Disease-preventing, growth-promoting and stress-resisting bacillus amyloliquefaciens Y15 for all plants and application of bacillus amyloliquefaciens Y15
A technology for dissolving starch spores and bacilli, applied in plant growth regulators, plant growth regulators, applications, etc., to achieve the effects of improving stress resistance, improving quality, and promoting sustainable agricultural development
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Embodiment 1
[0026] Example 1 Molecular identification of Bacillus amyloliquefaciens strain Y15
[0027] Y15 was isolated from the rhizosphere soil of zhejiang pear orchard.
[0028] After extracting the bacterial gene DNA with the Bacterial Genome Kit (Saibaisheng Company), the gyrB gene fragment was amplified from the genomic DNA with gyrB universal primers gyrB1 (5'-GAAGTCATCATGACCGTTCTGCAYGCNGGNGGNAARTTYGA-3'); gyrB2R (5'-AGCAGGATACGGATGTGCGAGCCRTCNACRTCNGCRTCNGTCAT-3') . The specific amplification system and procedure (25ul) are as follows: 1 μl each of primers 1 and 2 (10 μM); Mix: 12.5 μl; Taq: 0.5 μl. wxya 2 O: 10 μl; template: 1 μl. 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. PCR amplified products were detected by 1% agarose gel electrophoresis, 90V, 30min; EB stained for 30min, and amplified products were detected. ...
Embodiment 2
[0030] The preparation method of Bacillus amyloliquefaciens Y15 bacterial agent is as follows:
[0031] Bacillus amyloliquefaciens Y15 was shaken and cultured in LB culture medium at 28°C and 180rpm for 12-16h, then centrifuged at 6000rpm for 10min to obtain the bacterial cells, and the bacterial cells were diluted with sterilized water to make bacterial agents. The total concentration of viable bacteria in the finished bacterial agent was 1×10 9 -1×10 10 CFU / mL.
Embodiment 3
[0032] Embodiment 3 Bacillus amyloliquefaciens Y15 greenhouse growth-promoting test
[0033] Method: use 10% H to test watermelon, tomato and cucumber seeds 2 o 2The surface was disinfected for 10 minutes, washed with sterile water and dried for later use. Sow 10 seeds per nutrient bowl. After 4 weeks of normal watering management, the seedlings with the same growth were transplanted into 330ml plastic cups. 7 The concentration of CFU / mL is applied to the roots of crops by irrigation, 30mL per plant. Then placed in a greenhouse for cultivation, 28°C, 70% relative humidity. The water treatment was used as the control, and 48 seedlings were treated in each group, placed randomly, and repeated three times. After 30 days, the plant height, stem diameter, fresh weight and dry weight of watermelon, tomato and cucumber were measured.
[0034] Use 10% H to test the seeds of rice and water spinach 2 o 2 The surface was disinfected for 10 minutes, washed with sterile water and d...
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