Development of a selective medium for the determination of the spore concentrations of Botrytis cinerea in the air.
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Gray mold, caused by Botrytis cinerea, is an important disease that causes world-wide extensive damage to a wide range of economically important crops. When it is necessary to determine the spore concentration of Botrytis cinerea in a certain area, it is important to develop a method that can capture the spores of Botrytis cinerea and that can identify them. For the identification and enumeration of the plant pathogen Botrytis cinerea in the environment the easiest method available for the moment is the use of a selective medium. Several selective media for the isolation of Botrytis spp. have been developed by other research groups. All these media contain fungicides that are usually non-toxic towards Botrytis species and tannic acid, which is oxidized to produce a brown pigment that visualises the growth of Botrytis cinerea on the selective media. It seemed that different isolates of Botrytis cinerea that are found in nature have different sensitivities towards the different fungicide concentrations that are used in the selective media. Making the "optimal" selective media for Botrytis cinerea, we have to take in consideration that so many as possible Botrytis cinerea isolates must be able to germinate and grow on this selective medium and that the contamination of other micro-organisms on the selective medium must be minimized. Before the final composition of our selective medium for Botrytis cinerea, different combinations of fungicide concentrations were tried out of the following three fungicides: Rubigan, maneb and PCNB (pentachloronitrobenzene). All these selective media with different fungicides concentrations were tested out for spore germination and mycelium growth of Botrytis cinerea. Because it was obvious that the percentage Botrytis cinerea that germinated on PDA (Potato Dextrose Agar) was higher than on the selective medium a few experiments were executed in which the percentage of spore germination on PDA was compared with the percentage of spore germination on selective media. From the results of these experiments a correction factor was calculated that will be used when spore concentrations have to be determined for air detections that are carried out in glasshouses.Keywords:
Botrytis
Spore germination
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Endophytic actionmycetes No.37 was isolated from surface-sterilized tomato plant tissues.It had strong antifungal activity to Botrytis cinerea and other plant pathogens.The metabo-lic products of strain No.37 could strongly inhibit the mycelium growth and spore germination of Botrytis cinerea.The protective efficacy and therapeutic efficacy against Botrytis cinerea on pot tomato plants got to 89.7% and 80.3%,respectively.The effect of 10 times dilution against Botrytis cinerea was 84.1% in the field.
Botrytis
Spore germination
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Botrytis cinerea infects stems and leaves of greenhouse tomatoes and can cause serious economic losses. This study was conducted to develop environment-friendly control method against tomato gray mold. Antagonistic microorganisms (bacteria) were screened for control activity against Botrytis cinerea, both in vitro and in vivo, using stem sections. One hundred bacterial strains were isolated from the rhizospheric soil of various plants including tomato. These strains were screened for growth inhibition of Botrytis cinerea on agar plate by the dual culture and thirty strains showing strongly inhibitory effect against the pathogen were selected first. Among thirty strains, JB 5-12, JB 22-2, JB 22-3, U 4-8 and U46-6 reduced significantly disease incidence, when applied simultaneously with the pathogen. These results suggested that five antagonistic bacteria strains selected have the potential to control tomato gray mold in organic farming.
Botrytis
Plant disease
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To reduce the use of chemical agents, that are causing damage to the environment, in the fight against Botrytis cinerea, different BCA's were tested for their possibility to control Botrytis cinerea in a biological way. In order to investigate the effectiveness of the different micro organisms and Elot-Vis, experiments were carried out in climate chambers with 5 weeks old tomato plants. Leafs on the plant were inoculated with drops of a suspension that contained spores of Botrytis cinerea. The possible antagonists that were tested in these experiments were Trichoderma harzianum (Trichodex), T. asperellum (Biofungus), T. hamatum (T382), Bacillus subtilis (Serenade and Phytovit) and Pseudomonas aeruginosa (7NSK2 and KMPCH). For all these different micro organisms the direct and the indirect influence on Botrytis cinerea was investigated. In tests where the direct influence of the antagonists was examined, the spores of the moulds or the bacteria were suspended together with spores of Botrytis cinerea and subsequently drops of this suspension were pippeted on the leafs. After a while by ideal circumstances for Botrytis cinerea the infections on the inoculated leafs were counted. For the indirect influence of the antagonists, also leafs of 5 weeks old tomato plants were inoculated with a suspension of Botrytis cinerea spores. The roots of the tomato plants that were used for testing the indirect influence were treated during there growth with a suspension of the antagonist to see if induced systemic resistance pathway (ISR) was activated. For testing the effectiveness of Elot-Vis, tomato plants were sprayed a few times with a solution of this product during their growth. Results of the climate chamber test of the plants that were treated with Elot-Vis, showed a reduction of Botrytis cinerea infections on the inoculated leafs. Biological control agents seem to be not always very effective against Botrytis cinerea. The biological control agents that are containing micro organisms are also depending on the circumstances of the environment for an optimal development. These conditions are not always that optimal to compete with Botrytis cinerea or other micro organisms that are present on the plant.
