Selection of bacterial antagonists for the biological control of Botrytis cinerea in apple (Malus domestica) and in comparison with application of thiabendazole.

In this study fifteen strains of identified Pseudomonas fluorescens and Bacillus subtilis were investigated for biological control activity against Botrytis cinerea. P. fluorescens P-35 and B. subtilis B-16 showed the most inhibitory zone in dual culture assay against B. cinerea, In vitro. After ten days, P. fluorescens P-5 and B. subtilis B-3 showed the considerable results against B. cinerea on apple fruits and could reduce the grey mould from 100% to less than 35%. After twenty days, P. fluorescens P-5 and 8. subtilis B-16 decreased the disease from 100% to less than 65%. Also, application of thiabendazol at 1500 mg/litre was more effective and could reduce the disease from 100% to 30% and 60%, after 10 and 20 days respectively. Results indicated that there is no significant difference among the treatments (thiabendazol and bacterial strains). So, bacterial strains could not only control the disease but also be a reliable replacement instead of Thiabendazol.

Screening of Pseudomonas and Bacillus isolates for potential biocontrol of the damping-off of bean (Phaseolus coccineus).

In this study fifteen isolates of identified Pseudomonas fluorescens and Bacillus subtilis were investigated for control of bean damping-off disease caused by Rhizoctonia solani. In vitro, P. fluorescens P-6 and B. subtilis B-3 showed the most inhibitory zone in dual culture assay against R. solani. The growth of P-6 (4.5 x 10(8) cfu/ml) was significantly higher than in the other treatments. In greenhouse condition, all of the isolates effectively controlled damping-off on bean. P. fluorescens P-5 and P-6 showed the considerable results against R. solani and could reduce the damping-off disease from 100% to less than 30%. P-5, P-6 (P. fluorescens) and 8-16 (8. subtilis) strains had the highest effect on fresh weight of bean.

Effect of essential oils in control of plant diseases.

In this study, antifungal activity of some essential oils, extracted from Syzygium aromoticum, Foeniculum vulgare, Cuminum cyminum and Mentha piperita were investigated against grey mould of apple. The essential oils of S. aromaticum and F. vulgare showed considerable antifungal activities on PDA medium against Botrytis cinerea. Results indicated that the increasing of dosage of the essential oils caused to the more antifungal activity against B. cinerea in vitro condition. After 10 days, results showed that the essential oil of F. vulgare in both of the concentrations (750 and 1000 microL/L) was more effective than the essential oil of S. aromaticum against grey mould of apple and decrease the disease up to 15.5% in comparison with the check treatment (100%). After 20 days, biocontrol potential of the essential oils of S. aromaticum and F. vulgare at 1000 microL/L were more effective than the other treatments and the percentage of disease was evaluated 41.6% and 50.8%, respectively, in comparison with the check treatment (100%).

Interaction of media on production and biocontrol efficacy of Pseudomonas fluorescens and Bacillus subtilis against grey mould of apple.

The medium has a profound effect on biocontrol agents, including ability to grow and effectiveness in disease control. In this study, growth and antagonistic efficacy of strains P-5 and P-35 (P. fluorescens), B-3 and B-16 (B. subtilis) were evaluated in combinations of two carbon (sucrose and molasses) and two nitrogen (urea and yeast extract) sources to optimize control of Botrytis cinerea on apple. All of the strains were grown in different liquid media (pH = 6.9) including: sucrose + yeast extract, molasses of sugar beet + yeast extract in 2:1 and 1:1 w/w ratios, molasses of sugar beet + urea, molasses, malt extract and nutrient broth. Apples (Golden Delicious) were inoculated by a 25-microl suspension of 10(6) spores of B. cinerea per ml, wounding each fruit (in two sites separately). Then a 25-microl suspension of each strain, containing 2 x 10(8) cfu ml(-1) grown in each of the above culture media, was applied to each wound. Results indicated that Molasses + Yeast extract (1:1 w/w) medium supported rapid growth in all of the strains. The final growth of B. subtilis B-16 in Molasses + Yeast extract (1:1 w/w) medium was 5 x 10(9) cfu ml(-1). After ten days, all of the strains significantly inhibited pathogenicity of B. cinerea on apples. The biocontrol efficacy of B. subtilis B-3 in Molasses + Yeast extract (1:1 w/w) medium reduced the severity of grey mould from 100% (inoculated control) to less than 26.9%. After 20 days, Strain B-3 showed a considerable biocontrol efficacy in Molasses medium and reduced the severity of grey mould from 100% (inoculated control) to less than 38.2%. The results obtained in this study could be used to provide a reliable basis for the increase of population of biocontrol agents in fermentation process.

