A Strain of Bacillus Velez 229-15 and Its Application
A technology of Bacillus and Veles, applied in the field of microorganisms, can solve the problems of lack of disease-resistant varieties, ecological environment damage, etc., and achieve good antagonistic effect and broad-spectrum antibacterial ability
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Embodiment 1
[0049] Example 1, Isolation, screening and purification of Bacillus veleisi 229-15
[0050] 1. Source of the strain
[0051] The pathogenic strains used are all strains preserved by our laboratory, and all of them are published strains.
[0052] 2. Acquisition of antagonistic strains
[0053] (1) Discovery of antagonistic strains
[0054] In the laboratory, a strain capable of inhibiting the growth of bacterial blight of rice was screened from campus soil samples collected by Gansu Agricultural University in Anning District, Gansu Province, China.
[0055] (2) Purification and preservation of strains
[0056] Using plate streaking method: Pick out the colony that can inhibit the growth of bacterial blight, streak on a new NA plate, place it in a biochemical incubator at 28°C, and culture it upside down for more than 12 hours, pick a single colony, and number it for 229-15. Inoculate the strain 229-15 in the NA liquid medium, culture it in a shaker at 28°C and 180rpm for 1...
Embodiment 2
[0059] 16S rRNA gene identification of embodiment 2, bacterial strain 229-15
[0060] Use bacterial universal primer 27F 5'-AGAGTT TGA TCCTGGCTCAG-3' and 1492R 5'-TACGGCTACCTTGTTACGACTT-3', and using the genomic DNA of strain 229-15 as a template; prepare a 30 μL PCR reaction system according to the ratio in Table 1, and perform the PCR reaction:
[0061] Table 1 Taq polymerase chain reaction system
[0062]
[0063] The basic conditions of the PCR reaction were set as follows: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 30 s, annealing at 56°C for 30 s, extension at 72°C for 90 s, a total of 30 cycles; pre-extension at 72°C for 10 min. After the PCR reaction, the PCR product amplified by strain 229-15 was tested by 1% agarose gel electrophoresis, and the image was detected by a gel imager, and then the original solution of the PCR product was sent to Platinum Biotechnology (Shanghai) Co., Ltd. sequencing.
[0064]The obtained sequencing results were an...
Embodiment 3
[0066] Embodiment 3, the morphology and physiological and biochemical identification of bacterial strain 229-15
[0067] Bacterial strain 229-15 morphological observation of the present invention, such as figure 1 As shown, the colony of bacterial strain 229-15 was found to be milky white, with rough and irregular edges, dry and rough surface, and opaque; the result of Gram staining was positive. Physiological and biochemical test results of bacterial strain SA001 of the present invention show that the table bacterial strain has enzymatic activity to pyruvate and Kohn gelatin; but it cannot oxidize glucose, mannitol, inositol, sorbitol, rhamnose, sucrose, honey Sugar, amygdalin and arabinose; 17 kinds of carbon sources such as glycerin, L-arabinose and D-xylose can be used to generate acid. (See Table 2 and Table 3).
[0068] Table 2 Physiological and biochemical characteristics of strain 229-15 - enzyme activity, carbon source oxidation
[0069]
[0070] +: Positive rea...
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