Biocontrol pseudomonas sp. and application thereof

A technology of Pseudomonas and bio-control strains, applied in the field of biochemistry, can solve the problem of single mechanism of anti-bacteria, achieve good control effect, simple preparation process, and short time-consuming effect

Active Publication Date: 2020-05-22
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 pseudomonas sp. and application thereof
  • Biocontrol pseudomonas sp. and application thereof
  • Biocontrol pseudomonas sp. and application thereof

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 a strain ...

Embodiment 2

[0044] The identification of biocontrol strain GZ9, the specific steps include:

[0045] Step 1) Physiological and biochemical characteristics

[0046] Table 2 The main physiological and biochemical characteristics of strain GZ9

[0047]

[0048]

[0049] Step 2) 16S rRNA gene identification

[0050] Use bacterial 16S rRNA gene universal primer 27 as shown in Seq ID No.4

[0051] F(5'-AGAGTTTGATCCTGGCTCAG-3') and 1492 as shown in Seq ID No.5

[0052] R(5'-TACGGCTACCTTGTTACGACTT-3'), using the genomic DNA of strain GZ9 as a template; carry out PCR reaction according to the following reaction system (30μL): 2×Taq PCR StarMix with Loading Dye 15μL, template DNA 1μL, upstream primer 27F 1.4μL , 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, and extension at 72°C for 90 s, a total of 32 cycles; finally, extension at 72°C for 10 min. The PCR product is a...

Embodiment 3

[0055] 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

The invention relates to a biocontrol pseudomonas sp. and an application thereof, specifically to a biocontrol strain GZ9 with a preservation number of CCTCC M 20191037. The strain has a significant inhibition effect on botrytis cinerea, and shows a good inhibition effect on phytophthora capsici, magnaporthe grisea and gibberella zeae. The fermentation liquor of the strain has a good control effect on tomato gray mold, and can reach a control effect of 78.66%. Thus, the strain has good biocontrol application prospect.

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 irrational 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 the...

Claims

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Application Information

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Patent Type & Authority Applications(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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