Biocontrol of Pseudomonas and its application
A technology of Pseudomonas and biocontrol strains, applied in the field of biochemistry, can solve the problem of single antibacterial action mechanism, achieve good control effect, convenient application method, and simple preparation process
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Embodiment 1
[0028] The isolation and screening of biocontrol strain GZ9, the specific steps include:
[0029] Step 1) Isolation and Preservation of Soil Pseudomonas
[0030] The biocontrol strain GZ9 was isolated from the corn rhizosphere soil in Bijie City, Guizhou Province. The isolation process was as follows: Weigh 10 g of the soil sample into a sterile Erlenmeyer flask, add 90 mL of sterile water, and sterilize 5 g of small steel balls at 28 ° C. , 180rpm shaker for 20min, and then stand at room temperature for 5min to obtain 10 -1 Diluent, absorb 100μL and add to 900μL sterile water, mix well to get 10 -2 Diluent, adopt the same method to carry out gradient dilution, obtain 10 -3 and 10 -4 Diluent. 100 μL of each gradient was evenly spread on the Pseudomonas selective medium, and 3 plates were plated for each gradient, placed in a 28°C incubator, and cultured upside down.
[0031] After 1 to 3 days, pick a single colony and grow on LB solid medium. After 1 day, select the strai...
Embodiment 2
[0045] The identification of biocontrol strain GZ9, the specific steps include:
[0046] Step 1) Physiological and biochemical characteristics
[0047] Table 2 The main physiological and biochemical characteristics of strain GZ9
[0048]
[0049]
[0050] Step 2) 16S rRNA gene identification
[0051] Use bacterial 16S rRNA gene universal primer 27 as shown in Seq ID No.4
[0052] F(5'-AGAGTTTGATCCTGGCTCAG-3') and 1492 as shown in Seq ID No.5
[0053] R(5'-TACGGCTACCTTGTTACGACTT-3'), using the genomic DNA of strain GZ9 as a template; carry out PCR reaction according to the following reaction system (30 μL): 2×Taq PCR StarMix with Loading Dye 15 μL, template DNA 1 μL, upstream primer 27F 1.4 μL , downstream primer 1492R 1.4 μL, ddH 2 O 11.2 μL. PCR reaction program: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 90 s, a total of 32 cycles; final extension at 72°C for 10 min. The PCR product is abou...
Embodiment 3
[0056] The preparation of the GZ9 biocontrol agent, the specific steps include: insert the biocontrol strain GZ9 into the LB liquid medium, cultivate it at 28°C and 180rpm for 12h, and make the seed liquid; inoculate the seed liquid with 1% inoculum in the fermentation culture medium, 28°C, 180rpm shaking culture for 48h, to obtain a fermentation broth, the total concentration of viable bacteria was 1×10 9 CFU / mL or more is the prepared biocontrol agent. The fermentation medium still uses LB liquid medium.
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