Powder formulation of Burkholderia cepacia for control of rape seed damping-off caused by Rhizoctonia solani.

Talc-based formulation of Burkholderia cepaci strain Bu1 was tested as seed and soil drenchs separately for its ability to control Rhizoctonia soloni the causal agent of rape seed damping-off in greenhouse and field trials. In general, the formulated bacteria was more effective to suppress the disease than the suspension of bacteria cells in carboxymethylcellulose solution (1% w/v), in both greenhouse and field trials. The formulation of strain Bul as soil and seed treatments had the greatest effect on reducing the rape seed damping-off in greenhouse and field trials (66.7, 53.3, 64.4 and 40% respectively). The formulation of strain Bu1 as soil and seed treatments were the most effective treatments to increase the root dry weights in the infected soil in greenhouse. The formulation of strain Bul as soil drench had the greatest effect on enhancement of the fresh weight of roots and stem fresh and dry weights. The formulation of strain Bu1 stored at 4 degrees C exhibited better shelf Life and efficacy in vitro than it's counterpart stored at 25 degrees C.

Evaluation of antifungal activity of carbonate and bicarbonate salts alone or in combination with biocontrol agents in control of citrus green mold.

The aim of this research was to determine if the attacks of green mold on orange could be reduced by edible salts alone or in combination with biocontrol agent. For this purpose toxicity to Pantoea digitatum and practical use of sodium carbonate (SC), sodium bicarbonate (SBC) and potassium carbonate, and potassium bicarbonate alone or in combination with antagonistic bacteria (Pseudomonas fluorescens isolate PN, Bacillus subtilis isolate VHN, Pantoea agglomerans isolate CA) to control green mold were determined. All were fungistatic. SC and SBC were equal and superior to the other salts for control of green mold on oranges inoculated 6h before treatment and were chosen for subsequent trails under cold storage conditions. The biocontrol agents were found completely tolerant to 3% sodium bicarbonate and sodium carbonate at room temperature; although their culturability was reduced by > 1000-fold after 60 min in 1% other salt solutions. Satisfactory results were also obtained with the combined treatment for control of green mold. A significant increase in biocontrol activity of all isolate was observed when combined with sodium carbonate and sodium bicarbonate. The treatments comprising CA combined with SB was as effective as fungicide treatment. Thus, use of sodium bicarbonate treatment at 3% followed by the antagonist P. agglomerans CA could be an alternative to chemical fungicides for control of green mold on oranges.

Evaluation of the effects of chemical versus biological control on Botrytis cinerea agent of gray mould disease of strawberry.

