Burkholderia sp., microbial agent containing Burkholderia sp., bacterial fertilizer, preparation method and application
A technology of Burkholderia and Holderia, applied in the field of microbial preparations, can solve problems such as destruction, environmental pollution of ecosystems, and drug resistance of pathogenic bacteria
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[0030] The present invention also provides a preparation method for the bacterial agent described in the above scheme, comprising the following steps:
[0031] The Burkholderia bacteria are inoculated in LB liquid culture solution, and fermented to obtain bacterial agent.
[0032] In the present invention, the conditions of the fermentation culture preferably include: the OD value of the seed liquid of Burkholderia is 1.2, and the inoculum size of Burkholderia is the volume percentage content of the LB liquid culture solution 5% to 10%, the pH of LB liquid culture solution is 7.0, the culture temperature is 28 to 30°C, the shaking speed is 180 to 200 rpm, and the culture time is 60 to 80 hours. In the present invention, the culture time is preferably 72 hours.
[0033] The present invention also provides a bacterial fertilizer, which includes organic fertilizer and the bacterial agent described in the above scheme or the bacterial agent obtained by the preparation method.
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Embodiment 1
[0041] Isolation, screening and identification of embodiment 1 Burkholderia K2h bacterial strain
[0042] Isolation of strains
[0043] Test materials: Collected from the junction of soybean rhizomes planted in the suburbs of Longhai, Zhangzhou, Fujian, China.
[0044] Take the junction of edamame rhizomes, soak it in 75% alcohol for 30s on the ultra-clean workbench, rinse it with sterile water for 3 times, then soak it with 0.1% mercuric chloride for 1min, soak it with 75% alcohol for 30s, and rinse it with sterile water for 5 times. Weigh 1 g of surface-sterilized tissue block, soak it in 10 mL of phosphate buffer solution, grind it in a sterile mortar, and let it stand for 5 minutes. Take the supernatant and make a 10-fold gradient dilution with sterile water, and take 100 μL of the original solution and the diluted solution respectively. Spread the LB medium plate and incubate at 28°C for 2-3 days until a single colony grows. After the colony grows out, strains are purif...
Embodiment 2
[0055] The preparation of embodiment 2 biocontrol Burkholderia K2h microbial agent
[0056] The Burkholderia K2h stored at 4°C was activated on LB plate medium, and cultured at 28°C for 24 hours to obtain the activated strain. The activated strain K2h was picked with an inoculation loop and inoculated in 100 mL of LB culture solution, and cultured at 28° C. and 220 rpm for 24 hours to prepare a seed solution. Inoculate the Burkholderia K2h seed liquid (the OD value of the Burkholderia K2h seed liquid is 1.2) into the LB liquid culture liquid at an inoculum size of 5% by volume fraction, at 28° C., at 180 rpm After culturing for 72 hours, the fermentation broth of bio-control bacteria can be obtained, which is the bio-control bacterial agent. The total concentration of viable bacteria in the fermented bacterial agent is 8×10 8 ~5×10 10 CFU / ml.
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