Biocontrol bacillus subtilis WJ-1 for rice sheath blight, microbial inoculum and application thereof
A technology of Bacillus subtilis and rice sheath blight, applied in the direction of application, biocide, fungicide, etc., to achieve obvious control effect, simple production and use
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Embodiment 1
[0034] Example 1: Isolation and identification of Bacillus subtilis WJ-1 and research on its bacteriological characteristics
[0035] 1. Isolation and identification of Bacillus subtilis WJ-1
[0036]A biocontrol bacterium Bacillus subtilis (Bacillus subtilis) WJ-1 suitable for the prevention and treatment of rice sheath blight of the present invention was isolated and screened on the vegetable garden soil of Huazhong Agricultural University campus in Shizishan District, Wuchang, Wuhan City, Hubei Province. Its separation and identification methods are carried out according to the method reported (Edited by Zhou Deqing, "Microbiology Experimental Technology Manual", Shanghai Science and Technology Press, 1986 edition), which is a routine microbial isolation and identification method. The bacterial strain has been submitted to the Chinese Type Culture Collection Center (CCTCC) located in Wuhan University, Wuhan City, Hubei Province for patent procedure preservation on March 30,...
Embodiment 2
[0049] Embodiment 2: the preparation of bacillus subtilis WJ-1 bacterial agent
[0050] (1) Activation culture of the WJ-1 strain: First, the WJ-1 strain with the deposit number CCTCC NO: M2010069 was activated on a PDA medium (formulation as described in Example 1) plate at 33° C. for 1 day.
[0051] (2) Liquid culture of the WJ-1 strain: Inoculate the single colony prepared in step 1 into a 500ml Erlenmeyer flask containing 200ml of PDB (the formula is the same as PDA, except that no agar is added), 180r / min, 33°C shaking culture for 48h, that is It is fermentation broth A, wherein the concentration of Bacillus subtilis WJ-1 is 1.5×10 10 cfu / ml.
[0052] (3) Preparation of WJ-1 bacterial agent: Centrifuge the fermentation broth prepared in step 1 at 6,000r / min for 20min at 4°C, take the centrifuged supernatant, and filter it with a common bacterial filter (pore size 0.22μm) supernatant to obtain a sterile fermentation filtrate (abbreviated as fermentation broth B). Wash t...
Embodiment 3
[0053] Embodiment 3: In vitro and indoor living test of bacillus subtilis WJ-1 bacterial agent for controlling rice sheath blight
[0054] (1) Indoor in vitro inoculation control test:
[0055] The rice sheath blight strain WH-1 of the present invention was activated on a PDA medium plate at 28° C. (for the method, refer to the method reported by Xie et al., 2008). Cut 7.5 cm leaves of rice seedling stage (5-leaf stage), place them in a sterile glass petri dish (diameter 9 cm), put absorbent paper in the petri dish to keep moist, and each dish has 3 leaves. Setup processing includes:
[0056] ① Spray clean water, fermentation broth A, fermentation broth B and bacteria fluid C until there is a layer of liquid film on the surface of the rice leaves, and after 12 hours, connect the activated 5mm rice rhizoctonia (WH-1) bacteria block to the rice leaves , a distance of about 2cm;
[0057] ②Spray clear water, fermentation broth A, fermentation broth B, and bacteria fluid C until...
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