This study investigates on effects of four fungicide and six isolate from Trichoderma and Gliocladium on Botrytis cinerea agent grey mold of strawberry under library and greenhouse condition. The effect of four fungicides i.e. benomyl, dichlofluanid, captan and triadimenol on B. cinerea was studied in the laboratory condition by method mixed poison to culture medium. It was shown that the fungicide including benomyl, triadimenol, dichlofluanid and captan were able to inhibit mycelial growth of B. cinerea on PDA plate with EC50 of 0.16, 1.42, 3.40 and 7.73 ppm respectively. These fungicides delayed myceliogenic germination of sclerotia at 1000 ppm, while exhibiting no fungicidal effect. Moreover, the antagonistic effects of six fungi including Trichoderma koningii (T21), T. viride (T4), T. harzionum (T5), T. viride (T2), G. virens (G2), G. virens (G8) on B. cinerea were assessed. This assessment was done under library condition and its results as follows: The antagonistic mechanism occurred through branching at the end of B. cinerea hyphae, hyphal contact, coiling, vacuolization and lyses. Volatile metabolites of T. koningii (T21) and non-volatile metabolites of G. virens (G2 and G8) and T. koningii (T21) caused maximum inhibition of the fungal growth. Trichoderma spp and G. virens were able to colonize and sporulate on sclerotia and caused their lysis within 7-21 days. In greenhouse, a completely randomized design with 11 treatments (4 chemical and 6 biological and one untreated control) each replicated five times were used for the comparison. Greenhouse studies revealed that application of fungicides i.e. captan, dichlofluanid, triadimenol and benomyl reduces disease severity by 42, 45, 48 and 52% respectively. The fungal antagonists reduce the grey mold disease severity between 5-42%. All treatments caused a decline in post harvest disease, as the most effective treatment of chemical control was benomyl with 68.33% and for the biological treatment this was T. koningii (T21) with 56%.

Biocontrol of Phytophthora cactorum the causal agent of root and crown rot on apple (Malus domestica) by formulated Pseudomonas fluorescens.

In this study 284 isolates were isolated of apple trees' rhizosphere from Iran and 128 isolates were obtained from the collection of Research Department of Biological Control of University of Tehran. Four strains (P60, P61, P96, and P97) of Pseudomonas fluorescens were selected for greenhouse trials. The results of greenhouse trials showed dipping the crown and root of apple seedlings (MM106) combined with soil drench was more effective than dipping the crown and root on reducing the disease. After 6 weeks, strain P60 in dipping method combined with soil drench with 70% control, exhibited greatest effect on reducing the crown and root rot and was more effective than the fungicide metalaxyl-mancozeb. After 12 weeks, strains P60 and P96 in dipping method combined with soil drench with 55.6% and 44.5% control respectively, exhibited greatest effects on reducing the diseases Study of media on growth rate populations of effective strains exhibited that the beet molasses yeast extract (1:1) had more effect than nutrient broth(NB) medium. The initial high populations of powder formulations of strains P60 and P96 decreased during the storage at 4 and 25 degrees C over a 150-day period. In addition, formulations of strains stored at 4 degrees C had longer shelf life than those stored at 25 degrees C. In glasshouse trials, after 6 weeks, formulation of strain P60 and unformulated P60, obtained from NB medium and formulated P60, obtained from molasses yeast extract medium, and metalaxyl-mancozeb had highest effect on reducing the disease on apple rootstocks. After 12 weeks, formulation of strain P60 and unformulated bacteria obtained from both media, and metalaxyl-mancozeb with 57.1% control showed greatest effect on reducing the disease.

Evaluation of production of some antifungal metabolites by fluorescent pseudomonads in vitro and their inhibitory effect on Penicillium digitatum in semi-commercial condition.

Epiphytic fluorescent Pseudomonads isolated from the surfaces of citrus leaves and fruits collected from the citrus orchards located in the north of Iran were screened for antagonistic activity against Penicillium digitatum caused agent of green mold of citrus fruit. 9 isolates revealed antagonistic effect in dual culture and then evaluated for production of antimicrobial metabolites such as production hydrogen cyanide on King's B medium containing glycine, production of siderophore on blue CAS-agar medium and production protease enzyme on Skim Milk Agar (SMA) medium. About 28% strains produced hydrogen cyanide, 83% strains produced siderophore and 70% of them produced protease. Antagonistic effects of the screened 9 isolates were studied by inoculating of the infected orange fruits with the bacterium at the concentration of 10(8) cells/ml under sterilized condition at 20 degrees C. The three final screened antagonist isolates were more studied in semi commercial trails in cold storage with dipping of the orange fruits in the bacterial suspension at the concentration of 10(8) cells/ml and then 5 weeks storage at 4 degrees C. The isolate P39 showed to be the most effective for controlling of this disease and decreased the damage by 77.28% respectively.

Effect of timing of application Pseudomonas fluorescens in suppression Sclerotinia sclerotiorum, the causal agent of white mold in canola.

