Wheat scab liquid biocontrol agent and preparation method thereof
A technology for wheat scab and biocontrol fungicides, which is applied in biochemical equipment and methods, botanical equipment and methods, and microorganism-based methods, etc., can solve problems such as loss and production failure, and achieve good control effects.
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Embodiment 1
[0015] Example 1: Isolation and screening of bacterial strain LM3403
[0016] Collect wild yak dung that grows in the alpine and pollution-free grassland above 3,000 meters above sea level. The method of gradient dilution and coating separation is used to separate the bacterial strains in the sample by coating. Colonies with obvious morphological differences were picked for streak purification, numbering, and preservation for future use. The antibacterial ability of the tested strains was investigated by filter paper method and plate confrontation method, and the strain with the strongest broad-spectrum antibacterial ability was named LM3403. The width of the inhibition zone of strain LM3403 against Fusarium graminearum was (6.7±1.1) mm.
Embodiment 2
[0017] Embodiment 2: identification of bacterial strain LM3403
[0018] The colony of strain LM3403 on the NA plate is round, opaque and milky white, sticky, and the edges are irregular; the cells are rod-shaped, Gram-positive, spore-shaped, and oval. Aerobic growth: positive, motility: positive, V-P test: positive, catalase test: positive, indole test: positive, starch hydrolysis: positive, casein hydrolysis: positive, gelatin liquefaction: positive, glucose fermentation test: Positive, nitrate reduction: positive, urea utilization: positive, the morphological characteristics, physiological and biochemical characteristics of the strain LM3403 are consistent with the standard strain of Bacillus subtilis. The 16S rDNA sequence of strain LM3403 and Bacillus subtilis strain SPA N1(JQ308575), Bacillus subtilis strain ME2(JQ900623) and Bacillus subtilis The homology of strain BDT3(HQ711982) reached 100%. Comprehensive analysis determined that the strain LM3403 was Bacillu...
Embodiment 3
[0019] Example 3: Extraction and analysis of bacterial strain LM3403 antibacterial lipopeptide
[0020] Centrifuge the fermentation broth of the strain at 4°C and 12000rpm for 15min, collect the sterile supernatant; use 6mol / L HCl to adjust the sterile supernatant to pH2.0, let stand overnight at 4°C, and then centrifuge at 12000r / min at 4°C After 15 minutes, collect the precipitate; add an appropriate amount of methanol to the precipitate, stir on a magnetic stirrer for 30 minutes, put it in a refrigerator at 4°C for extraction for 8 hours, and centrifuge at 12,000 rpm for 15 minutes at 4°C to obtain methanol extract. After concentration under reduced pressure and vacuum freeze-drying, the antibacterial substance was obtained and stored at -20°C. The antibacterial substance after vacuum freeze-drying was taken, dissolved in phosphate buffer solution with pH 7.0, prepared into a certain concentration antibacterial substance solution, and stored at 4°C for later use.
[0021] ...
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