Bacillus velezensis, separation method and application thereof

A technology of Bacillus Velez and Bacillus, applied in the application field of microbial technology, can solve problems such as polluting the environment, difficult large-scale promotion of agricultural control, endangering human and animal health, and achieves a significant antagonistic effect

Active Publication Date: 2019-02-22
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chemical control has the disadvantages of polluting the environment, endangering the health of humans and animals, and easily causing pathogenic bacteria to mutate and produce drug resistance; agricultural control is difficult to promote on a large scale

Method used

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  • Bacillus velezensis, separation method and application thereof
  • Bacillus velezensis, separation method and application thereof
  • Bacillus velezensis, separation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The acquisition of embodiment 1, Veles bacillus 504

[0037] 1. Soil source

[0038] Soil of water spinach fields in Youxi County, Sanming City, Fujian Province

[0039] 2. Screening of bacterial strains

[0040] (1) Soil sample collection

[0041] Zig-shaped 5-point sampling method for soil sample collection: collect 200g soil sample at each point, mix the soil sample evenly, take 200g by quartering method and put it into a sterilization bag 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 use in bacterial isolation.

[0042] (2) Isolation of bacteria

[0043] Plate dilution method: Weigh 10g of soil sample into an Erlenmeyer flask, add 90mL of sterile water, then shake at 200rpm in a shaker at 28°C, take it out after 15min and let it stand at room temperature for 5min to make a stock solution of soil bacteria suspensio...

Embodiment 2

[0051] The 16S rRNA gene identification of embodiment 2 Veles bacillus 504

[0052] Genomic DNA of strain 504 was extracted using primer: 27F

[0053] 5'-AGAGTT TGA TCCTGGCTCAG-3' and 1492R

[0054] 5'-TACGGCTACCTTGTTACGACTT-3', using the extracted gDNA as a template, perform PCR amplification to obtain the target fragment. The PCR reaction system is:

[0055] Table 1 Taq polymerase chain reaction system

[0056]

[0057] The basic conditions of the PCR reaction were: pre-denaturation at 94°C for 3 min, denaturation at 94°C for 30 s, annealing at 56°C for 30 s, extension at 72°C for 90 s (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 checked by 1% agarose gel electrophoresis, and the gel imager was used to detect and record the results (see attached Figure 4 ). Send the PCR stock solution to Bosun Biotechnology (Shanghai) Co., Ltd. for sequencing. The sequencing results were analyz...

Embodiment 3

[0059] The gyrA gene identification of embodiment 3 Veles bacillus 504

[0060] Genomic DNA of strain 504 was extracted, using primers: GyrA-F 5'-CAGTCAGGAAATGCGTACGTCCTT-3' and GyrA-R 5'-CAAGGTAATGCTCCAGGCATTGCT-3', using the extracted DNA as a template, PCR amplification was performed to obtain the target fragment. The PCR reaction system is:

[0061] Table 2 Taq polymerase chain reaction system

[0062]

[0063] Reaction conditions: 94°C 10min; 94°C 1min, 55°C 1min, 72°C 1min, 30 cycles; 72°C 10min, 10°C∞. After the reaction, the PCR products were checked by 1% agarose gel electrophoresis, and the gel imager was used to detect and record the results (see attached Figure 5 ). Send the PCR stock solution to Bosun Biotechnology (Shanghai) Co., Ltd. for sequencing. The sequencing results were analyzed by DNAStar and compared with BLAST on the NCBI website to determine the species of closely related bacteria.

[0064] The results showed that the gyrA gene of strain 504 ...

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Abstract

The invention relates to a bio-control bacillus velezensis, a separation method and an application thereof. The bacillus velezensis has been preserved in China Center for Type Culture Collection on July 25, 2018; the preserving name is bacillus 504 or bacillus velezensis 504; the preserving number is CCTCC NO: M2018493. The bacillus velezensis provided by the invention has an obvious inhibiting effect to xanthomonas oryzae pv, has broad-spectrum bacteriostatic activity to xanthomonas oryzae pv. oryzicola, banana bacterial wilt, campestris pv.vesicatoria, walnut bacterial black spot, cowpea fusarium wilt, and the like, and has a significant bio-control application prospect.

Description

technical field [0001] The invention belongs to the field of microbial technology application, and in particular relates to a strain of Bacillus velesii and its isolation method and application. Background technique [0002] Rice bacterial blight (Bacterial Blight, BB) and rice bacterial leaf spot (Bacterial Leaf Streak, BLS) are common and important bacterial diseases in rice production, which are caused by Xanthomonasoryzae pv. and X.oryzae pv.oryzicola, Xoc (Zou Lifang, Zhou Dan, Liu Zhiyang, Zou Huasong, Chen Gongyou. Establishment of an insertion mutation system for Xanthomonas oryzae pathogenicity-related genes[J]. Phytopathological Journal , 2012, 42(02):176-185.). The infection and reproduction of Xanthomonas mainly depend on the toxic factors such as polysaccharide, adsorbin, type III secretion system and lipopolysaccharide secreted in the host tissue (Tang Qinghua, Zhang Shiqing, Niu Xiaoqing, Zhu Hui, Yu Fengyu, Qin Weiquan. Xanthomonas Research progress on regu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02A01N63/00A01P1/00C12R1/07
CPCA01N63/00C12N1/02C12N1/20C12N1/205C12R2001/07
Inventor 陈颖邹丽芳杨瑞环李生樟刘昭张翠萍陈功友
Owner SHANGHAI JIAO TONG UNIV
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