A strain of Bacillus pumilus and its application
A technology of Bacillus pumilus and bacteria, which is applied in the application field of microbial technology and achieves the effect of significant antagonism
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Embodiment 1
[0042] Example 1, the acquisition of Bacillus pumilus 154-2
[0043] 1. Soil source
[0044] Soil of East Wheat Field in Wanglaoba Village, Yuanzhou District, Guyuan City, Ningxia
[0045] 2. Screening of strains
[0046] (1) Soil sample collection
[0047] Z-shaped 5-point sampling method for soil sample collection: collect 200g soil samples at each point, mix the soil samples evenly, and put 200g of soil samples into a sterilization bag by quartering method as a soil sample. Three soil samples were collected from each plot as replicates. Record the time, place and type of sampling. The collected soil samples were stored in a refrigerator at 4°C for bacterial isolation.
[0048] (2) Isolation of bacteria
[0049] Plate dilution method: Weigh 10 g of soil sample into a conical flask, add 90 mL of sterile water, then shake at 200 rpm and 28 °C in a shaker, take it out after 20 min and let it stand at room temperature for 10 min to prepare a soil bacteria suspension stock ...
Embodiment 2
[0058] Example 2 Identification of 16S rRNA gene of Bacillus pumilus 154-2
[0059] Extracting the genomic DNA of strain 154-2,
[0060] Using primers: 27F 5'-AGAGTTTGATCCTGGCTCAG-3' and 1492R5'-TACGGCTACCTTGTTACGACTT-3', using the extracted DNA as a template, PCR amplification was performed to obtain the target fragment. The PCR reaction system is:
[0061] Table 1 Taq polymerase chain reaction system
[0062]
[0063]
[0064] The basic conditions of PCR reaction were: pre-denaturation at 94°C for 3 min, denaturation at 94°C for 30s, annealing at 56°C for 30s, extension at 72°C for 90s (1 kb / min), pre-extension at 72°C for 8 min, and storage at 4°C for a total of 30 cycles. After the reaction, the PCR products were tested by 1% agarose gel electrophoresis, and the results were detected and recorded by a gel imager. The gel electrophoresis results of the 16S rRNA gene were as follows: image 3 shown, image 3 1 represents the Marker; 2 represents the product of the 16...
Embodiment 3
[0067] Example 3 Physiological and biochemical identification of Bacillus pumilus 154-2
[0068] The physiological and biochemical characteristics of Bacillus pumilus 154-2 of the present invention are: can secrete β-galactosidase, 3-hydroxybutanone to produce acetomethyl carbinol and gelatinase; cannot hydrolyze arginine, lysine, ornithine Acid, sodium citrate and sodium thiosulfate, etc.; cannot oxidize glucose, mannitol, rhamnose and sucrose, etc.; can utilize 20 carbon sources such as L-arabinose, ribose, D-xylose, fructose and mannose Acid production, see Tables 2 and 3.
[0069] Table 2 Physiological and biochemical characteristics of strain 154-2 – enzyme activity, carbon source oxidation
[0070]
[0071]
[0072] +: positive reaction; -: negative reaction;
[0073] Table 3 Physiological and biochemical characteristics of strain 154-2 - acid production using carbon source
[0074]
[0075]
[0076] +: positive reaction; -: negative reaction; W: weak posi...
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