Biological control of sclerotinia disease, as an important alternative to chemical control, has received considerable attention due to the lack of resistant varieties in most crop, and increasing concerns over fungicide resistance in population of Sclerotinia sclerotiorum, and fungicide residues in the environment. One biocontrol agent, Pseudomonas fluorescens PB-3, has been showed the antagonistic relationship between itself and S. sclerotiorum was investigated in this study. A petal infection technique was used to detect efficacy of timing of application strain PB-3 in the suppression of S. sclerotiorum on canola. Significant difference in disease severity (p<0.05) were found with respect to timing of ascospore applications in the control treatments (ascospores only). The superior competitive ability strain PB-3 was demonstrated by its complete suppression of disease severity when applied as a co-inoculation treatment or prior to ascospores inoculation. Analysis of effect of applying strain PB-3 after ascospores was indicated that treatment in which strain PB-3 was added to petals 48 or 24 h after ascospores, or when there were no bacteria present at all, had higher rates of disease progression. It would be appear that bacteria are able to significantly inhibit disease when applied before or even at the same time as ascospores. In a practical sense, this could mean that a field application of antagonist could be concurrent with infection by the pathogen.

Effect of carbon and nitrogen sources on growth and biological efficacy of Pseudomonas fluorescens and Bacillus subtilis against Rhizoctonia solani, the causal agent of bean damping-off.

One of the most important environmental factors that regulate the growth and antagonistic efficacy of biocontrol agents is the medium. The aim of this paper was to find the nitrogen and carbon sources that provide maximum biomass production of strains P-5 and P-6 (Pseudomonas fluorescens), B-3 and B-16 (Bacillus subtilis) and minimum cost of media, whilst maintaining biocontrol efficacy. All of the strains were grown in seven liquid media (pH=6.9) including: sucrose + yeast extract, molasses of sugar beet + yeast extract in 2:1 and 1:1 w/w ratios, molasses of sugar beet + urea, nutrient broth, molasses and malt extract, at an initial inoculation of 1 x 10(5) CFU ml(-1). Cells from over night cultures used to inoculate soil at 1 x 10(9) CFU cm(-3) soil. At the same time, fungal inoculum (infected millet seed with Rhizoctonia solani) was added to soil at the rate of 2 g kg(-1) soil. Results indicated that growth of P-6, B-3 and B-16 in molasses + yeast extract (1:1 w/w) medium was significantly higher than in the other media. Molasses + yeast extract (1:1 and 2:1 w/w) media supported rapid growth and high cell yields in P-5. In greenhouse condition, results indicated that the influence of the media on the biocontrol efficacy of P-5, P-6, B-3 and B-16 was the same and Pseudomonas fluorescens P-5 in molasses and malt extract media reduced the severity of disease up to 72.8 percent. On the other hand, there were observed significant differences on bean growth after one month in greenhouse. P-5 in molasses + yeast extract (1:1 w/w) medium had the most effects on bean growth promotion. In this study molasses media showed good yield efficacy in all of the strains. The high sucrose concentration in molasses justifies the high biomass in all of the strains. Also, the low cost of molasses allows its concentration to be increased in media. On the other hand, yeast extract was the best organic nitrogen source for antagonist bacteria but it is expensive for an industrial process. So it should be replaced by another industrial product instead of yeast extract, which confirm by an economic and technological study. The results obtained in this study could be used to provide a reliable basis to increase the population of biocontrol agents in fermentation process.

Biological control of Sclerotinia sclerotiorum (Lib.) de Bary, the causal agent of white mold, by Pseudomonas species on canola petals.

