Bacillus velenzensis and application thereof to biological control
A technology for separating Bacillus leissi and Bacillus, applied to Bacillus veleis and its application in biocontrol, can solve problems such as yield loss of rapeseed black shank
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Embodiment 1
[0037] Screening and Identification of Bacillus Velez Bv-6:
[0038]The bacterial strain involved in the present invention is isolated from a rapeseed endophytic bacteria sample collected in Wuhan City, Hubei Province, and the bacterial strain Bv-6 has an obvious effect after antagonism testing and plant growth promoting testing. It was identified as Bacillus velezensis by morphology and molecular biology. The strain was sent to the China Center for Type Culture Collection on December 25, 2019. It was named after classification: Bacillus velezensis Bv-6 , the deposit number is: CCTCC NO: M 20191106 Address: Wuhan University, Wuhan, China.
[0039] Bacillus Velez belongs to Gram-positive bacteria. Its body is rod-shaped and its size is 0.5μm×(1.5~3.5)μm. Stress-resistant spores can be produced in the middle or at the tail, and the spores are oval. In three-section line culture of beef extract peptone (NA) medium, the single colony was milky white, with rough surface and irreg...
Embodiment 2
[0041] The preparation of the bacterial agent of Bacillus Velez Bv-6:
[0042] Fermentation broth preparation:
[0043] Streak inoculate the Bacillus velei Bv-6 preserved in glycerol at -80°C on the solid NA medium, culture it in a 30°C incubator for 24 hours, and then pick a single colony and inoculate it into 100mL of liquid NB culture In the medium, shake culture under the condition of 30 ℃ and 180rpm for 24h, as the 10L fermenter seed liquid of Bacillus veleisi Bv-6.
[0044] The formula of the fermentation medium is beef extract 3g / L, soybean peptone 5g / L, glucose 20g / L, pH 7.
[0045] Sterilize 6L of the above-mentioned fermentation medium in a 10L fermenter by high-pressure steam at 121°C for 0.5h, cool down to 40°C after sterilization, and immediately inoculate 1×10 7 300mL (5% inoculum size) of Bacillus velei seed solution at cfu / mL concentration, cultivated for 72 hours at a culture temperature of 30°C, a rotating speed of 300r / min, an air pressure of 0.05MPa, and ...
Embodiment 3
[0052] Determination of the Effect of Bacillus Velez Bv-6 on Leptosphaeria biglobosa (Lb) Infection on Rapeseed Leaves
[0053] Wash the conidia of the Lb strain with sterile water, put them in a 50mL centrifuge tube, filter through four layers of lens-cleaning paper to obtain a conidia suspension, and adjust its concentration to 1×10 8 spores / mL.
[0054] Preparation of powder suspension: get 3 g of the powder prepared in Example 2 and dilute it in 30 mL sterile water, and dilute to a concentration of bacteria equal to 1×10 7 cfu / mL to obtain a powder suspension.
[0055] On each true leaf of rapeseed (Zhongshuang No. 9), the Bacillus Veles Bv-6 fermentation liquid and powder suspension prepared in Example 2 are used to evenly smear on the surface (about 10 μ L of Bv-6 is added dropwise on each true leaf). fermentation broth, powder suspension). The leaves treated with NB solution were used as the control, and each treatment was repeated 4 times (the 4 repetitions here ref...
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