3-Glucanases by the Yeast Pichia membranifaciens and Its Possible Role in the Biocontrol of Botrytis cinerea Causing Grey Mold Disease of the Grapevine

Pichia membranifaciens strain FY-101, isolated from grape skin, was found to be antagonistic to Botrytis cinerea, the causal organism of the grey mold disease of the grapevine. When grown together on solid as well as in liquid media, the yeast brings about the inhibition of Botrytis cinerea, which in turn loses its ability to produce the grey mold symptoms on the grapevine plantlets. The secretion of β-1,3-glucanases by P. membranifaciens is one of the possible mechanisms related to this antagonism. In vitro experiments confirm that this yeast can be used as a biological control organism against B. cinerea. An account of this antagonism and the production of β-1,3-glucanases by P. membranifaciens is given here.

Intraspecific and within-isolate sequence variation in the ITS rRNA gene region of Pythium mercuriale sp. nov. (Pythiaceae)

Sixteen Pythium isolates from diverse hosts and locations, which showed similarities in their morphology and sequences of the internal transcribed spacer (ITS) region of their rRNA gene, were investigated. As opposed to the generally accepted view, within single isolates ITS sequence variations were consistently found mostly as part of a tract of identical bases (A-T) within ITS1, and of GT or GTTT repeats within the ITS2 sequence. Thirty-one different ITS sequences obtained from 39 cloned ITS products from the 16 isolates showed high sequence and length polymorphisms within and between isolates. However, in a phylogenetic analysis, they formed a cluster distinct from those of other Pythium species. Additional sequencing of two nuclear genes (elongation factor 1 alpha and beta-tubulin) and one mitochondrial gene (nadh1) revealed high levels of heterozygosity as well as polymorphism within and between isolates, with some isolates possessing two or more alleles for each of the nuclear genes. In contrast to the observed variation in the ITS and other gene areas, all isolates were phenotypically similar. Pythium mercuriale sp. nov. (Pythiaceae) is characterized by forming thin-walled chlamydospores, subglobose to obovoid, papillate sporangia proliferating internally and smooth-walled oogonia surrounded by multiple antheridia. Maximum likelihood phylogenetic analyses based on both ITS and beta-tubulin sequence data place P. mercuriale in a clade between Pythium and Phytophthora.

Pythium carbonicum, a new species isolated from a spoil heap in northern France, the ITS region, taxonomy and comparison with related species

Pythium carbonicum (F-72) sp. nov. was found in soil samples taken on the top of a spoil heap in northern France. The morphology of this new species resembles that of a recently described species: Pythium megacarpum. However, the antheridial and oogonial characteristics of this new species are unique, and the comparison of its ITS region of the nuclear ribosomal DNA indicates that this species is also related to the genus Phytophthora. The fungus does not sporulate, the sporangia germinate directly into mycelium through germ tubes. The oogonia of P. carbonicum are smooth-walled and also papillated, and are provided with monoclinous and diclinous antheridia that wrap around, forming a complicated knot. Morphological features of this new species, together with the sequences of the ITS region of its nuclear ribosomal DNA and its comparison with related species are discussed here.

Characterisation of a new species of Pythium isolated from a wheat field in northern France and its antagonism towards Botrytis cinerea causing the grey mould disease of the grapevine.

A new species, Pythium bifurcatum, isolated from soil samples taken from a wheat field in Lille in northern France is described here. The oomycete occurred thrice out of 50 samples. The type specimen is F-91, which is a slow-growing saprophyte living on vegetable debris and which can be recognised by its antheridial as well as oogonial characteristics, which are different from other known species of Pythium. When grown together with Botrytis cinerea, the causal agent of the grey mould disease of the grapevine, Pythium bifurcatum shows a pronounced antagonism and suppresses its growth. Morphological features of this new species, its antagonism to B. cinerea, the sequences of the ITS region of its nuclear ribosomal DNA, and its comparison with related species are discussed in this article.

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.

Pythium campanulatum sp. nov., isolated from the rhizosphere of maize, its taxonomy, ITS region of rDNA, and comparison with related species.

