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PubMed2005

[Population fluctuation of main pathogens and their antagonistic bacteria in cucumber rhizosphere].

The investigation on the population fluctuation of main pathogens Pythium spp. and Fusarium oxysporum f. cucumerinum in the rhizosphere of four cucumber varieties showed that there was a significant difference in population number of pathogens for different cucumber varieties and for different growth stages of the same variety, which was in accordance with the regular disease incidence pattern. The population number of Pythium spp. was significantly higher at cucumber creeping stage than at its other growth stages, while that of Fusarium oxysporum f. sp. cucumerinum was the highest at cucumber principal-fruiting stage. There was a closer positive relationship between two test pathogens and their antagonistic bacteria, with the correlation coefficient being 0.95 and 0.81, respectively.

Antibiosis↗

PubMed2005

Armillaria mellea as a cause of oak decline in Hatam-baigh forest of Iran.

A200 ha forest of "Hatam-baig" is located in Ardebil Province on the Northwest of Iran. Oak trees (Quercus macranthera Fisch & Mey) in this forest have been faced with declining and extinction since 1991, that has destructed about one third of the forest trees until now. This disorder was expressed in various symptoms including wilting, defoliation and decline. In order to identify factors causing decline, a study was managed from 1998 to 2001. Samples were taken from roots, trunks, crowns and soil beneath the canopy and were cultured on different culture media subsequently. Armillaria mellea (Vahl) P. Kumm., Phytophthora cryptogea Pethybr. & Laff., Dematophora sp., Pythium aphanidermatum (Edson) Fitzp. and Fusarium spp. were the most common isolated fungi. A. mellea appeared to be the essential causal agent of the decline according to the studies made on oak tress decline around the world and based on brown rot observed beneath mycelial fans in the cross section prepared from the trunk and characteristics of the isolated fungi. The fungus activity had been favored by physiological weakness and stresses in oak rootstocks caused by brown- tail moth (Euproctis chrysorhoea L.) and drought stress in infected trees. The biological species of this fungus was identified as Armillaria mellea, using hybridization tests and application of haploid test strains. The fungi such as Phytophthora sp., Pythium sp., and Dematophora sp. can not be infective in this forest due to being hydrophylous. In the southern part of the forest with remarked steepness, the severity of the decline appears to be more than that in the smoothly northern part. The decline of Q. macranthera is reported as matrix nova. The report of the isolated fungi from this oak species is also universally new.

Basidiomycota↗

PubMed1980

Carbon and nitrogen nutrition of plant pathogenic fungi associated with basal stem rots of cowpeas, Vigna unguiculata (L) Walp in Nigeria.

Pythium aphanidermatum, Rhizoctonia bataticola (syn. = Macrophomina phaseolina), Botryodiplodia theobromae, and two strains of Sclerotium rolfsii readily utilized for growth the monosaccharides glucose, fructose and mannose, the disaccharides sucrose and maltose and the polysaccharides dextrin and starch. In addition, S. rolfsii grew appreciably well on carboxymethylcellulose (CMC). When filter paper (nature cellulose) was supplied as sole carbon source, it was decomposed by B. theobromae, S. rolfsii (SR2) and R. bataticola. In studies on nitrogen utilization, B. theobromae grew best on glutamic acid, P. aphanidermatum on glycine and the two strains of S. rolfsii on ammonium nitrate. R. bataticola utilized casein hydrolysate most effectively for growth. EDTANa2 was a poor nitrogen source for all the pathogens.

Amino Acids↗

PubMed2005

Evaluation of fungicides for the control of carrot cavity spot.

Cavity spot is one of the most common and serious diseases of carrot (Daucus carota L). The disease, caused by different species of Pythium, including P. violae Chesters & Hickman, P. sulcatum Pratt & Mitchell and P. sylvaticum Campbell & Hendrix, leads to frequent high rejection rates during grading worldwide. In the area of the city of Québec, the disease is caused mainly by P. sulcatum, P. sylvaticum and P. macrosporum Vaartaja & van der Plaats-Niterink. Cavity spot can be controlled with metalaxyl, but reports are emerging that this treatment show little or no efficacy in many regions. This situation reinforces the need for alternative fungicides. The objectives of the present study were: (1) to determine the sensitivity of 14 pathogenic isolates of P. sulcatum, P. sylvaticum and P. macrosporum collected from carrots produced in the area of the city of Québec to different broad-spectrum and oomycete-specific fungicides (chlorothalonil, etridiazole, fludioxonil, fosetyl-Al, metalaxyl, zoxamide), (2) to evaluate the efficacy of the fungicides in controlling cavity spot, and (3) to evaluate the risk of resistance development of isolates with the best-performing fungicide(s). The determination of EC50 for the fungicides tested showed that most isolates were highly sensitive to both metalaxyl and zoxamide but insensitive to fludioxonil, fosetyl-Al and chlorothalonil. In greenhouse assays, only zoxamide provided significant and consistent disease control as measured by the number of cavity spot lesions caused by P. sulcatum. Investigations into the risk of resistance development to zoxamide showed that, for specific isolates, repeated exposure to the fungicide resulted in a loss of sensitivity.

