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Biocontrol of Pseudomonas and its application

A technology of Pseudomonas and biocontrol strains, applied in the field of biochemistry, can solve the problem of single antibacterial action mechanism, achieve good control effect, convenient application method, and simple preparation process

Active Publication Date: 2022-04-08
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The single action mechanism of bio-control bacteria makes it easier for pathogenic bacteria to develop resistance

Method used

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  • Biocontrol of Pseudomonas and its application
  • Biocontrol of Pseudomonas and its application
  • Biocontrol of Pseudomonas and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The isolation and screening of biocontrol strain GZ9, the specific steps include:

[0029] Step 1) Isolation and Preservation of Soil Pseudomonas

[0030] The biocontrol strain GZ9 was isolated from the corn rhizosphere soil in Bijie City, Guizhou Province. The isolation process was as follows: Weigh 10 g of the soil sample into a sterile Erlenmeyer flask, add 90 mL of sterile water, and sterilize 5 g of small steel balls at 28 ° C. , 180rpm shaker for 20min, and then stand at room temperature for 5min to obtain 10 -1 Diluent, absorb 100μL and add to 900μL sterile water, mix well to get 10 -2 Diluent, adopt the same method to carry out gradient dilution, obtain 10 -3 and 10 -4 Diluent. 100 μL of each gradient was evenly spread on the Pseudomonas selective medium, and 3 plates were plated for each gradient, placed in a 28°C incubator, and cultured upside down.

[0031] After 1 to 3 days, pick a single colony and grow on LB solid medium. After 1 day, select the strai...

Embodiment 2

[0045] The identification of biocontrol strain GZ9, the specific steps include:

[0046] Step 1) Physiological and biochemical characteristics

[0047] Table 2 The main physiological and biochemical characteristics of strain GZ9

[0048]

[0049]

[0050] Step 2) 16S rRNA gene identification

[0051] Use bacterial 16S rRNA gene universal primer 27 as shown in Seq ID No.4

[0052] F(5'-AGAGTTTGATCCTGGCTCAG-3') and 1492 as shown in Seq ID No.5

[0053] R(5'-TACGGCTACCTTGTTACGACTT-3'), using the genomic DNA of strain GZ9 as a template; carry out PCR reaction according to the following reaction system (30 μL): 2×Taq PCR StarMix with Loading Dye 15 μL, template DNA 1 μL, upstream primer 27F 1.4 μL , downstream primer 1492R 1.4 μL, ddH 2 O 11.2 μL. PCR reaction program: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 90 s, a total of 32 cycles; final extension at 72°C for 10 min. The PCR product is abou...

Embodiment 3

[0056] The preparation of the GZ9 biocontrol agent, the specific steps include: insert the biocontrol strain GZ9 into the LB liquid medium, cultivate it at 28°C and 180rpm for 12h, and make the seed liquid; inoculate the seed liquid with 1% inoculum in the fermentation culture medium, 28°C, 180rpm shaking culture for 48h, to obtain a fermentation broth, the total concentration of viable bacteria was 1×10 9 CFU / mL or more is the prepared biocontrol agent. The fermentation medium still uses LB liquid medium.

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Abstract

A biocontrol Pseudomonas sp. and its application, specifically the biocontrol strain GZ9, with a preservation number of CCTCC M 20191037. The strain has a significant inhibitory effect on Botrytis cinerea, and has good inhibitory effects on Phytophthora capsici, Magnaporthe grisea and Fusarium rubella. The fermented liquid of the strain has a good control effect on tomato gray mold, and the control effect reaches 78.66%. Therefore, the strain has a good application prospect in biocontrol.

Description

technical field [0001] The invention relates to a technique in the field of biochemistry, in particular to a biocontrol pseudomonas and its application. Background technique [0002] Plant diseases are one of the main factors limiting the world's grain production, and the annual grain loss caused by plant diseases can reach 25% of the world's grain production. Among them, 70% to 80% of plant diseases are caused by plant pathogenic fungi. At present, the application of chemical pesticides is the most important way to control plant fungal diseases. However, the long-term and unreasonable use of chemical pesticides has caused a series of problems such as pesticide residues, pathogenic bacteria resistance, and ecological balance damage, which not only has a great impact on food safety and the ecological environment, but also seriously restricts the development of the agricultural industry. [0003] The biological control of plant diseases mainly uses living microorganisms or t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/27A01P3/00C12R1/38
CPCC12N1/20A01N63/00C12R2001/38C12N1/205
Inventor 梁靖聆朱勃聂文翰王沛鸿吴言王赛阿衣则克然木·依明
Owner SHANGHAI JIAO TONG UNIV
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