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Biocontrol Bacillus velezensis SF259 and application thereof

A technology of Bacillus, Veles, applied in the field of microbial technology application

Active Publication Date: 2020-05-15
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, there are certain limitations in the prevention and control of these two diseases.

Method used

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  • Biocontrol Bacillus velezensis SF259 and application thereof
  • Biocontrol Bacillus velezensis SF259 and application thereof
  • Biocontrol Bacillus velezensis SF259 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The acquisition of embodiment 1, Veles bacillus SF259

[0040] 1. Soil source

[0041] Soil of cherry trees in the orchard of the horticultural field in Ledu District, Qinghai Province, China

[0042] 2. Screening of bacterial strains

[0043] (1) Soil sample collection

[0044] 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.

[0045] (2) Isolation of bacteria

[0046] Plate dilution method: Weigh 10g of soil sample into a Erlenmeyer flask, add 90mL of sterile water, then shake at 200rpm, 28°C shaker, take it out after 20min and let it stand at room temperature for 10min to make the stock solution ...

Embodiment 2

[0053] Example 2 Identification of the 16S rRNA gene of Bacillus Velez SF259

[0054] Genomic DNA of strain SF259 was extracted using primers:

[0055] 27F 5'-AGAGTTTGATCCTGGCTCAG-3', and

[0056] 1492R 5'-TACGGCTACCTTGTTACGACTT-3', with the extracted DNA as a template, PCR amplification was performed to obtain the target fragment. The PCR reaction system is:

[0057] Table 1 Taq polymerase chain reaction system

[0058]

[0059] 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 ...

Embodiment 3

[0062] Example 3 Identification of the gyrA gene of Bacillus Velez SF259

[0063] The genomic DNA of the strain SF259 was extracted, and the primers: GyrA-F 5'-CAGTCAGGAAATGCGTACGTCCTT-3' and GyrA-R 5'-CAAGGTAATGCTCCAGGCATTGCT-3' were used to perform PCR amplification using the extracted DNA as a template to obtain the target fragment. The PCR reaction system is:

[0064] Table 2 Taq polymerase chain reaction system

[0065]

[0066]

[0067] 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, detected by a gel imager, and the results were recorded. 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.

[0068]The results showed that the similarity between the gy...

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Abstract

The invention relates to biocontrol Bacillus velezensis SF259 and application thereof. The Bacillus velezensis strain is collected in the China Center for Type Culture Collection (CCTCC) on September20, 2019, with the collection name of Bacillus velezensis SF259 and the collection number of CCTCCM 2019735. The Bacillus velezensis has a significant inhibition effect on xanthomonas oryzae pv. oryzae (Xoo), also shows broad-spectrum bacteriostatic activity for xanthomonas oryzae pv. oryzicola (Xoc), X. campestris pv. musacearum, X. axonopodis pv. vignicola, X. campestris pv. malvacearum, botrytis cinerea and magnaporthe oryzae, and has important biocontrol application prospects.

Description

technical field [0001] The invention belongs to the field of microbial technology application, and in particular relates to a strain of Bacillus Velez for biocontrol and its application. Background technique [0002] Rice is an extremely important food crop in the world, especially in Asia. Two pathogenic strains of Xanthomonas oryzae, X.oryaze pv.oryaze (Xoo) and X.oryaze pv.oryzicola (Xoc), can infect rice and cause bacterial leaf blight (BLB) and Rice stripe disease (bacterial leaf streak, BLS). These two diseases seriously endanger the yield and quality of rice, and occur heavily in tropical and subtropical Asian regions. [0003] Rice bacterial blight is one of the three major diseases of rice, and it occurs in all rice regions around the world. In my country, rice bacterial blight is also known as ground fire or white leaf blast. After rice is infected with the disease, the yield will be reduced by 20 %-30%, up to 50%-60% in severe cases (Ou, S, H., Rice diseases. Co...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/22A01P1/00A01P3/00C12R1/07
CPCA01N63/00C12N1/205C12R2001/07
Inventor 邹丽芳黄梦桑李生樟杨瑞环陈路生屈伊凝陈功友
Owner SHANGHAI JIAO TONG UNIV
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