Daucus carota↗

PubMed2004

Synthesis and fungicidal activity of ethaboxam against Oomycetes.

This study describes the chemical synthesis and intrinsic fungicidal activity of ethaboxam [(RS)-N-(alpha-cyano-2-thenyl)-4-ethyl-2-(ethylamino)-1,3-thiazole-5-carboxamide], a new Oomycetes fungicide. In in vitro tests, ethaboxam showed inhibitory activity against isolates of Phytophthora and some Pythium spp, with MIC values ranging from 0.1 to 0.5 mg litre(-1) for nine isolates of Phytophthora infestans (Montagne) de Bary and from 1.0 to 5.0 mg litre(-1) for eight isolates of Phytophthora capsici Leonian. In tests to determine time and concentration for complete inactivation of each pathogen (five isolates of P infestans and five isolates of P capsici), ethaboxam inactivated all isolates of P infestans within 48h at 10 mg litre(-1) and those of P capsici within 96 h at 10 mg litre(-1). Ethaboxam effectively suppressed development of tomato late blight caused by P infestans and pepper Phytophthora blight caused by P capsici in the studies conducted to determine its preventive, curative, persistent and systemic activity. These results show that ethaboxam has desirable fungicidal characteristics as an Oomycetes fungicide.

Fungicides, Industrial↗

PubMed1990

Antagonistic potential of Gliocladium virens and Trichoderma longibrachiatum to phytopathogenic fungi.

Three isolates of Gliocladium virens (G1, G2 and G3) and two of Trichoderma longibrachiatum (T1 and T2) were screened against isolates of three soilborne plant pathogens namely Rhizoctonia solani, Sclerotium rolfsii and Pythium aphanidermatum. G. virens exhibited stronger hyperparasitism and wider biological spectrum than T. longibrachiatum. Further, similarities as well as variation was observed in the ability of the various isolates to invade the test pathogens in dual culture. For the hyperparasites, acidic pH range (5.0 to 5.5) favoured both growth and spore germination. The hyperparasites made direct contact with the pathogens followed by varied modes of attack invariably leading to cell disruption. Antagonists, G1 and G3 revealed strong antibiosis while T2 showed moderate effect. All the isolates produced enhanced levels of lytic enzymes adaptively and there were marked differences among them. However, no correlation was observed between these attributes and the hyperparasitic potential of the various isolates in dual culture. The relevance and the role of enzymes and toxic metabolite(s) in the antagonism of G. virens and T. longibrachiatum to these pathogens are discussed.

Chytridiomycota↗

PubMed1986

Equine pythiosis in Costa Rica: report of 39 cases.

Thirty-nine pythiosis equine cases, were studied at the Veterinary Medicine School of the National University of Costa Rica, between 1981 and 1984. Lesions were located in different parts of their anatomy: anterior and posterior extremities, abdomen, thorax, breast and mammary gland, and were characterized by their tumoral appearance with necrotic tissue in which yellow-white coral-like necrotic masses, called kunker or leeches were shown. Splendore-Hoeppli like phenomenon and eosinophilic inflammatory reaction around the hyphae, was microscopically observed. Pythium sp. (Hyphomyces destruens) was isolated in Sabouraud dextrose agar from ground kunkers. Immunodiffusion (ID) to diagnose this disease in equines, was performed with success. Immunotherapy was applied to 5 of affected horses, and three were cured. Some epizootiological aspects of the pythiosis are also discussed.

Animals↗

PubMed1992

Evaluation of two vaccines for the treatment of pythiosis insidiosi in horses.

