New brevibacillus brevis strain, cultivation method and application of new brevibacillus brevis strain
A technology of Bacillus brevis and strains, applied in the field of microorganisms, can solve problems such as unsafe chemicals, and achieve the effects of good control effect, broad antibacterial spectrum and low cost
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Embodiment 1
[0038] Example 1: Isolation of Bacillus brevis strain GZDF3:
[0039] (1) Weigh 10g of Pinellia rhizosphere soil (5cm), pour it into a triangular flask filled with 90ml of sterile water, shake it on a shaker for 20min, then put it in a constant temperature water bath at 80°C and stir for 10min, and let it stand for 5min Draw 1mL for gradient dilution, the selected dilution is 10 -5 、10 -4 、10 -3 ;
[0040] (2) Take 100 μl of the soil dilution solution from step (1) and spread it on the NA medium plate, then place the plate upside down in an incubator at 28°C for 2-3 days, pick a single colony and inoculate it on a new medium for purification , repeated 3-4 times to obtain pure colonies;
[0041] (3) Antagonism test: adopt the agar punching method to evaluate the inhibitory effect of pinellia tuber rot: namely respectively apply the suspension of pathogenic bacteria and the suspension of pathogenic fungus spores evenly on the PDA plate, and then Each plate was punched with...
Embodiment 2
[0043] Embodiment 2: the preparation of biocontrol agent
[0044] (1) Strain activation: Take out the strain GZDF3 from the ultra-low temperature refrigerator at -80°C, streak it on the medium plate with an inoculation loop, and culture it at a constant temperature of 30°C for 28 hours, and grow a single colony;
[0045] (2) Preparation of seed solution: pick the above-mentioned single colony and inoculate it in 30 mL of NA liquid medium, shake it at 200 rpm at 30°C for 12 hours to obtain the seed solution;
[0046] (3) Preparation of liquid fermentation liquid: Inoculate the seed liquid in the fermentation medium at a ratio of 1:3.5 times of the inoculum and cultivate it. The initial pH is 7.5, the culture temperature is 32°C, and the shaking culture is 200 rpm for 36 hours, and then collected by centrifugation. Supernatant, stored as a biocontrol agent.
[0047] The strain medium is NA medium, and its components are: beef extract 5.0g / L, peptone 10.0g / L, sodium chloride 5.0...
Embodiment 3
[0050] Example 3 Control Effects on Pinellia tuber rot
[0051] Treatment method: There are 3 treatments in the test, treatment 1 is the sterile water control group, treatment 2 is Bacillus brevis GZDF3 bacterial agent, and treatment 3 is the commonly used fungicide 72% agricultural streptomycin sulfate (Chengdu Pratt & Whitney Bioengineering Co., Ltd. company) 500 times solution. Plant pinellia bulbs in the soil with pathogenic bacteria, among which treatment 2 soak the seeds with Bacillus brevis for 30 minutes before planting. When pouring the seedlings, use 1:50 times the bacterial agent to irrigate the roots once. In high temperature and high humidity weather, irrigate the roots once every 20 days. The treatment operations of the sterile water control group and the 72% agricultural streptomycin sulfate group were the same as above.
[0052] The results showed that the leaves of Pinellia ternata in the sterilized water control group turned yellow and withered, and the tub...
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