Bacillus for controlling vegetable oomycetes disease and biological agent and application
A technology of biological preparations and bacillus, applied in the direction of biocides, chemicals for biological control, applications, etc., can solve the problems of increased residue, increased application amount, and decreased control effect, so as to promote proliferation and raw materials The effect of low cost and advanced production technology
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Embodiment 1
[0033] (1) Isolation of bacteria
[0034] Bacteria collected from the vegetable field soil in Wushan, Tianhe District, Guangzhou City, Guangdong Province were separated by plate dilution method, and the soil samples were diluted to 10 with sterile water. -6 、10 -7 、10 -8 、10 -9 、10 -10 Gradient concentration, NA solid medium (beef extract 3g, yeast extract 1g, peptone 5g, glucose 10g, agar 18g, water to 1000mL, pH 7.2-7.4) sterilized and poured into 9cm plates (about 15mL per plate ), use a pipette to pipette 0.1ml of each concentration into each dish, spread evenly with a coating rod, repeat 10 dishes for each concentration, cultivate at 30-32°C for 24-30h, and select each dish to grow 10-30 Pick a plate of 1 bacterial colony, aseptically suppress the slant of the NA test tube, incubate at 30-32°C for 24-30h, and store it for later use.
[0035] (2) Screening of bacteria for preventing and treating vegetable oomycosis
[0036] The bacteria isolated in step (1) are scree...
Embodiment 2
[0039] Taxonomic Identification of Bacteria PPRI 78
[0040] (1) Morphological and physiological and biochemical characteristics
[0041] Using NA medium plates, bacteria PPRI 78 was purified by dilution plate method and streaking method, and NA medium (beef extract 3g, yeast extract 1g, peptone 5g, glucose 10g, water to volume to 1000mL, pH 7.2-7.4) for 18h-24h, and Gram staining and observation of bacterial morphology and colony characteristics; refer to the method of Zhang Jizhong (1990) for aerobic test, carbon source utilization test, and indole test.
[0042] Single cell (0.6~0.8)×(2.2~3.0) μm, non-capsulated, perinatal flagella, Gram-positive bacteria, spore (0.7~0.9)×(1.0~1.4) μm, oval to columnar, located in the cell Central or slightly partial, the surface of the colony is rough and opaque, dirty white or yellowish, aerobic bacteria can use protein, xylose, fructose, sucrose, glucose and starch to decompose tryptophan to form indole.
[0043](2) 16S rDNA sequence a...
Embodiment 3
[0070] Example 3 Bacillus PPRI 78 Biological Preparation
[0071] (1) Activation of strains: After preparing NA medium (composition per liter: beef extract 3g, yeast extract 1g, peptone 5g, glucose 10g, water to 1000mL, pH 7.2-7.4), each specification Fill a 500mL Erlenmeyer flask with 100mL of NA medium, and sterilize at 121 for 15min. After cooling, insert Bacillus PPRI 78 (1 ring of cultured bacteria on the plate), put it into a shaker for fermentation and culture, the temperature is 28°C, the rotation speed is 200 rpm, and the culture time is 16 hours to obtain an activated bacterial liquid;
[0072] (2) Preparation of seed liquid: use a 100L fermenter to ferment, and the fermented liquid is LA medium (the composition of each liter is: cornstarch 3g, soybean protein powder 15g, corn steep liquor 25g, MnSO 4 0.0308mg, K 2 HPO 4 3g, KH 2 PO 4 1.5g, MgSO 4 0.5g, FeSO 4 0.1g, CaCO 3 1g, water is fixed to 1000mL, pH7.0), and liquid filling volume is 70L, and the b...
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