Two vaccines to treat phythiosis insidiosi in horses were evaluated in 71 Costa Rican horses between 1982 to 1988. One vaccine used a cell-mass (CMV) as antigen and the other a soluble concentrated antigen (SCAV). Both vaccines cured horses infected with Pythium insidiosum (p value approximately 14%). The age of lesions prior to vaccination was important in the response of the horses to immunotherapy. All horses with lesions 0.5 months or less in duration were cured regardless of the vaccine used. Horses with lesions two or more months old did not respond to either vaccine. The age of the horses did not have any influence on their response to the vaccinations. The CMV produced a prominent inflammatory reaction at the side of injection, while the SCAV gave a low inflammatory reaction. In addition, the CMV lost its effectiveness two to three weeks after its preparation. By contrast, the SCAV maintained its ability to cure horses even after 18 months. Immunotherapy using SCAV can thus be used as the vaccine of choice in early cases of equine cutaneous pythiosis insidiosi.

Aging↗

PubMed1995

Fungal diseases of fish in Nanak Sagar, Naini Tal, India.

Eight zoosporic fungi viz., Achlya debaryana, A. flagellata, A. klebsiana, Aphanomyces laevis, Saprolegnia diclina, S. ferax, S. parasitica, and Pythium sp. were isolated from a large number of adult fishes of the species Mastacembelus armatus, Mystus vitatus, Nandus nandus, Tor putitora and T. tor of Nanak Sagar reservoir in Naini Tal district, India. Species of the parasites and the hosts were different in their pathogenicity and immunity, respectively. However, A. flagellata and S. parasitica appeared to be the most virulent. The severity of mycosis was primarily correlated to moderate water temperatures of 22-25 degrees C. High temperature (> 28 degrees C) retarded the disease process. The experimental inoculation with all the associated fungal species on Puntius conchonius in the laboratory produced clinical signs similar to the ones seen on infected fish in the reservoir. This is the first report on fish mycosis in the large reservoir located in the foot hill of Kumaun Himalaya.

Animals↗

PubMed1995

Experimental transmission and pathogenicity of some zoosporic fungi to Tilapia fish.

Seventeen fungal species belong to sex genera were recovered from the four organs of Tilapia fish and the most common were Saprolegnia ferax, S. diclina, Achlya dubia, A. americana, A. racemosa and A. flagellata, Dictyuchus sterile, Pythium undulatum and Aphanomyces sp. Severe infection followed by death of all fish was incited by S. parasitica and S. ferax through experiment I. 30-70% of T. nilotica and T. galileae were killed through experiment II by S. parasitica and S. ferax. T. galileae was more susceptible to fungal infection than T. nilotica.

Animals↗

PubMed2006

Rhizosphere Pseudomonas sp. strains reduce occurrence of pre- and post-emergence damping-off in chile and tomato in Central Himalayan region.

Based on in vitro screening for PGP and anti-mycelial activity against three zoosporic pathogenic oomycetes, Pythium aphanidermatum 123, P. aphanidermatum 4746, and Phytophthora nicotianae 4747, seven bacterial isolates were selected for field trials on tomato and chile to test for plant growth promotion under natural and artificial disease-infested field sites in both winter and wet seasons. The effectiveness of isolates in the field trials correlated with the in vitro antagonism screening data. Pseudomonas sp. FQP PB-3, FQA PB-3 and GRP(3) showed substantial beneficial effects on plant growth promotion and lowered considerably the incidence of pre- and post-emergence damping-off in both the crops under various disease scenarios. For example, seed bacterization with these bacterial strains reduced pre-emergence-damping off by ca. 60-70% in the two natural sites, with and without histories of fungicide use in the winter season, and to a lesser extent, ca. 20-40%, in the warmer wet (high humidity; 85-92%) season. The suppression efficacy for post-emergence damping-off was less compared to pre-emergence damping-off although still significant (P > 0.05). Our data unambiguously show that screening of a large number of bacterial pool identifies promising isolates that show beneficial effects on all stages of plant growth in natural oomycete-infested regimes.

Antibiosis↗

PubMed2004

Bacillus subtilis M4 decreases plant susceptibility towards fungal pathogens by increasing host resistance associated with differential gene expression.

