Biocontrol Strain bf6 for Controlling Tomato Yellow Leaf Curl Virus and Its Application
A technology of tomato yellowing curved leaves and bio-control strains, applied in the field of agricultural microorganisms, can solve the problems of high cost, limited resistant materials, and difficulty in large-scale promotion, and achieve the effect of saving costs and reducing residues
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Embodiment 1
[0022] Screening method: Pseudomonas BF6 was isolated from tomato rhizosphere soil in tomato greenhouses in Gaoyou Town, Gaoyou City, Yangzhou City. Gently brush the soil attached to the tomato root surface with a soft brush, weigh 3g and add it to 27ml of sterilized 0.85% NaCl, oscillate on a shaker at 180rmp for 0.5h, let it stand for 5min, to obtain a 10-1 dilution, and absorb the supernatant Solution 0.1mL was diluted sequentially, and 10 -4 、10 -5 、10 -6 Apply 0.1 mL each of the three gradient dilutions on the LB plate, incubate at 28°C for 48 hours, pick a single colony and inoculate it into fresh LB solid medium, and obtain purified strains through repeated transfer. The purified strains were stored in 40% glycerol in a -70°C ultra-low temperature freezer.
[0023] Identification method: The strain was identified through morphological characteristics, physiological and biochemical experiments and 16S rDNA sequence analysis.
[0024] Morphological characteristics: Gr...
Embodiment 2B
[0031] The preparation of embodiment 2BF6 biocontrol agent
[0032] After inoculating Pseudomonas BF6 (CGMCC NO.7238) into LB culture medium for 16 hours at 30°C with shaking at 200rpm, take samples in the ultra-clean workbench every 2 hours to measure its OD value at 600nm. When the OD value is between 0.5- At 0.8 o'clock, the cultivation was finished, and this bacterial solution was used as the seed bacterial solution. Inoculate the seed bacteria liquid into the LB fermentation culture medium at a volume ratio of 1:500 for fermentation and culture, and cultivate at 30°C and 200rpm for 24 hours. The pH value of the fermentation liquid is maintained at 7.0, the dissolved oxygen is 20%, and the ventilation rate is 5-7m 3 / hour, tank pressure 0.05-0.1KPa. Then centrifuge at 6000rpm for 10min, take the precipitate and dilute it with sterilized water to make bacterial preparation, the total concentration of viable bacteria in the finished bacterial preparation is 1×10 9 -1×10 ...
Embodiment 3
[0033] Example 3 Greenhouse Test Measures the Control Effect of BF6 on Tomato Yellow Leaf Curl Virus Disease
[0034] The tomato variety is "He He 903", which is transplanted at the 4-leaf stage and sprayed 5 days after transplanting. Get the bacterium preparation prepared in Example 2, dilute with water to the final concentration of bacterium liquid and be 5 * 10 7 CFU / mL, the spray standard is the distribution of mist droplets on each leaf, subject to not falling. The tomato yellow leaf curl virus was inoculated on the 7th day after the biocontrol agent treatment. The second same treatment was carried out 15 days after the first treatment. The treatment of inoculating virus alone without spraying biocontrol bacteria was used as the control treatment. There were 24 tomato plants in each treatment group, with three repetitions. The tomatoes were cultivated in a greenhouse with a temperature of 28°C and a light of 12h / 12h. When the disease was detected, the disease grade wa...
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