Antagonistic Pythium against pathogenic Pythium on cucumber roots1

Antagonistic microorganisms introduced into the growing medium, the seed or the root, have been shown to reduce the attack of many soil-borne plant pathogenic fungi. In 1983, three experiments with Pythium oligandrum were carried out using cucumber seedlings artificially inoculated with Pythium splendens. The seedlings cv. Ideal Nova were grown in shallow boxes containing granulated rockwool fibre (Grodania). The plants were grown at a minimum temperature of 20°C in a glasshouse. Plants were treated with the antagonist either by coating the seeds, or by applying a disc of a PDA-culture to 9-day old seedlings. The pathogen was introduced 9 or 5 days later, respectively, by adding PDA culture discs to the rockwool; 15 and 38 days later, the area with attacked roots was calculated as a percentage of the box-bottom area. The experiments showed that it is possible to control P. splendens by either method of application. P. oligandrum was able to colonize the seed and rhizosphere and suppress P. splendens for at least 38 days. If future experiments are similarly positive, the methods for use in commercial horticulture should be worked out as soon as possible.     

Effect of salinity on pythium damping-off of cucumber and on the tolerance of Pythium aphanidermatum

A study was conducted to investigate the effect of salinity on pythium damping-off and tolerance to salinity among Pythium aphanidermatum isolates from different geographical origins, genetic backgrounds and saline/non-saline soils. Increasing irrigation-water salinity from 0·01 to 5 dS m⁻¹ significantly increased mortality in cucumber seedlings inoculated with P. aphanidermatum and reduced dry weight of non-inoculated seedlings. In vitro tests in culture media amended with different concentrations of NaCl showed that growth of P. aphanidermatum, P. spinosum and P. splendens isolates was stimulated or unaffected at salinity levels stressful for cucumber (electrical conductivity = 5 dS m⁻¹). Significant differences were found in tolerance to salinity among 47 P. aphanidermatum isolates obtained from different greenhouses. The concentration of NaCl required to reduce growth of P. aphanidermatum isolates by 50% varied from 23 to 62 dS m⁻¹, with an average of 46 dS m⁻¹, however, oospore production was more sensitive to salinity and no oospores were produced above 20 dS m⁻¹. Analysis of tolerance of P. aphanidermatum isolates to salinity showed no relationship between tolerance to salinity and geographical origins or genetic data. Isolates of P. aphanidermatum from greenhouses with no salinity problems were as tolerant to salinity as isolates obtained from salinity-affected greenhouses, suggesting lack of evidence for ecological adaptation within P. aphanidermatum in Oman. Increased mortality in cucumber seedlings at higher salinity levels may imply a synergistic interaction between salinity stress and salinity-tolerant Pythium species on cucumber seedlings, resulting in greater seedling losses.     

Control of root zone pH is not effective in preventing Pythium aphanidermatum disease in cucumber

Journal of Plant Diseases and Protection - Pythium aphanidermatum is the most wide-spread root pathogen in greenhouse cucumber. Although environmental condi-tions are known to affect the disease...     

Effectiveness of Trichoderma spp. obtained from re-used soilless substrates against Pythium ultimum on cucumber seedlings

Journal of Plant Diseases and Protection - Thirty-nine Trichoderma strains isolated from effective re-used substrates used in soilless systems and two commercial formulations (Trichoderma viride...     

Suppressive effect of composts from residual biomass on Pythium ultimum

Journal of Plant Diseases and Protection - Two experiments were established in the laboratory of the Department of Organic Farming and Cropping Systems and the greenhouse of the University of...     

Evaluation of a method for quantification of Pythium aphanidermatum in cucumber roots at different temperatures and inoculum densities

Journal of Plant Diseases and Protection - A method for the quantification of Pythium aphanidermatum density in cucumber roots based on an indirect enzyme-linked immunosorbent assay (Elisa) was...     