Results presented in this paper describe the ability of Bacillus subtilis strain M4 to reduce disease incidence caused by Colletotrichum lagenarium and Pythium aphanidermatum on cucumber and tomato, respectively. Disease protection in both pathosystems was most probably due to induction of resistance in the host plant since experiments were designed in order to avoid any direct contact between the biocontrol agent and the pathogen. Pre-inoculation with strain M4 thus sensitised both plants to react more efficiently to subsequent pathogen infection. In cucumber, the use of endospores provided a disease control level similar to that obtained with vegetative cells. In contrast, a mixture of lipopeptides from the surfactin, iturin and fengycin families showed no resistance-inducing potential. Interestingly, treatment with strain M4 was also associated with significant changes in gene transcription in the host plant as revealed by cDNA-AFLP analyses. Several AFLP fragments corresponded to genes not expressed in control plants and specifically induced by the Bacillus treatment. In support to the macroscopic protective effect, this differential accumulation of mRNA also illustrates the plant reaction following perception of strain M4, and constitutes one of the very first examples of defence-associated modifications at the transcriptional level elicited by a non-pathogenic bacterium in a host plant.

Bacillus subtilis↗

PubMed2005

Involvement of fengycin-type lipopeptides in the multifaceted biocontrol potential of Bacillus subtilis.

In this work, the potential of Bacillus subtilis strain M4 at protecting plants against fungal diseases was demonstrated in different pathosystems. We provide evidence for the role of secreted lipopeptides, and more particularly of fengycins, in the protective effect afforded by the strain against damping-off of bean seedlings caused by Pythium ultimum and against gray mold of apple in post-harvest disease. This role was demonstrated by the strong biocontrol activity of lipopeptide-enriched extracts and through the detection of inhibitory quantities of fengycins in infected tissues. Beside such a direct antagonism of the pathogen, we show that root pre-inoculation with M4 enabled the host plant to react more efficiently to subsequent pathogen infection on leaves. Fengycins could also be involved in this systemic resistance-eliciting effect of strain M4, as these molecules may induce the synthesis of plant phenolics involved in or derived from the defense-related phenylpropanoid metabolism. Much remains to be discovered about the mechanisms by which Bacillus spp suppress disease. Through this study on strain M4, we reinforce the interest in B. subtilis as a pathogen antagonist and plant defense-inducing agent. The secretion of cyclic fengycin-type lipopeptides may be tightly related to the expression of these two biocontrol traits.

Antibiosis↗

PubMed2003

An antifungal chitinase produced by Bacillus cereus with shrimp and crab shell powder as a carbon source.

The production of inexpensive chitinolytic enzymes is an element in the utilization of shellfish processing wastes. In this study, shrimp and crab shell powder prepared by treating shrimp and crab processing wastes with boiling and crushing was used as a substrate for the isolation of an antifungal chitinase-producing microorganism. Bacillus cereus YQ 308, a strain isolated from the soil samples, excreted one chitinase when cultured in a medium containing 2% (wt/vol) shrimp and crab shell powder as major carbon source. The chitinase, purified by sequential chromatography, had an Mr of 48 kDa and pI of 5.2. The purified chitinase (2 mg/ml) inhibited the hyphal extension of the fungi Fusarium oxysporum and Pythium ultimum.

Animals↗

PubMed1998

Regulation of enzymes involved in anthocyanin biosynthesis in carrot cell cultures in response to treatment with ultraviolet light and fungal elicitors.

The accumulation of anthocyanins in cell cultures of Daucus carota L. and the enzymes involved in their biosynthesis were investigated under growth in the dark, continuous irradiation with UV light, incubation with elicitors from Pythium aphanidermatum, and elicitor treatment of UV-irradiated cells. Upon UV irradiation, anthocyanin accumulation was strongly enhanced, and the enzymes of the phenylpropanoid and flavonoid pathways, including the "late" enzymes cyanidin galactosyltransferase, cyanidin galactoside xylosyltransferase, cyanidin triglycoside sinapoyltransferase and sinapic acid glucosyltransferase, all showed transient increases in their activities. The time courses of the enzyme activities exhibited successive maxima with an ordered sequence corresponding to their position in the biosynthetic pathway, suggesting a coordinated induction of the entire set of enzymes. The key enzymes phenylalanine ammonia-lyase and chalcone synthase are regulated on a transcriptional level. Incubation of dark-grown carrot cells with fungal elicitors led to a rapid and transient induction of phenylalanine ammonia-lyase corresponding to the formation of 4-hydroxybenzoic acid, but the amount of anthocyanin did not increase and there was no enhancement of any of the enzyme activities which are part of the anthocyanin pathway, including the enzymes catalyzing glycosylation and acylation reactions. Treatment with UV light and elicitors resulted in a rapid induction of the phenylpropanoid pathway, whereas the inducing effect of UV light on the anthocyanin content, on chalcone synthase and on the enzymes catalyzing the final steps of anthocyanin biosynthesis was suppressed. These results indicate a coordinated regulation of the enzymes involved in anthocyanin biosynthesis, an independent inducibility of the phenylpropanoid pathway, and a hierarchy of the different effectors, as shown by the dominating role of the elicitor-signal over the UV stimulus.

