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Preparation method of Paenibacillus and biological control preparation thereof

A technology for biological control preparations and Paenibacillus, applied in the field of microorganisms, can solve problems such as inability to achieve, and achieve the effects of fast growth, good environmental compatibility and high fermentation density

Active Publication Date: 2019-07-19
LUDONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although some existing biological agents made of microorganisms can achieve the effect of disease prevention to a certain extent, they are far from reaching the ideal state that people want.

Method used

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  • Preparation method of Paenibacillus and biological control preparation thereof
  • Preparation method of Paenibacillus and biological control preparation thereof
  • Preparation method of Paenibacillus and biological control preparation thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: the method for bacterial strain KXJ51 isolation and screening

[0022] Isolation: Bacterial strain KXJ51 was separated by dilution coating method. First, weigh 10g of seabed sediment sample and dissolve it in 90mL of sterile water, shake it well to mix the sample with water, spread it on the LB medium plate after gradient dilution, and then The plate was cultured in an incubator at 30°C for 5 days, and the colonies of different forms on the LB medium were streaked on the LB medium plate, and the obtained strains were purified and stored under separate numbers. The LB medium components are: yeast extract powder 5g / L, peptone 10g / L, NaCl 5g / L, agar 20g / L, pH 7.0, sterilized by high pressure steam at 121°C for 20min.

[0023] Screening: Using the confrontation culture method, inoculate Botrytis cinerea at the midpoint of the PDA medium with a hole puncher with a diameter of 5 mm, inoculate 10 μL of antagonistic bacteria on both sides symmetrically at a distan...

Embodiment 2

[0024] Example 2: Identification of bacterial strain KXJ51

[0025] Strain characteristics: two days after culturing the strain KXJ51 with LB medium, the colonies are flat, smooth and round, milky white or light yellow, without pigment; Gram-positive rod-shaped bacteria. Strain KXJ51 can decompose glucose, fructose and glycerol, but cannot utilize mannose and mannitol. The test results of oxidase and nitrate reduction were positive. For the colony morphology of Paenibacillus tyrfis KXJ51 strain, see figure 1 .

[0026] Strain identification: The length of the 16S rRNA gene sequence obtained by DNA extraction, PCR amplification and sequencing is 1412bp, and the BLAST comparison on GenBank shows that the base similarity between the strain and Paenibacillus tyrfis (KT216503) reaches 99%. The strain KXJ51 was identified as Paenibacillus tyrfis based on biological, physiological and biochemical indicators, and the phylogenetic tree based on 16SrRNA sequence figure 2 .

[0027...

Embodiment 3

[0029] Embodiment 3: the fermentation process of Paenibacillus KXJ51

[0030] Strain activation: Pick the strains and inoculate them on a solid slant medium (glucose 2g / L, yeast extract powder 5g / L, peptone 10g / L, NaCl 5g / L, agar 20g / L, pH 7.0, 121℃ high pressure Steam sterilization for 20 minutes), cultured in a constant temperature incubator at 30°C for 2 days, until a large number of bacterial cells grow on the slope to transfer liquid seeds;

[0031] Preparation of liquid seeds: pick activated strains and inoculate them in seed medium (glucose 10g / L, yeast extract powder 5g / L, peptone 10g / L, KH2PO4 1g / L, MgSO4 0.5g / L, corn steep liquor 10g / L, pH7.0, 121°C high-pressure steam sterilization for 20 minutes), sealed with gauze, and cultured at 30°C, 220r / min shaking table for 28h, a large number of bacteria grew, and liquid seeds were obtained;

[0032] Liquid fermentation: 5% inoculum by volume to fermentation medium (glucose 20g / L, peptone 5g / L, yeast extract powder 2g / L, M...

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Abstract

The invention relates to a preparation method of Paenibacillus and a biological control preparation thereof, belonging to the field of microorganisms. Its taxonomic name is: Paenibacillus tyrfisKXJ51, which has been deposited in the China General Microbial Culture Collection and Management Center with the preservation number: CGMCC No.17155, and the preservation date is January 10, 2019. A method for preparing a biological control formulation by Paenibacillus, including strain activation, liquid seed preparation and liquid fermentation. Application of Paenibacillus tyrfisKXJ51 in the control of plant pathogens. The beneficial effects of the invention are as follows: Paenibacillus strain KXJ51 has fast growth speed, high fermentation density, wide action spectrum, can prevent and control various plant diseases, strong genetic stability, good compatibility with the environment, and long duration of disease prevention.

Description

technical field [0001] The invention relates to a preparation method of Paenibacillus and a biological control preparation thereof, belonging to the field of microorganisms. Background technique [0002] Plant diseases are antagonistic symbiosis between host plants and pathogens, and their occurrence and prevalence are the result of the interaction between host plants and pathogens. Diseases of crops and forest trees often cause serious losses to the national economy and people's lives. In recent years, with the public's general concern about the safety of chemical pesticides in food and the environment, and the prohibition of some dangerous chemical pesticides, people are seeking other safe and effective new ways to control plant diseases, and the use of beneficial microorganisms to control plant diseases has attracted increasing attention . It introduces exogenous antagonistic bacteria that are beneficial to plants into the broken plant natural ecosystem, or promotes the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/02A01P1/00A01P3/00C12R1/01
Inventor 解卫海冯志彬郭元柯邵光远
Owner LUDONG UNIVERSITY
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