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Microorganism-derived agricultural bacteriostatic agent and preparation method thereof

A bacteriostatic agent and microorganism technology, applied in the field of agricultural bacteriostatic agents and their preparation, can solve the problems of narrow bacteriostatic spectrum of chemical bacteriostatic agents, drug resistance of plant pathogens, incomplete degradation, etc. Good bacteriostatic effect and strong cytolytic activity

Active Publication Date: 2021-01-12
QUFU NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The object of the present invention is to provide a kind of microbial source agricultural bacteriostatic agent and preparation method thereof, help to solve the problem that the chemical bacteriostatic agent bacteriostatic spectrum is narrow in the prevention and control of agricultural pathogenic bacteria, it is easy to cause plant pathogenic bacteria to produce drug resistance, degradation is not complete, easy Problems such as residues in the environment, the agricultural antibacterial agent of the present invention is derived from microorganisms, has good antibacterial effect, and has a wide antibacterial spectrum, and can be applied to the prevention and control of agricultural diseases caused by plant pathogenic bacteria

Method used

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  • Microorganism-derived agricultural bacteriostatic agent and preparation method thereof
  • Microorganism-derived agricultural bacteriostatic agent and preparation method thereof
  • Microorganism-derived agricultural bacteriostatic agent and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Extraction and separation of single rhamnolipid components from fermentation broth of conventional rhamnolipid-producing bacteria

[0023] Rhamnolipids synthesized by conventional rhamnolipid-producing bacteria are mixtures of mono-rhamnolipids and di-rhamnolipids. In the embodiment, Pseudomonas aeruginosa ( Pseudomonas aeruginosa ) SG(Zhao et al. Comparative studies on the structural composition, surface / interfaceactivity and application potential of rhamnolipids produced by Pseudomonas aeruginosa using hydrophobic or hydrophobic substrates. BioresourceTechnology, 2020, 295, 122269) as the starting strain.

[0024] LB medium: peptone 10g, yeast powder 5g, NaCl 10g, add deionized water to 1L, pH 7.0.

[0025] Fermentation medium: glycerol 45g, NaNO 3 3.5g, K 2 HPO 4 ∙3H 2 O 4.0g, KH 2 PO 4 3.0g, MgSO 4 ∙7H 2 O1.0g, add deionized water to 1L, pH 7.0.

[0026] Weigh the above reagents with an electronic balance to prepare the corresponding med...

Embodiment 2

[0033] Embodiment 2: Preparation of single rhamnolipid fermentation broth

[0034] Select the rhamnolipid product as a genetically engineered bacterium Pseudomonas stutzeri that is completely mono-rhamnolipid Pseudomonas stutzeri Rhl (Zhao et al. Heterologous production of Pseudomonas aeruginosa rhamnolipid under anaerobic conditions for microbial enhanced oil recovery. Journal of Applied Microbiology 2015, 118: 379-389) as the starting strain for the production of monorhamnolipid.

[0035]Utilize the LB medium and fermentation medium among the embodiment 1, get the slant 1 inoculation loop of bacterial strain Rh1, inoculate in the Erlenmeyer flask containing 100ml LB medium, at 30 ℃, under 180 rpm / min, cultivate 6-10 hours , to OD 600 The value is 0.6~1.0, and the seed solution of strain Rhl is obtained. Then, inoculate the fermentation medium with a 3% inoculum amount (v / v), and culture it for 5 days at 37°C and 180 rpm to ferment and produce monorhamnolipids to obta...

Embodiment 3

[0037] Example 3: Extracting mono-rhamnolipid from fermentation broth of mono-rhamnolipid-producing bacteria

[0038] Centrifuge the fermented liquid obtained from the above-mentioned Example 2 at 8000 g to remove insoluble matter such as bacteria, and use the chloroform / methanol (v / v, 2:1) extraction method in Example 1 to extract the single mouse in the fermented liquid Li glycolipid.

[0039] The extracted monorhamnolipid product was qualitatively verified by the thin-layer chromatography method in Example 1.

[0040] The extracted mono-rhamnolipid is used to prepare an aqueous solution, wherein the concentration of the mono-rhamnolipid is 200-2000 mg / L, which is used as a bacteriostatic agent for plant pathogenic bacteria.

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Abstract

The invention relates to the field of prevention and treatment of agricultural pathogenic bacteria, in particular to a microorganism-derived agricultural bacteriostatic agent and a preparation methodthereof. The active component of the agricultural bacteriostatic agent is monorhamnolipid produced by microbial fermentation, and the concentration of monorhamnolipid in the agricultural bacteriostatic agent is 0.01 wt%-0.5 wt%. The invention also provides a preparation method of the agricultural bacteriostatic agent. The preparation method comprises the following steps: (1) strain fermentation ofmonorhamnolipid producing bacteria or conventional rhamnolipid producing bacteria; (2) pretreatment of monorhamnolipid producing bacterium fermentation liquor, or separation of a monorhamnolipid component from monorhamnolipid producing bacterium fermentation liquor extract; and (3) dilution of the monorhamnolipid producing bacterium fermentation liquor, or preparation of a monorhamnolipid solution so as to prepare the bacteriostatic agent. The agricultural bacteriostatic agent disclosed by the invention is a microorganism source, has a good bacteriostatic effect and a wide bacteriostatic spectrum, and can be applied to prevention and treatment of agricultural diseases caused by phytopathogens.

Description

technical field [0001] The invention relates to the field of agricultural biocontrol, in particular to an agricultural antibacterial agent derived from microorganisms and a preparation method thereof. Background technique [0002] Phytopathogenic bacteria can cause crops to wilt, rot, spots, white leaves, blight, blight, etc., and even cause plant death. Plant pathogenic bacteria not only threaten the yield of crops, but also affect the quality and safety of agricultural products. Plant diseases caused by plant pathogens have brought huge economic losses to agriculture, which makes the control of agricultural pathogens have important theoretical research and practical application value. The emergence of chemical antibacterial agents has brought effective control methods for the harm of agricultural pathogens. Chemical antibacterial agents often have defects such as narrow antibacterial spectrum and easy to cause plant pathogenic bacteria to develop drug resistance. At the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N43/16A01P1/00C07H15/04C07H1/08C12P19/44C12R1/38
CPCA01N43/16C07H15/04C07H1/08C12P19/44
Inventor 赵峰董梅
Owner QUFU NORMAL UNIV
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