Bacillus safensis having growth promoting function, bactericide and application thereof
A technology of bacillus saphobia and microbial inoculants, applied in the field of microorganisms, can solve the problems that research results remain at the experimental level, the number of pro-growth inoculants are not large, and the research and development of high-efficiency fermentation processes and formulations are lagging behind.
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Embodiment 1
[0036] Isolation, identification and preservation of the bacterial strain of embodiment 1
[0037] 1. Isolation of AMCC101030 strain
[0038] A growth-promoting strain was screened out from plant rhizosphere soil and named AMCC101030 strain.
[0039] For the colony morphology and microscopic morphology of AMCC101030 strain, see figure 1 . in figure 1 The left image in the figure is a micrograph, figure 1 The right figure in the figure is a photo of the colony morphology;
[0040] Depend on figure 1 As can be seen from the right picture in the figure, the shape of the colony is irregular and circular, the edges are not neat, the surface is rough with many folds, opaque, the color of the colony is milky white, and the color of the culture medium has no obvious change. Microscopic observation of Gram staining was positive, the cells were rod-shaped, and the spores were oval. Its catalase, casein hydrolysis, gelatin hydrolysis, citrate utilization, D-glucose acid produc...
Embodiment 2A
[0044] Embodiment 2AMCC101030 bacterial strain produces IAA ability determination
[0045] Use an inoculation loop to pick the AMCC101030 strain preserved on the slant and streak it on LB (0.5% yeast extract, 1% peptone, 1% sodium chloride, 1.6% agar, pH7.0, 121 ° C, 20min high-pressure sterilization, all are mass percentages ) plate activation; the activated bacteria were inoculated in King culture medium (peptone 20g, glycerol 15.0g, K 2 HPO 4 1.15g, MgSO 4 ·7H 2O 1.5g, distilled water 1 000mL, pH 7.2, add 0.05% L-type tryptophan, 121°C, 20min autoclave), 37°C, 200rpm shaker culture for 6 days, 9000 rpm for 10min, collect the supernatant , add 2ml colorimetric solution (Fe-HClO 4 Composition: 1ml 0.5M FeCl 3 solution plus 50ml of 35% HClO 4 solution, mixed), reacted in the dark for 30min, and measured OD 530 value.
[0046] Preparation of the standard curve: Weigh the standard substance of indole-3-acetic acid (IAA), and prepare standard solutions with concentration...
Embodiment 3
[0050] The preparation of embodiment 3AMCC101030 bacterial agent
[0051] The cultivation method of Shafu bacillus (Bacillus safensis) AMCC101030, the steps are as follows:
[0052] (1) Streak inoculation of Bacillus suffolus AMCC101030 cryopreserved on an LB solid medium plate, and culture at 37°C for 24 hours to obtain an activated strain;
[0053] (2) Inoculate the activated bacterial strain prepared in step (1) in a conical flask equipped with LB liquid medium, and culture it in a constant temperature oscillator at 37° C. and 180 rpm for 18 hours to obtain a seed solution;
[0054] (3) Inoculate the seed solution prepared in step (2) into the liquid medium with an inoculum size of 8%, and cultivate it at 37° C. for 8 hours to obtain a primary fermentation solution;
[0055] (4) Inoculate the primary fermentation broth prepared in step (3) into the fermentation medium with an inoculum size of 5%, and culture at 37° C. for 24 hours to obtain a fermentation broth.
[0056] ...
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