Biochemical characterisation of Pythium spp. involved in cavity spot of carrots in France

The pathogenicity and growth rate in vivo were assessed on 27 isolates of Pythium spp. recovered from cavity spot lesions on carrots grown in various parts of northwest France. Polyacrylamide gel electrophoresis of isoesterases was used to identify the Pythium spp. involved. Slow-growing isolates were more aggressive than fast-growing ones when inoculated on carrot tap roots. Isoesterase patterns identified the slow-growing isolates as P. violae and P. sulcatum; P. ultimum and P. intermedium were identified among the less aggressive fast-growing isolate group, in which some isolates were also classed as P. sylvaticum or P. irregulare, which have similar electrophoretic profiles. The incidence of Pythium spp. associated with the disease in France is discussed in regard to cavity spot in other countries.     

Compatibility of systemic acquired resistance and microbial biocontrol for suppression of plant disease in a laboratory assay

Systemic acquired resistance (SAR) and microbial biocontrol each hold promise as alternatives to pesticides for control of plant diseases. SAR and Bacillus cereus UW85, a microbial biocontrol agent, separately suppress seedling damping-off diseases caused by oomycete pathogens. The purposes of this study were to investigate how expression of SAR affected the efficacy of biocontrol by UW85 and if UW85 treatment of plants induced SAR. We devised a laboratory assay in which seedling damping-off disease, induction of SAR, and growth of UW85 could be quantified. Seedlings of Nicotiana tabacum Xanthi nc were germinated on moist filter paper and transferred after 7 days to water agar plates (40 seedlings per plate). Zoospores of oomycete pathogens (Pythium torulosum, Pythium aphanidermatum, or Phytophthora parasitica) were applied at concentrations that caused 80% seedling mortality within 10 days. Seedling mortality was dependent on zoospore inoculum concentration. The level of disease suppression caused by treatment with UW85 depended on the UW85 dose applied. SAR was induced with 0.5-mM salicylic acid or 0.1-mM 2,6-dichloroisonicotinic acid. Expression of an SAR-related gene was confirmed by northern analysis with a probe prepared from a tobacco PR-1a cDNA. Induction of SAR suppressed disease caused by each of the oomycete pathogens, but did not alter the growth of UW85 on roots. Treatment of seedlings with UW85 did not induce the expression of PR-1a. The combination of induction of SAR and treatment with UW85 resulted in additive suppression of disease as measured by seedling survival.     

Fungicidal drenches for control of root rot in subterranean clover

Fungicide drenches of benomyl, metalaxyl, iprodione, propamocarb, or thiram were applied to intact soil cores taken from known root rot affected fields in Western Australia, to control subterranean clover root rot. Metalaxyl was the most effective in reducing seedling damping-off. The most effective fungicide for reducing the level of rotting of both tap and lateral root systems of survixing plants varied from season to season at one site and varied between different sites in any one season with each fungicide giving a significant reduction in root disease on at least one occasion. Results suggest that different individuals or complexes of root pathogens were operative between seasons in any one site, and between sites for any one season. In some instances it appears that different individual root pathogens or pathogen complexes were operative on tap roots compared to lateral roots.     

Elicitor-induced cell death and phytoalexin synthesis in Daucus carota L.

Suspension-cultured carrot cells and intact leaves respond to crude and purified protein elicitors from the non-host fungus Pythium aphanidermatum by activating the general phenylpropanoid pathway and incorporating de-novo-synthesized 4-hydroxybenzoic acid (4-HBA) into the cell wall. The cultured cells undergo a very rapid elicitor-induced cell death. Both reactions are directly correlated in their time course and their dose dependency. Cell death in elicitor-treated protoplasts resulted in early membrane damage and the digestion of DNA into oligonucleosomal fragments. The same pattern of DNA degradation could be induced in protoplasts by the G-protein activators Mas-7 or mastoparan. In cell cultures, both activators induced a rapid loss of viability without the activation of the general phenylpropanoid pathway. The elicitor-induced reactions, the loss of viability and the induction of 4-HBA biosynthesis were blocked by the calcium-channel blocker nifedipine. Neomycin and U73122, two inhibitors of phospholipase C, blocked the induction of 4-HBA biosynthesis but did not affect the loss in viability. The injection of the elicitor into the leaves of intact carrot plants confirmed the results obtained with cell cultures with regard to the induction of the hypersensitive response. The purification of the active compound revealed a 25-kDa protein which triggers both cell death and 4-HBA synthesis. The signalling pathways to both reactions could be independently blocked or induced. Received: 27 February 1998 / Accepted: 25 May 1998     

E‐ADA activity in lymphocytes of an experimental model of pythiosis treated with immunotherapy

