Enterobacter 3bh19 and its application to improve the microecology of cucumber leaf circle to control downy mildew
A technology of Enterobacteriaceae and Enterobacter strains, applied to Enterobacter 3bh19 and its application fields, can solve the problems of farmers' economic loss, reduced yield, and delayed fruit development, and achieve the effect of protecting the environment and promoting sustainable development.
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Embodiment 1
[0024] Example 1 Molecular Identification of Enterobacter 3bh19
[0025] Bacterial Genome Kit (Saibaisheng Company) After extracting bacterial gene DNA, use gyrB universal primers gyrB1 (5'-GAAGTCATCATGACCGTTCTGCAYGCNGGNGGNAARTTYGA-3'); gyrB2R (5'-AGCAGGATACGGATGTGCGAGC CRTCNACRTCNGCRTCNGTCAT-3') to amplify gyrB gene from genomic DNA fragment. 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 5min, denaturation at 94°C for 1min, annealing at 56°C for 2min, extension at 72°C for 2min, 35 cycles, extension at 72°C for 10min, 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. After purification, the amplified product was connected to the pMD19-T vector, transformed into Escherichia coli DH5α, and the positive clone plasmid ...
Embodiment 2
[0027] An Enterobacter 3bh19 that improves the biological control effect of cucumber downy mildew, promotes growth, resists stress, and improves microecology, and contains microbial components: Enterobacter 3bh19 (preservation number is CGMCC NO.12608). It was isolated from the rhizosphere soil of Huai'an cucumber.
[0028] The preparation method of Enterobacteriaceae 3bh19 bacterial agent is as follows:
[0029] Enterobacter 3bh19 was 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 then diluted the bacterial cells with sterilized water to make bacterial preparations. The total concentration of viable bacteria in the finished bacterial preparations was 1× 10 9 -1×10 10 CFU / mL.
Embodiment 3
[0030] Example 3 Greenhouse Growth-promoting Test of Enterobacter 3bh19 on Cucumber
[0031] Method: soak the cucumber seeds with 10% H 2 o 2 The 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 consistent growth were transplanted into 330ml plastic cups, and the suspended Enterobacter 3bh19 bacterial agent was mixed with 1.0×10 7The 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, dry weight and other biomass of cucumber plants were measured.
[0032] Table 1 Effect of Enterobacter 3bh19 on cucumber biomass
[0033]
[0034] Not...
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