Anti-disease, growth promotion and drought-resistant functional plant endogenous bacillus velezensis and application thereof
A technology of bacillus and microbial strains, applied in forestry and horticultural crop protection, and agricultural fields, can solve problems such as differences, achieve rapid growth, short fermentation cycle, easy large-scale production and commercial application
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Embodiment 1
[0054] Example 1, the isolation and identification of Bacillus velezensis E6
[0055] (1) Isolation of Bacillus E6 strain
[0056] Bacillus velezensis (Bacillus velezensis) E6 of the present invention is obtained from the marigold root, after diluting and coating the tissue grinding liquid, separating and obtaining, and the separating method is:
[0057] (1) Isolation of Bacillus Velez: The roots of plants such as marigolds were collected at the experimental base of the Academy of Agricultural Sciences in Langfang City, Hebei Province. Weigh 1g root, wash it with 1% NaClO sterilized water, put it into a sterile mortar and grind it, use a pipette gun to draw 1mL grinding solution from the mortar, add it to another test tube filled with 9mL sterile water, and mix well , and so on to make 10 -3 、10 -4 、10 -5 Solutions of different dilutions. Take 100 μL of the diluted solution and spread it on a TSA medium plate, spread it on three culture dishes for each gradient, and incub...
Embodiment 2
[0074] Preparation of Bacillus velezensis E6 bacterial agent
[0075] 1. The bacterial lawn of Bacillus velezensis E6CGMCC No.9665 obtained in Example 1 was transplanted into nutrient broth (NB), and shaken at 28-30° C. for 20 hours to obtain seed liquid.
[0076] Among them, the formula of nutrient broth (NB) is: glucose 10g, peptone 10g, beef extract 3g, yeast extract 1g, distilled water 1000mL, pH adjusted to 7.0-7.2, autoclaved at 121°C for 30 minutes.
[0077] 2. Inoculate the seed liquid obtained in step 1 into a large amount of culture and fermentation culture liquid according to the ratio of 1% (volume ratio), the culture temperature is 30-32 ° C, the rotation speed is 150-180 rpm, the initial pH is 7.0-7.2, and the culture time is 28 hours . Fermentation liquid is obtained after the fermentation is completed, and the number of live bacteria of Bacillus velezensis E6CGMCC No.9665 in the fermentation liquid can reach 12 billion / mL.
[0078] 3, the bacterium slime obta...
Embodiment 3
[0079] Embodiment 3: the disease prevention effect of bacillus E6 bacterial agent
[0080] Control Effects on Cucumber Fusarium Wilt : Fusarium was cultured with Armstrong culture medium first, counted under the microscope after 3 to 4 days, and the bacterial solution was diluted to 5×10 4 cfu / mL for use; dilute the E6 bacterial agent to 1×10 7cfu / mL. Mix the Fusarium spore suspension and the E6 bacterial agent in equal volumes, then add the sterilized seeds into the mixture and soak for 30 minutes. There were 10 repetitions per treatment, and 3 cucumber seeds were sown in each repetition, and the treatment without fungal agent was used as the control. 3 weeks after sowing, the growth and disease of cucumbers were investigated.
[0081] The components and proportioning of the Armstrong culture solution are: glucose 20.0g, KCl 1.6g, MgSO 4 .7H 2 O0.4g, Ca(NO 3 ) 2 5.9g, KH 2 PO 4 1.1g, FeSO 4 0.2μg / mL, ZnSO 4 0.2μg / mL, MnSO 4 0.2μg / mL, distilled water 1000mL, aut...
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