Pythiosis is a life‐threatening disease caused by the oomycete Pythium insidiosum. Some authors have suggested the involvement of a Th2‐like immune response in the infected host, which leads to extensive tissue damage. The switch from a Th2 to a Th1 response pattern is one hypothesis to explain the curative properties of immunotherapy. Taking into account the importance of immunotherapy for pythiosis treatment and the contribution of adenine nucleotides in the immunoregulation of the host, we evaluated the ecto‐adenosine deaminase (E‐ADA; EC 3·5.4·4) activity in lymphocytes from rabbits inoculated with P. insidiosum. Rabbits were inoculated with 1 milliliter of zoospores subcutaneously injected into the lateral thorax; after developing lesions, the rabbits received eight doses of immunotherapy. E‐ADA activity was measured in lymphocytes and the adenine nucleotides and adenosine levels were quantitatively determined in serum. Rabbits with characteristic lesions of pythiosis showed a decreased E‐ADA activity (82·36%), a decreased adenosine triphosphate concentration (54·04%) and a higher adenosine concentration (2·51 fold), when compared with controls, after 28 days of inoculation. However, after the immunotherapy, the rabbits showed an increase in the E‐ADA activity when compared with control (78·62%), contributing for the change in the immune response. Our results reinforce the hypothesis that the change from a Th2 to a Th1 immune response with the participation of the purinergic system could be responsible for the curative properties of immunotherapy. Copyright © 2012 John Wiley & Sons, Ltd.     

Pythium burgundicum sp. nov. isolated from soil samples taken in French vineyards

During the course of investigation on pythiaceous fungi occurring in the Burgundian vineyards, a new species of Pythium has been isolated. This oomycete is characterized by its nonproliferating and nonsporulating, spherical to cylindrical type of sporangia (hyphal bodies) germinating through germ tubes, smooth-walled oogonia that are supplied with hypogynous, monoclinous or rarely diclinous antheridia, and smooth-walled oospores. The antheridial cells are very prominent and are reminiscent of Pythium bifurcatum, Pythium segnitium and Pythium longandrum described previously by the author. The internal transcribed spacer region of the rRNA of this new species is composed of 883 bases, which is quite different from that of the closest relatives. Morphological and molecular features of this new species, named as Pythium burgundicum , are discussed in this article.     

Managing soilborne diseases of vegetable crops with a pre-plant soil or substrate amendment of a corn distillation product

The efficacy of condensed distiller's solubles (CDS), a co-product of ethanol production from corn, rich in organic matter, and high in carbon to nitrogen ratio, was tested as a pre-plant amendment against Verticillium wilt of eggplant and potato scab in potato soils from commercial fields and against damping-off diseases of radish and cucumber seedlings in a peat-based mix and muck soil. Eggplants grown in a potato soil amended with CDS (1% mass/mass) showed less Verticillium wilt and increased fresh (37-54%) and dry (31-45%) plant biomass compared to the control in the greenhouse. In a potato soil with medium levels of disease, CDS (1 or 2%) increased the percentage of marketable tubers by 116% under greenhouse, 119% under micro-plot and 75% under field conditions. In the growth room, CDS (1, 2, and 4%) amendment to a peat-based mix infested with Rhizoctonia solani 1 week before planting seeds improved the percentage of healthy radish seedlings (22-72% healthy seedlings compared to 2% in the control). Levels of disease suppression increased with the incubation time prior to planting. Disease control effect of CDS was not consistent between different batches of peat-based mix. In a non-suppressive batch of peat-based mix, disease suppression by CDS was enhanced by a bio-control agent, Trichoderma hamatum 382. In muck soil from a commercial field naturally-infested with Pythium spp., CDS (0.25, 0.5, and 1%) provided protection of cucumber seedlings from damping-off immediately after incorporation, but the maximum protection was seen after 1 week with all three rates. The number of total bacteria was enhanced in the CDS-amended muck soil. In the micro-plots, CDS (0.5 and 1%) as an amendment to muck soil 2 weeks before planting improved the percentage of healthy cucumber seedlings and fresh plant weight compared to the control. CDS is not toxic to the pathogens and disease suppression is believed to be due to biological activity stimulated by CDS in the substrate.     

Species of Phytophthora and Pythium as Nematode-destroying Fungi

Pythium monospermum,, P. aphanidermatum, and Phytophthora palmivora were found to be capable of destroying certain nonstylet-bearing nematodes through endozoic parasitism by hyphae from ingested zoospores. Hyphae of P. monospermum parasitized nematode eggs but could not capture or otherwise prey upon living nematodes. We suggest that endoparasitism of free-living nematodes may be common among Oomycetes in nature.