Bacillus amyloliquefaciens 2YN11 for disease prevention, growth promotion, quality improvement and adversity resistance of Lycopersicon esculentum, and application thereof
A technology of Bacillus amyloliquefaciens and tomato, applied to Bacillus amyloliquefaciens 2YN11 and its application fields, to achieve the effect of protecting the environment, promoting growth and promoting sustainable development
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Embodiment 1
[0027] Example 1 Screening method, morphological characteristics and physiological and biochemical characteristics of Bacillus amyloliquefaciens strain 2YN11 strain
[0028] Strain 2YN11 was obtained from soil screening from a farm in Jiangsu Province where tomato diseases were severe. The specific method is as follows. Using gradient dilution method, weigh 1g of soil from the diseased area and place it in a test tube containing 9mL of sterile water. Shake vigorously for 5 minutes to make a soil suspension. After 10-4 days of gradient dilution, spread on an LB plate and cultivate in the dark in a 28℃ incubator. After 48 hours, pick a single colony with different colony morphology, and store it in LB at 4℃ after being purified on the LB plate. The slope is ready for use.
[0029] The screened strains were subjected to plate antagonism experiments with several pathogens (bacterial wilt, bacterial spot, etc.). The specific method is as follows, taking bacterial wilt antagonism experi...
Embodiment 2
[0031] Example 2 Molecular identification of Bacillus amyloliquefaciens strain 2YN11
[0032] Bacterial Genome Kit (Cyberson) After extracting bacterial gene DNA, use gyrB universal primer gyrB1 (5'-GAAGTCATCATGACCGTTCTGCAYGCNGGNGGNAARTTYGA-3'); gyrB2R (5'-AGCAGGATACGGATGTGCGAGCCRTCNACRTCNGCRTCNGTCAT-3') to amplify the gyrB gene fragment from the genomic DNA . The specific amplification system and program (25ul) are as follows: primers 1, 2 (10μM) each 1μl; Mix: 12.5μl; Taq: 0.5μl. ddH 2 O: 10μl; template: 1μl. Pre-denaturation at 94°C for 5 minutes, denaturation at 94°C for 1 minute, annealing at 56°C for 2 minutes, extension at 72°C for 2 minutes, 35 cycles, extension at 72°C for 10 minutes, and storage at 4°C. The PCR amplified products were detected by 1% agarose gel electrophoresis, 90V, 30min; EB staining for 30min, the amplified products were detected. The amplified product was purified and ligated to the pMD19-T vector, transformed into Escherichia coli DH5α, extracted...
Embodiment 3
[0033] Example 3 Preparation of Bacillus amyloliquefaciens strain 2YN11
[0034] A kind of Bacillus amyloliquefaciens 2YN11 that can improve the biological control effect of various crop diseases, promote growth and resist stress, and contains microbial components: Bacillus amyloliquefaciens 2YN11 (preservation number 12606). It is isolated from the soil around the pear orchard in Zhejiang Province.
[0035] The preparation method of Bacillus amyloliquefaciens 2YN11 inoculum is as follows:
[0036] Bacillus amyloliquefaciens 2YN11 was cultured in LB broth at 28°C at 180rpm with shaking for 12-16h, and then centrifuged at 6000rpm for 10min to obtain the bacteria. The bacteria were diluted with sterilized water to make a microbe. The total concentration of viable bacteria in the product was 1×10 9 -1×10 10 CFU / mL.
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