Molecular characterization and pathogenicity of Pythium species associated with damping-off in greenhouse cucumber (Cucumis sativus) in Oman

A study was undertaken in 2004 and 2005 to characterize pathogens associated with damping-off of greenhouse-grown cucumber seedlings in 13 districts in Oman. Identification of Pythium to the species level was based on sequences of the internal transcribed spacer (ITS) of the ribosomal DNA. Of the 98 Pythium isolates collected during the survey, Pythium aphanidermatum , P. spinosum , P. splendens and P. oligandrum accounted for 76%, 22%, 1% and 1%, respectively. Pythium aphanidermatum was isolated from all of the districts, while P. spinosum was isolated from seven districts. Pathogenicity tests showed inter- and intraspecific variation in aggressiveness between Pythium species. Pythium aphanidermatum , P. spinosum and P. splendens were found to be highly aggressive at 25°C. However, the aggressiveness of P. spinosum decreased when the temperature was raised to 30°C, which was found to correspond to the lower frequency of isolation of P. spinosum in the warmer seasons, compared to the cooler time of the year. Pythium aphanidermatum exhibited limited intraspecific variation in the sequences of the ITS region of the rDNA and showed 100% similarity to the corresponding P. aphanidermatum sequences from GenBank. The ITS sequence data, as well as morphological characteristics of P. spinosum isolates, showed a high level of similarity within and between P. spinosum and P. kunmingense , and suggested that the two species were synonymous. This study represents the first report of P. spinosum, P. splendens and P. oligandrum in Oman.     

Protection of cucumber against Pythium root rot by fluorescent pseudomonads: predominant role of induced resistance over siderophores and antibiosis

Four Pseudomonas strains were evaluated for their intrinsic properties conferring their ability to protect long English cucumber against Pythium aphanidermatum in hydroponic culture. Two of the strains, BTP1 and its siderophore-negative mutant M3, increased plant yield as compared with the non-inoculated control plants. Strain BTP7 was intermediate in its biocontrol activity while strain ATCC 17400 failed to reduce disease development. The role of pyoverdines could not be confirmed since treatment with either BTP1 or its siderophore-negative mutant M3 provided similar suppression of Pythium disease. In addition, no siderophores were detected in the nutrient solution. BTP1 did not inhibit pathogen growth in vitro on several media, suggesting that antibiosis was not a mechanism of suppression. Quantification of root bacterial populations did not indicate differences among the strains. On the other hand, roots treated with either BTP1 or its sid⁻ mutant M3 contained more antifungal phenolics than roots from any other treatments including controls. These results suggest that antifungal compounds induced by inoculation of cucumber roots with the fluorescent Pseudomonas strains BTP1 and M3 participate actively in the protection of cucumber plants against P. aphanidermatum     

Production of tobacco mosaic virus in and its infectivity from leaves of two Nicotiana species treated with 6-azauracil

A correlation between the amount of virus as determined by infectivity test and that determined by a serological-spectrophotometric method was established in leaf discs ofNicotiana tabacum White Burley which had floated on a solution of 6-azauracil (AzU) before and after inoculation with tobacco mosaic virus. More virus, determined in the two different ways, was present in the AzU-than in the water-treated leaf discs at 24 and 48 h after inoculation. Thereafter, the amount of virus was less than that in the control. The inhibitory effect of the pyrimidine analogue on the amount of infectivity in sap from AzU-treated discs proved to be dependent on the dilution of sap used. The higher the dilution the more inhibition occurred.In inoculated leaf discs ofNicotiana glutinosa the formation of local lesions in the AzU-treated series was strongly inhibited at 48 h after inoculation whereas at that time the amount of virus determined by the serological-spectrophotometric method was higher than that in the water-treated control ones. The latter mentioned stimulating effect disappeared later than 48 h after inoculation and gave way to inhibition.The infectivity of tobacco mosaic virus from AzU-treated leaf discs of White Burley tobacco was 23% of that from water-treated controls when undiluted clarified sap was used.     