Botrytis
Trichoderma harzianum
Trichoderma
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The present work aims to isolate actinomycete bacteria with antagonistic abilities towards Botrytis cinerea, the causal agent of gray mold, from a soil sample collected from the rhizosphere of a healthy tomato grove. In vitro confrontation led to the isolation of 104 actinomycete isolates; fifteen isolates have shown the most significant mortality rate of the mycelial growth of B. cinerea (>50%). Based on the results of this screening, representative strains were selected to verify their in vivo antagonistic activity on tomato fruits; the reduction of B. cinerea has a percentage ranging from 52.38% to 96.19%. Furthermore, the actinomycete isolates were evaluated for their plant growth-promoting (PGP) properties and their ability to produce biocontrol-related extracellular enzymes viz., amylase, protease, cellulase, chitinase, esterases, and lecithinase. Indeed, Ac70 showed high β-1,3-glucanase activity and siderophore production (17U/ml and 43% respectively), and the highest chitinase activity (39μmol/ml) was observed for Ac24. These results indicated that these actinomycetes might potentially control gray mold caused by B. cinerea on tomato fruits. Investigations on enhancing the efficacy and survival of the biocontrol agent in planta and finding out the best formulation are recommended for future research.
Chitinase
Lecithinase
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Isolate T95 of Trichoderma sp. Isolated from soil has shown the antagonistic action against the pathogen of Botrytis cinerea Pers. In solid culture medium. Antagonistic action showed parasitism and competition between isolate T95 and the above pathogen. Effective were 6×106/ml conidia ; The control experiment in the fields showed when effective inhibitory concentrations of isolate T95 were 5×106/ml conidia,it was the most effective in greenhouse experiments to against Botrytis cinerea Pers. and it′s control efficiency was more than 80.9%.
Trichoderma
Botrytis
Fungal pathogen
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The writer investigated and sampled grape grey mold in different regions of Xinjiang.According to its symptom,morphologic characters,cultural characteristics, host range,it was shown that the pathogen was Botrytis cinerea Pers.The temperature for this pathogen growth was from 5℃ to 30℃,with the optimum from 15℃ to 25℃;the pH values ranged from 2 to 9,with the optimum from 3 to 6;the different light had no significant effect on mycelial growth;this pathogen had wide range of hosts;Munage and Jufeng had high resistance to Botrytis cinerea;Thiophanate-methyl and Yijiale significantly inhibited the pathogen growth.
Fungal pathogen
Botrytis
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The strains antagonistic against Botrytis cinerea Pers.ex Fr.obtained from the sample of soil and diseased fruits of tomato in greenhouse have been screened in many ways.Results showed that there were only a few active strains among 165 isolated bacteria and 93 fungi strains,while about 20% strains have antagonism against the pathogen among 210 actinomycetes strains.There were 2 strains of bacteria(x24?x28),2 strains of fungi(z45,z120) and 3 strains of actinomycetes(f15,f111,f163) had good effects controlling the disease of Botrytis cinerea in vitro leaf screening test.
Botrytis
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A fungal peptide (from Saccharomyces cerevisiae -'brewers yeast') has been shown to be an inhibitor of fungal infection structures in the apple pathogens Venturia inaequalis (black spot or scab) and Glomerella cingulata (the bitter rot pathogen) in in vitro studies.Crude yeast extract also inhibits infection structure formation, although possibly by a different process.Both peptide and yeast extract were investigated for activity against V. inaequalis spores during infection of leaves.The effect of the peptide and crude yeast preparations on ascospore production was also investigated.Preliminary trials showed these fungal products had a major impact, strongly inhibiting sexual reproduction of V. inaequalis in vitro.The effect was most dramatic with the crude yeast extract treatments, with pseudothecium and ascospore production virtually eliminated.The peptide alone also had a major effect in reducing sexual spore production of the fungus.The effect of yeast extracts and pure peptides on the development of pseudothecia is under further investigation.
Botrytis
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