Pythium campanulatum sp. nov. was isolated from some soil samples taken in the rhizosphere of maize (Zea mays) in north-eastern India. This species is characterized by the absence of zoospores and sporangia, antheridial branches wrapping around the oogonia leaving one to two campanulate antheridial cells after fertilization, and aplerotic oospores. The ITS region of its rDNA is comprised of 922 bases. This oomycete is closely related to Pythium orthogonon, Pythium nunn and Pythium toruloides. However, it has its own characteristic features and is completely devoid of zoospores. 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.

Gnomoniopsis smithogilvyi causes chestnut canker symptoms in Castanea sativa shoots in Switzerland.

A screening of Castanea sativa scions for grafting for the presence of endophytes showed that the opportunistic fungal pathogen Gnomoniopsis smithogilvyi was the most abundant member of the endophytic flora. This fungus is known as a pathogen affecting chestnut fruits in Italy and Australia. Here, we present evidence that it causes cankers very similar to the ones due to Cryphonectria parasitica infection on twigs and scions of chestnut trees. We found natural infections of G. smithogilvyi in healthy grafted plants as well as in scions from chestnut trees. The identity of the fungus isolated from asymptomatic tissues was verified by applying Kochs postulates and corroborated by DNA sequencing of four different gene regions. In contrast to C. parasitica that appears on the bark as yellow to orange pycnidia, stromata and slimy twisted tendrils, G. smithogilvyi forms orange to red and black pycnidia, gray stromata and cream-colored to beige slimy twisted tendrils on the bark. These Swiss strains are closely related to G. smithogilvyi strains from Australia and from New Zealand, Gnomoniopsis sp. and Gnomoniopsis castanea from New Zealand, Italy, France and Switzerland. While the strains from Ticino are genetically very close to G. smithogilvyi and G. castanea from Italy, the differences between the strains from Ticino and Geneva suggest two different origins. The present study supports the hypothesis that a single species named G. smithogilvyi, which is known to be the agent of chestnut rot, also causes wood cankers on chestnut.

A new mycoparasite, Pythium lycopersicum, isolated in Isparta, Turkey: morphology, molecular characteristics, and its antagonism with phytopathogenic fungi

Pythium lycopersicum sp. nov. has been isolated from soil samples taken in an agricultural land in the Isparta region of Southern Turkey. This oomycete is characterized by its contiguous sporangia having globose to elongated elements linked with hyphal filaments, ornamented oogonia, and monoclinous antheridia with large antheridial cells. The oomycete is reminiscent of Pythium ornamentatum described by the corresponding author in 1987 from soil samples taken in Algeria. Sequence analyses of the internal transcribed spacer (ITS) regions of rRNA show a close relationship with Pythium oligandrum and other mycoparasites possessing ornamented oogonia. Morphological and molecular features of this isolate justify its description as a new species: P. lycopersicum. When grown together with Botrytis cinerea, this oomycete shows a pronounced antagonism and suppresses the phytopathogen. The morphological details, together with the analysis of the ITS region of its rRNA, and its antagonism with some phytopathogens are discussed in this article.

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.

Pythium glomeratum, a new species isolated from agricultural soil taken in north-eastern France, its ITS region and its comparison with related species.

Pythium glomeratum sp. nov. is described here. It was isolated from soil samples taken in the northern France in 1992, was wrongly identified as Pythium heterothallicum and was kept aside. Recently, the ITS region of the rDNA of this oomycete was amplified and sequenced. The differences between the sequence and a more detailed study of the morphological characters of these two species, revealed that both are related but different species. P. glomeratum is characterized by the presence of branched antheridia that wrap around the oogonia, aplerotic to almost plerotic oospores, and the lack of zoosporangia and zoospores. Taxonomical description of this new species, its comparison with related oomycetes, polymerase chain reaction (PCR) amplification of the internal transcribed region (spacers ITS1, ITS2, and the gene 5.8S) of its ribosomal nuclear DNA and the nucleotide sequence of this region are given here.