A methylotrophic Bacillus and its application
A methylotrophic, bacillus technology, applied in applications, bacteria, fungicides, etc., can solve problems such as few disease-resistant varieties, poor chemical pesticide control effects, and environmental pollution
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Embodiment 1
[0027] Example 1 Isolation of methylotrophic bacillus TA-1
[0028] The maize rhizosphere soil collected from Tai’an City, Shandong Province in September 2014 was stored at 4 °C for isolation of antagonistic bacteria.
[0029] Weigh 10 g of sieved soil sample, put it into a 250 mL sterilized Erlenmeyer flask filled with 90 mL of sterile water and a small amount of glass beads, place the Erlenmeyer flask on a shaker at 37 °C and 200 r / min for 20 min, and make Suspend it sufficiently, then treat it in a water bath at 80°C for 10 min, and dilute the bacterial suspension to 10 with sterile water. -3 、10 -4 、10 -5 、10 -6 isogradient. Draw 100 µL of each gradient solution and evenly spread it on the LB medium plate, repeat each gradient 3 times, place in a constant temperature incubator at 30 °C for 1-2 days, pick different single colonies and purify by streaking on the plate. Store at 4°C until testing. Fusarium graminearum ( F. graminearum ), Rhizoctonia solani ( Rhizoct...
Embodiment 2
[0034] Example 2 Identification of Methylotrophic Bacillus TA-1
[0035] 2.1 Colony morphology and physiological and biochemical properties of strain TA-1
[0036] The activated strain TA-1 was streaked and inoculated on the NA plate, cultured at 37 °C for 24 h, and the growth, color and morphological characteristics of the colony were observed: the microscopic morphology of the strain TA-1 was rod-shaped, capable of producing spores, with Capsule. Normal growth on the NA plate, the single colony is irregularly round, with bulges on the edge, dry and wrinkled surface, opaque, and the colony is milky white ( figure 1 ); Bacteria formed when the LB liquid medium was static.
[0037] The indicators for the determination of physiological and biochemical properties mainly include Gram staining, VP (Voges-Proskauer) test, contact enzyme reaction, indole production reaction, salt tolerance, starch hydrolysis activity, casein hydrolysis test, citrate utilization test, methyl Red (...
Embodiment 3
[0051] Example 3 Pot control effect of methylotrophic bacillus TA-1 fermentation broth
[0052] Fusarium graminearum was inoculated into a 250 mL sterilized Erlenmeyer flask filled with 100 mL of CMC medium, and cultured on a shaker at 25 °C and 200 r / min for 3-5 days to obtain macroconidia. Remove the mycelium by filtering with sterile filter paper, collect the conidia suspension, and adjust its concentration to 10 with a hemocytometer. 5 Spores / mL. The acquisition of TA-1 fermented liquid is as embodiment 2, and gradient dilution becomes each concentration (10 8 -10 5 CFU / mL).
[0053] Firstly, corn seeds with full grains and the same size are selected, and sterilized with 1% NaClO. Then the corn seeds were soaked in different concentrations of TA-1 fermentation broth (10 4 -10 8 CFU / mL) for 2 h, air-dried overnight and sowed in nutrient pots (diameter 15×height 12 cm) filled with sterilized natural soil and seedling substrate, sowed 8 corn seeds in each pot, and soa...
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