Sclerotinia sclerotiorum is an important pathogen on canola. Due to the public concern over pesticide use, alternative methods of disease control, such as biological control, should be considered. Several bacterial strains were isolated from canola and soja plants. Inhibition of S. sclerotiorum by bacterial strains in vitro was assayed on PDA medium in dual culture test. Eight Pseudomonas sp. strains (PB-3, PB-4, PB-5, PB-6, PB-7, PB-8, PB-10 and PB-11) caused inhibition zone against 5. sclerotiorum hyphal growth. The biocontrol potential of the bacteria was tested in a plant assay. Disease suppression was investigated using a petal inoculation technique. Canola petals were pretreated with bacteria, and then inoculated with 5. sclerotiorum ascospores 24 h later. Greenhouse experiment showed that application of Pseudomonas sp. strains (1 x 10(8) cfu ml(-1)) effectively suppressed S. sclerotiorum (1 x 10(5) ascospores ml(-1)) on petals and all of them achieved significant (P<0.01) disease suppression. Fourteen days after inoculation, strain PB-3 had 88/7% disease control and strain PB-4 had 69/9% disease control. Result from all studies indicates PB-3 to be effective biocontrol against S. sclerotiorum of canola. PB-3, PB-4, PB-7, PB-8, PB-10 and PB-11 were identified as Pseudomonas fluorescens biovar III. PB-5 and PB-6 was identified as Pseudomonas fluorescens biovar II. Strains PB-3, PB-4, PB-6, PB-10 and PB-11 produced protease and HCN. Strain PB-5 produce protease; no HCN.

Powder formulations of two strains of Bacillus subtilis for control of rape seed damping-off caused by Rhizoctonia solani.

Talc-based formulations of Bacillus subtilis strains B1 and B2 were tested as seed and soil treatments separately for their ability to control Rhizoctonia solani, the causal agent of rape seed damping-off, in greenhouse and field trials. In general, the formulated bacteria was more effective to suppress the disease than the suspension of bacterial cells in carboxymethylcellulose solution (1%, w/v), in both greenhouse and field trials. The formulations of strain B1 as soil treatment and strain B2 as seed treatment in greenhouse, and the formulations of strain B2 as seed and soil treatments in field trials had the greatest effect on reducing the rape seed damping-off (66.7%, 73.3%, 41.3%, and 42.4%, respectively). The formulations of strain B1 as soil treatment and strain B2 as seed treatment were the most effective treatments to increase the root dry weights in the infected soil in greenhouse. The formulation of strain B2 as soil treatment had the greatest effect on enhancement of the fresh weight of roots and stem fresh and dry weights. The formulations of strains B1 and B2 stored at 4 degrees C exhibited better shelf life and efficacy in vitro than their counterparts stored at 25 degrees C. Long-term stability of the formulation of strain B1 was found to be better.

Suppression of seed rot and preemergence of chickpea by seed treatments with fluorescent pseudomonads in Iran.

Species of Pythium isolated from rotted chickpea seeds and damped-off seedlings and chickpea soils at experimental field of Agriculture faculty of Tehran University in Karaj area that caused seed rot and preemergence damping-off of chickpea were Pyhium ultimum var. ultimum. One of the most important of soilborne fungal pathogens of the chickpea in Iran is seed rot and preemergence damping-off caused by Pythium ultimum Trow. Consequently, growers can expect as much as > 80% reduction in stand and yield if measures are not taken to control Pythium. Currently, most commercial seeds of chickpea are treated with pesticides. Fluorescent pseudomonads applied to seed are known to reduce soilborne diseases of chickpea caused by Pythium spp. In this study rotted chickpea seeds and diseased seedlings and soil samples were collected from experimental field in Karaj. Soils and roots used as sources of bacteria were collected from field. Fluorescent pseudomonads were isolated by plating samples on S1 and King's Medium B (KMB). Bacteria were preserved in 0.1 M MgSO4 for long-term storage; and NAG (containing 2% glucose) slants and plates at 4 degrees C short-term storage. Of 20 fluorescent pseudomonads isolated on S1 medium, 2 isolates selected for next tests. All strains significantly increased emergence as compared to the infested control in greenhouse trial; isolate Pf-4 consistently provided the best protection against Pythium. Seedling emergence from all bacteria seed treatments was statistically lower than the chemical treatments. All strains significantly increased fresh weight of chickpea as compared to the infested control in greenhouse trial. Seed treatment with metalaxyl were statistically better than captan in sterilized soil. In nonsterilized soil collected from the field artificially infested with P. ultimum, all strains significantly increased fresh weight of chickpea as compared to the infested control in greenhouse trial. Seedling emergence from seed treatment with isolate Pf-1 was the least effective treatment on fresh weight of chickpea in nonsterilized soil infested with P. ultimum. Seed treatment with captan was statistically better than metalaxyl in nonsterilized soil.