Aggressiveness and production of cell-wall degrading enzymes by Pythium violae, Pythium sulcatum and Pythium ultimum, responsible for cavity spot on carrots

This study investigated the relationships between pathogenesis, types of symptoms and in vitro production of cell-wall degrading enzymes by P. violae, P. sulcatum and P. ultimum. The three pathogens, considered as the three Pythium species principally responsible for cavity spot on carrot roots, secreted only low levels of fatty acid esterases activity, suggesting they have limited ability to degrade suberin in the walls of the outer cell layers of carrot tissues. Among the enzymes that degrade cell-wall polysaccharides, only pectate lyases and cellulases were produced by P. violae, and these were produced late and in small amounts: the symptoms caused by P. violae were limited and typical of cavity spot. Conversely, P. ultimum caused maceration of tissues, and secreted polygalacturonases and -1,4-glucanases earlier and in larger amounts than P. violae. P. ultimum also produced a large diversity of proteins and cellulase isoenzymes. Although secreting all the monitored enzymes in higher quantity than the two previous species, P. sulcatum was responsible for only typical limited symptoms of cavity spot, with a brown colouring. The role of plant reactions induced in response to early pectinolytic enzyme production by P. sulcatum may account for this apparent inconsistency.Died on April 11, 1997     

Species of Pythium in the Netherlands

A review is given on observations ofPythium spp. so far published. In personal investigations about 4000 isolates ofPythium species were obtained from soil, water, Daphnias and plants during 1966–1972. Special attention was paid to the progressive colonization of the newly reclaimed polder Zuidelijk Flevoland: the numbers of species per sample increased from 0 in 1966 to 13 in 1972. The percentage of samples which did not contain anyPythium decreased from 54 in 1968 to 0 in 1972. From Zuidelijk Flevoland altogether 21 species were isolated, from Oostelijk Flevoland 22, of which 19 occurred in both the polders. The soil depth down to 10 cm had little influence on the occurrence ofPythium species. From soils near Rhenen and Zutphen 12 and 19 species respectively were isolated, from flax and flaxfields 11, from forests 8, from roots of different plants 17 and from water 11. Besides species already previously recorded, 15 new records ofPythium species for the Netherlands are documented. Species with filamentous non-swollen sporangia preferentially occurred in water and wet soils while species with spherical sporangia or hyphal swellings dominated in cultivated soils, forests and in the roots of phanerogams. In 1968 species with filamentous non-swollen sporangia comprised 89% of the isolations from Zuidelijk Flevoland; this percentage decreased to 9 in 1972. The percentage of isolates with spherical sporangia or hyphal swellings from cultivated soils, forest soils or plant material was mostly over 90.P. rostratum Butler, though found in all kinds of soils, was often the only species which could be isolated from dry sandy soil samples.P. sylvaticum Campbell & Hendrix was the most common species in cultivated soils in the Netherlands (37% of the isolates in 1968–1972), followed byP. oligandrum Drechsler (22%) andP. paroecandrum Drechsler (11.5%).     

Association of Pythium coloratum and Pythium sulcatum with cavity spot disease of carrots in Western Australia

All 170 Pythium isolates from carrot cavity spot lesions from a field in Western Australia were found to belong to either P. coloratum or P. sulcatum . All isolates of P. coloratum produced large, brownish-black, water-soaked and depressed lesions on mature carrots inoculated with agar plugs colonized by the pathogen. In comparison, only a few isolates of P. sulcatum produced lesions and these were small. In glasshouse trials, P. coloratum produced substantial and numerous lesions at an inoculum density of 0.5% (weight of millet seed-based inoculum/weight of soil), whilst P. sulcatum produced few and small lesions at inoculum densities of 0.8 and 1% and none at 0.5%. This is the first record of P. coloratum as a causal agent of cavity spot of carrots.     