Anthocyanins↗

PubMed2005

Characterization and mapping of Rpi1, a gene that confers dominant resistance to stalk rot in maize.

The maize inbred lines 1145 (resistant) and Y331 (susceptible), and the F(1), F(2) and BC(1)F(1) populations derived from them were inoculated with the pathogen Pythium inflatum Matthews, which causes stalk rot in Zea mays. Field data revealed that the ratio of resistant to susceptible plants was 3:1 in the F(2) population, and 1:1 in the BC(1)F(1)population, indicating that the resistance to P. inflatum Matthews was controlled by a single dominant gene in the 1145xY331 cross. The gene that confers resistance to P. inflatum Matthews was designated Rpi1 for resistance to P. inflatum) according to the standard nomenclature for plant disease resistance genes. Fifty SSR markers from 10 chromosomes were first screened in the F(2) population to find markers linked to the Rpi1 gene. The results indicated that umc1702 and mmc0371 were both linked to Rpi1, placing the resistance gene on chromosome 4. RAPD (randomly amplified polymorphic DNA) markers were then tested in the F(2)population using bulked segregant analysis (BSA). Four RAPD products were found to show linkage to the Rpi1 gene. Then 27 SSR markers and 8 RFLP markers in the region encompassing Rpi1 were used for fine-scale mapping of the resistance gene. Two SSR markers and four RFLP markers were linked to the Rpi1 gene. Finally, the Rpi1 gene was mapped between the SSR markers bnlg1937 and agrr286 on chromosome 4, 1.6 cM away from the former and 4.1 cM distant from the latter. This is the first time that a dominant gene for resistance to maize stalk rot caused by P. inflatum Matthews has been mapped with molecular marker techniques.

Chromosome Mapping↗

PubMed2005

Sodium, potassium-atpases in algae and oomycetes.

We have investigated the presence of K(+)-transporting ATPases that belong to the phylogenetic group of animal Na(+),K(+)-ATPases in the Pythium aphanidermatum Stramenopile oomycete, the Porphyra yezoensis red alga, and the Udotea petiolata green alga, by molecular cloning and expression in heterologous systems. PCR amplification and search in EST databases allowed one gene to be identified in each species that could encode ATPases of this type. Phylogenetic analysis of the sequences of these ATPases revealed that they cluster with ATPases of animal origin, and that the algal ATPases are closer to animal ATPases than the oomycete ATPase is. The P. yezoensis and P. aphanidermatum ATPases were functionally expressed in Saccharomyces cerevisiae and Escherichia coli alkali cation transport mutants. The aforementioned cloning and complementary searches in silicio for H(+)- and Na(+),K(+)-ATPases revealed a great diversity of strategies for plasma membrane energization in eukaryotic cells different from typical animal, plant, and fungal cells.

Amino Acid Sequence↗

PubMed2005

Intestinal canine pythiosis in Venezuela confirmed by serological and sequencing analysis.

An 11-months-old mixed Terrier male originally from Venezuela, was referred to a Veterinary Hospital with signs of depression, anorexia, vomiting and diarrhea. The illness had begun 1 month earlier. Despite antibiotic chemotherapy and vitamins, the disease progressed. Radiological exams showed involvement of the small intestine. Histopathological studies of tissue samples taken during surgical intervention revealed eosinophilic areas in the center of which, abundant eosinophils, histiocytes and giant cells were observed. Silver stained cross-sections of the small intestine showed slender sparsely septate hyphae within the necrotic areas. Attempts to isolate the etiologic agent in pure culture were fruitless. The dog died without a definitive diagnosis. Fixed tissue samples of the small intestine were later investigated using specific fluorescent antibodies for pythiosis and molecular tools. These exams indicated that the hyphae in the infected tissues belong to the straminipilan pathogen Pythium insidiosum. This is the first confirmed case of dog pythiosis in Venezuela.

Animals↗