Stable Burkholderia, bacterial agent and preparation method and application thereof
A technology of Burkholderia and Holderia, which is applied in the field of microorganisms, can solve the problems of less effective and poor microbial agents, increase crop yield, reduce ethylene production, and enhance stress resistance Effect
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Embodiment 1
[0031] Embodiment 1 Isolation and screening of stable Burkholderia FX-1 bacterial strain
[0032] (1) Weigh 1 g of the agricultural saline-alkali land soil (soil salinity is 4%) collected in Fengxian District, Shanghai (30°52′N, 121°34′E) and suspend it in 50mL PAF culture solution, at 30°C, 200r After 24 hours of culture on a shaker at 1 / min, the suspension was transferred to DF and ADF liquid medium and continued to culture for 24 hours. After the culture was completed, the bacterial solution was diluted by 10 4 、10 5 and 10 6 Then spread the diluted bacterial solution on the plate of LB solid medium with a coating rod. After a single colony grows on the plate, pick a single colony to separate and purify.
[0033] (2) Inoculate the strains screened in step (1) in TSB liquid medium respectively, culture them on a shaker at 30°C and 200r / min for 24 hours, then centrifuge at 8000rpm to get the bacteria, rinse with DF culture medium and resuspend in ADF in liquid medium. Th...
Embodiment 2
[0044] The preparation method of embodiment 2 stable Burkholderia bacteria agent
[0045] (1) In the ultra-clean workbench, use a pipette gun to draw 50-100 μL of FX-1 bacterial liquid into the sterilized plate containing TSB solid medium, and place it in a constant temperature incubator at 25-30°C for 18-24 hours ;
[0046] (2) In the ultra-clean workbench, use an inoculation loop to pick a single colony from the plate containing the TSB solid medium into 50 mL LB liquid medium for liquid fermentation to obtain a liquid fermentation broth. The fermentation conditions are: pH value 7.2-7.5, 30°C culture condition, 200rpm shaking culture for 16-19h.
[0047] (3) Measure the absorbance (λ=600nm) of the liquid bacterial solution. When the light absorption value of the bacterial solution is 1, dilute it 10 times to obtain 500mL bacterial agent, and the number of FX-1 active bacteria is about 10 8 individual / mL.
[0048] (4) Store the prepared bacterial agent in a refrigerator ...
Embodiment 3
[0052] Embodiment 3 stable Burkholderia bacterial agent (hereinafter referred to as FX-1 bacterial agent) indoor inoculation test
[0053] The model organism Arabidopsis thaliana was selected as the research object, and pot inoculation experiments were carried out in a constant temperature incubator. Arabidopsis seeds were sown in flower pots with a diameter of 5 cm and a height of 6 cm, 5 seeds were sown in each pot, and the sowing depth was about 1 cm. After that, sterile water was sprayed and placed in a constant temperature incubator with the temperature set at 25°C. The light period is 14h, and the dark period is 10h. After the emergence of Arabidopsis thaliana, keep one Arabidopsis with the best growth in each pot.
[0054] Two treatments were set up with FX-1 bacterial agent and no FX-1 bacterial agent applied, and the control group without the bacterial agent was applied, and each treatment had 3 replicates. Use a syringe to draw 1mL FX-1 bacterial solution, slowly i...
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