Biocontrol of Rhizoctonia solani, the causal agent of bean damping-off by fluorescent pseudomonads.

Rhizosphere bacteria belonging to the fluorescent pseudomonads are receiving increasing attention for the protection of plants against soil-borne fungal pathogens. Among these pathogens, Rhizoctonia solani, the causal agent of bean damping- off is very important in bean fields of Iran. In this study, the antagonistic activity of 46 isolates of fluorescent pseudomonads (isolated from different area of Iran) and Pseudomonas fluorescens strain CHA0 investigated against one isolate of R. solani. About 64% of isolates revealed antagonistic activity against R. solani. Production of antifungal metabolites such as HCN, siderophore and protease was evaluated. The results showed that 97.8%, 17% and 78% of isolates produced siderophore, HCN and protease respectively. There was no significant correlation between antagonistic activity and production of these metabolites. Isolates P-5, P-10 and P-32 with strain CHA0 were selected in order to investigate involvement of siderophore, volatile metabolites (HCN), and non-volatile metabolites in reducing mycelial growth of R. olani. Isolate P-5 showed much more inhibitory effect by production of volatile metabolites and siderophore. Non-volatile metabolites in isolates P-32 and P-5 completely inhibited mycelial growth of the fungus. After the primary labrotory tests, isolates P-14, P-35, P-30, P-5 and strain CHA0 were selected for in vivo experiments. These selected isolates with benomyl fungicide were used as seed coating and soil drenching in sterile soil under greenhouse condition. The result indicated that in seed treatment method, isolates P-30 by 66% had the most effect in disease reduction while in soil treatment method, strain CHAO by 60% had the most effect, such that this two isolates showed significant differences in comparison with plants inoculated with R. solani inoculums.

Contribution of phlA and some metabolites of fluorescent pseudomonads to antifungal activity.

Fluorescent Pseudomonas species are an important group of PGPR that suppress fungal root and seedling disease by production of antifungal metabolites such as 2,4-diacetylphloroglucinol (2,4-DAPG), pyoluteorin, pyrolinitrin, siderophores and HCN. The compound 2,4-DAPG is a major determinant in biocontrol of plant pathogens. A 7.2 kbp chromosomal DNA region, carrying DAPG biosynthetic genes (phlA, phlC, phlB, phlD, phIE and phlF). Detecting the ph1 genes make them an ideal marker gene for 2,4-DAPG-producing fluorescent pseudomonad's. In this study we detected ph1A gene (that convert MAPG to 2,4-DAPG) using PCR assay with primers phlA-1r and phlA- f that enabled amplification of phlA sequences from fluorescent pseudomonad's from ARDRA group 1 and 3. We could detect phlA gene in P. fluorescens strains CHAO, Pf-44, Pf-1, Pf-2, Pf-3, Pf-17, Pf-62 and Pf-64, native isolates of Iran. The efficacy of this method for rapid assay characterizing rhizosphere population of 2,4-DAPG producing bacteria from soil of different area of Iran is in progress. We used a collection of 48 fluorescent pseudomonas strains in vitro, with known biological control activity against some soil born phytopathogenic fungi such as, Macrophomina phaseoli, Rhizoctonia solani Vericillium dahlia, Phytophthora nicotiana, Pythium spp. and Fusarium spp. and the potential to produce known secondary metabolites such as protease. Strains Pf-1, Pf-2, Pf-3, Pf-17, Pf-33 and Pf-44 showed the best antifungal activity against all fungi used in this study. Thirty-eight of 48 strains produced protease. The ability to rapidly characterize populations of 2,4-DAPG producers will greatly enhance our understanding of their role in the suppression of root disease.

Biological control of Tiarosporella phaseolina the causal agent of charcoal rot of soybean.

