Bacillus amyloliquefaciens, microbial agent as well as preparation method and application of microbial agent
A technology of amylolytic spores and bacillus, applied in the field of microorganisms, can solve the problems of environmental pollution by chemical agents, unsatisfactory control effects, and drug resistance of pathogenic bacteria, and achieve good control effects, good stability, and high spore production.
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Embodiment 1
[0044] The isolation and screening of embodiment 1 bacterial strain F028
[0045] In the tomato planting field in Weifang City, Shandong Province, the surface soil was shoveled off, and about 500g of soil at a depth of 5-10cm was taken, put in a sterilized ziplock bag, and brought back to the laboratory. Strains were isolated by conventional dilution plate method, and antagonistic strains were screened with the pathogen of tobacco red spot disease as indicator bacteria. As a result, a strain with strong antagonistic effect was obtained, which was named F028.
Embodiment 2
[0046] Identification of embodiment 2 bacterial strain F028
[0047] 1. Morphological Identification
[0048] The strains were inoculated on NA plates, incubated at a constant temperature of 30°C for 12-48h, and the characteristics of the colonies were observed. The strains cultured for 12 hours were picked for Gram staining, and the strains cultured for 48 hours were picked for spore staining. The results showed that the colony was colorless and transparent at the initial stage, with a smooth, moist, sticky surface and irregular edges, and in the later stage, it was milky white with obvious bulges and wrinkles on the surface. Bacteria rod-shaped, spore-forming, Gram-positive. Such as figure 1 shown.
[0049] 2. Physiological and biochemical identification
[0050] Refer to "Bergey's Bacterial Identification Handbook" and "Common Bacterial System Identification Handbook" edited by Dong Xiuzhu and Cai Miaoying to conduct physiological and biochemical measurements on the sc...
Embodiment 3
[0062] The cultivation of embodiment 3 bacillus amyloliquefaciens F028
[0063] (1) Selection of medium
[0064] On the basis of 0.8% glucose, 1.5% corn flour, 1.5% soybean flour and 1000mL distilled water in the basic medium, the carbon source, nitrogen source and inorganic salt were optimized and screened by single factor test.
[0065] As carbon source, sucrose, maltose, lactose, soluble starch, mannitol, glycerin, citric acid and dextrin were used instead of glucose. The screening results showed that the strain cultured with mannitol as the carbon source had the highest biomass and the bacterial cell content was the highest. 1.1×10 9 CFU mL -1 , the spore content was 8.9×10 8 CFU mL -1 . Therefore, mannitol was finally determined to be the most suitable carbon source for F028 growth.
[0066] Nitrogen source, peptone, tryptone, urea, ammonium sulfate, ammonium nitrate, potassium nitrate, ammonium chloride and beef extract were added to the basal medium respectively. ...
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