A microscopic study of the wheat-powdery mildew relationship after application of the systemic compounds procaine,griseofulvin and 6-azauracil

On wheat seedlings systemically protected by root application of procaine-hydrochloride, griseofulvin or 6-azauracil, germination of oidia ofErysiphe graminis f. sp.tritici and penetration of this fungus into the epidermal cell wall was as high as on control plants. Inhibition of powdery mildew development became apparent only after the penetration process had started. About 85% of the infections were halted within 24 hours after the inoculation, and did not result in the formation of a haustorium. In the other cases usually not more than one haustorium per infection court was formed, which often showed anomalies, characteristic for each compound used. Development of mycelium was scanty or absent and no sporulation occurred. Similarly, on plants of a resistant wheat variety, powdery mildew inhibition became apparent only after penetration of the host had started. There was no development of mycelium or sporulation. A severe reaction of certain epidermal cells to penetration by powdery mildew was observed on resistant as well as on treated and untreated susceptible plants. However, in relation to the total number of infections, the percentage of this type of reaction was low.     

Factors affecting the control of Pythium ultimum damping-off of sugar beet by Pythium oligandrum

Application of Pythium oligandrum to a soil-based compost as a mycelial suspension (5 × 10² CFU g⁻¹ of dry compost) and oospore alginate pellets (10⁵ oospores/g of dry compost) controlled pre- and postemergence damping-off of sugar beet caused by Pythium ultimum to a level similar to metalaxyl seed treatment. Oospore seed treatments and aqueous suspensions of oospores applied to compost failed to control disease. Problems in the use of P. oligandrum oospore inocula for the control of damping-off were highlighted. It was shown that treatment of oospores with cellulase (20 g L⁻¹) increased germination approximately three-fold in comparison to untreated spores. Untreated and cellulase pretreated oospores were subsequently evaluated as seed treatments for their ability to control damping-off of sugar beet. The highest rate of pretreated oospores (10⁴ oospores/seed) gave levels of emergence and establishment in infested compost that were not significantly different from the uninfested controls, whereas seed treatment with untreated oospores gave no significant reduction in disease. In a trial carried out in a controlled environment to assess the effect of pH (4.5–8.0), P. oligandrum (10⁴ cellulase pretreated oospores/seed) was shown to control pre- and postemergence damping-off of sugar beet at pH 7.0 and 7.5 only.     

Inoculation of cucumber roots with zoospores of mycoparasitic and plant pathogenic Pythium species: Differential zoospore accumulation, colonization ability and plant growth response

Hydroponically grown cucumber (Cucumis sativus) seedlings were inoculated with zoospores of 1 mycoparasitic (Pythium oligandrum) and 2 pathogenic (Pythium aphanidermatum and Pythium group F) Pythium spp. During the first 2 days after inoculation, all the Pythium spp. caused reduction in the root length. However, roots treated with Pythium oligandrum quickly reached the length of the control and on the 8th day, and for the rest of the experimental period, stimulation of root elongation was noted. Pythium oligandrum was not pathogenic on cucumber and no differences in the fresh weights of control and Pythium oligandrum inoculated plants were observed in the course of the experiment. Pythium group F and Pythium aphanidermatum were pathogenic on cucumber seedlings, but their pathogenicities differed. Thus, while Pythium group F had a constant, negative influence on root length and plant growth, measured as fresh weight, Pythium aphanidermatum caused generalized necroses of the root system, inhibiting consistently root elongation and plant growth and finally causing plant death. Moreover, the zoospores of 2 mycoparasitic species, Pythium oligandrum and Pythium periplocum, were not attracted to roots of cucumber and accumulated on the roots in very low numbers compared to those of the pathogenic species, Pythium aphanidermatum, which were strongly attracted and accumulated in large numbers. Finally, it was also found that Pythium oligandrum colonized the roots very poorly, while Pythium group F and Pythium aphanidermatum were significantly better root colonizers. The significance of these findings is discussed in relation to the ecology of Pythium species and biocontrol.     

Fungicide failure to control Pythium blight on turf grass in Italy

Journal of Plant Diseases and Protection - Pythium blight is one of the most important turf grass diseases. During the past three years, the failure of fungicide treatments against the disease has...     

Pythium violae isolated from cavity spot lesions on carrots in Israel

Pythium violae Chesters & Hickman was isolated for the first time from cavity spot lesions on carrots in Israel and the resulting isolates were compared with isolates from the UK, France and Spain. The isolates from all locations responded similarly to temperature and had similar colony morphology, oogonial diameters and soluble protein patterns.