Charcoal rot caused by Tiarosporella phaseolina (Tassi) Van der Aa is an important disease of soybean in Gorgan province of Iran. Experiments were carried out with 95 bactenal isolates that were collected from the rhizosphere of soybean plant. Among these bacteria only 50 isolates showed antagonistic effect on Tiarosporella phaseolina using dual culture test. Six highly effective bacteria were selected for subsequent studies. Based on biochemical physiological and morphological tests, isolates Pf-12 and Pf-63 were identified as Pseudomonas fluorescens, isolates B-13, B-42,B-126 and B-84 as Bacillus subtilis. The isolates of P. fluorescens produced antibiotics as well as volatile metabolites that inhibited mycelial growth of fungus. Bacillus subtilis isolates inhibited the fungal growth through volatile and non-volatile metabolites production. Only P. fluorescens isolates produced hydrogen cyanide. In greenhouse studies, the isolates B-13 and B-126 reduced 59% and 66% the intensity of charcoal rot of soybean respectively. The combinations of isolates B-13 and B-126 were also effective on reducing the intensity of disease.

Study on the overwintering of cleistothecia and conidia of Erysiphe betae causal agent of sugar beet powdery mildew in Iran.

Sugar beet leaves covered by sexual (cleistothecia) and asexual forms (mycelia and conidia) of Erysiphe betae were gathered at harvest time and maintained under natural outdoor conditions. In order to determine the function of cleistothecia and also conidia in the overwintering of E. betae some experiments were performed. The results showed that ascospores were unable to be released in petri dishes but their release under natural conditions occurred after 4 months. Under In vitro conditions ascospores did not germinate but on the leaves germination was rarely possible, however these ascospores were degraded after 7 days and didn't have pathogenicity. Conidia could induce pathogenicity after 3 but not 4 months. The period after inoculation until appearance of disease symptoms increased with aging of conidia. The results for conidial germination showed that fresh conidia had 80 percent germination on glass slides but it decreased sharply after 2 weeks and reached to 0 percent after 4 weeks. Although their germination on the leaves was not more than 46 percent of fresh conidia but they had good germination after 2 and 4 weeks. The results for the experiment to observe the first appearance of the disease in the field suggested that the first conidia were trapped by spore-trap in early June and the first symptoms appeared 20 days later. The conclusive results showed that ascospores had no function in the survival of the fungus and air-borne conidia are the main source of primary infections.

Occurrence and physico-serological properties of potato virus S (PVS) in Iran.

1818 collected samples of potato plant showing virus infection symptoms from 85 fields were tested for PVS infection using DAS-ELISA. Average of infection to this virus varied from 0 to 100%. Least infection was belonging to fields with new introduced varieties. On the other hand native and old introduced cultivars showed heavy infection. In field condition, PVS infected plants didn't show very obvious symptoms, so some infected plants may be missed in field sample collecting. The physical properties of 3 isolates, Avaj, Stanboly and Agria No 15 were determined. TIP 55-60 degrees C, DEP 10(-3) and Liv measured 3-4 days. Ouchterlony agar double diffusion test using SDS was useful for virus detection and precipitation lines didn't show any spur between isolates, although isolates differs slightly in symptomatology. SDS-Page and Western blotting methods used successfully for virus detection and determining and measuring viral protein components.

Biological control of chickpea Fusarium wilt by antagonistic bacteria under greenhouse condition.

In this survey, Fusarium oxysporum was isolated from roots infected plants and was shown to be pathogenic. Experiment were carried out with seven antagonistic bacteria. Based on biochemical, Physiological and morphological tests, isolates B-120, B-32, B-28 and B-22 were identified as Bacillus subtilis and isolates Pf-100, Pf-10 and CHAO as Pseudomonas fluorescens. In greenhouse studies, only isolate B-120 (Less than benomyl) reduced Fusarium wilt of chickpea in both seed and soil treatments. The application of antagonistic bacteria had no different effects on plant growth factors. Soil treatment of bacteria had a better effects on plant growth than that of bacterial seed treatment. The use of antagonists (B-120, B-28, B-120 and CHAO) in combination had no significant effect on plant growth factors and reduction wilt disease than that each isolate was applied individually.