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Biological agent of bacillus methylotrophicus as well as preparation method and application thereof

A methylotrophic, bacillus technology, applied in the field of plant disease biological control, can solve problems such as affecting rice yield, bacteria are prone to drug resistance, polluting the environment, etc., and achieves good environmental compatibility, good control effect, high biological active effect

Active Publication Date: 2013-04-17
陕西恒田生物农业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Rice bacterial blight is caused by Gram-negative bacteria Xanthomonas oryzaepv.oryzae (Xoo), and is one of the three major diseases of rice 1. It occurs in almost all rice growing areas in the world. Except Xinjiang, Tibet and the northern part of Northeast my country, it occurs to varying degrees. The indica rice production areas in the south of the Yangtze River and the Jianghuai Plain are frequent areas, seriously affecting rice production. , currently mainly rely on chemical pesticide control, but long-term use makes the bacteria prone to drug resistance and seriously pollutes the environment

Method used

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  • Biological agent of bacillus methylotrophicus as well as preparation method and application thereof
  • Biological agent of bacillus methylotrophicus as well as preparation method and application thereof
  • Biological agent of bacillus methylotrophicus as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Isolation and screening of methylotrophic bacillus (Bacillus methylotrophilus) in this application:

[0037] (1), isolation of bacillus:

[0038] Weigh 10g of tobacco rhizosphere soil, pour it into a conical flask filled with 90ml of sterilized water, vibrate at 200r / min for 20min, bathe in 80°C water for 30min, dilute by 10 times, apply and separate.

[0039] (2), Purification of Bacillus: According to the characteristics of the bacteria, a single bacterium was picked in time and dropped on the LB solid medium, and purified by the method of streak separation; the purified bacteria were stained by Gram staining and spore staining, showing that the thalline was Rod-shaped, spore-forming, G + The isolate is bacillus; the LB solid medium formula is: yeast extract 5g, peptone 8g, NaCl 5g, agar 15g, water 1000ml, pH 7.4.

[0040] (3), screening of antagonistic bacillus:

[0041] Use TTC medium to select the active R. solanacearum as indicator bacteria, and ma...

Embodiment 2

[0043] Example 2: Identification of Bacillus methylotrophilus:

[0044] The identification process to the methylotrophic bacillus of embodiment 1 gained is:

[0045] (1) Morphological characteristics: LW-6 colonies on NA solid medium are milky white, the surface of the colonies is rough, and the edges are jagged. The electron microscope shows that LW-6 is rod-shaped, Gram-positive, and has perinatal flagella. The size of a single cell is 0.5-0.6um×2.0-2.5um. The formula of the NA solid medium is: 2-3g of beef extract, 5g of peptone, 2.5-3g of glucose, 16-18g of agar, and add water to 1000ml.

[0046] (2), physiological and biochemical experiment: by the physiological and biochemical characteristic test (as shown in Table 1) to methylotrophic bacillus and in conjunction with the morphological characteristic of this bacterial strain, contrast " Bergey's Bacteria Handbook " (Buchanan & Gibbons, 1984 ) and "Common Bacteria System Identification Handbook" (Dong Xiuzhu and Cai M...

Embodiment 3

[0053] Example 3: Cultivation of Bacillus methylotrophilus

[0054] (1), strain activation:

[0055] Medium: LB solid medium, the formula of the LB solid medium is: yeast extract 5g, peptone 8g, NaCl 5g, agar 20g, water 1000ml, pH 7.4;

[0056] Culture conditions: 30°C, 48h;

[0057] Continuous transfer for 3 generations to ensure the purity of the strain, and then inoculated into the fermentation broth for use.

[0058] (2), preparation of seed solution (shake flask fermentation or eggplant bottle fermentation):

[0059] Culture medium: LB liquid medium, the formula of described LB liquid medium is: yeast extract 5g, peptone 8, NaCl 5g, water 1000ml, pH 7.4 (flask volume 250ml, filling capacity 50ml);

[0060] Culture conditions: 30°C, 48h;

[0061] Inoculation volume: 6-8%.

[0062] (3), seed tank fermentation:

[0063] Culture medium: FJ liquid medium, the formula of said FJ liquid medium is: soybean cake powder 1.5%, sucrose 1%, KH 2 PO 4 0.05%, MgSO 4 0.02%, ...

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Abstract

The invention discloses a novel bacillus methylotrophicus, which has the preservation register number of CGMCC No.6793, wherein the strain is separated from tobacco rhizosphere soil and is obtained by separating and purifying and screening strains. The invention further provides a fermentation method of the bacillus methylotrophicus, a fermentation product prepared by the method, as well as a biological agent based on the fermentation product and a preparation method thereof; the invention further provides a use of the biological agent in controlling a variety of plant diseases. According to the invention, the biological agent is good in environmental compatibility so that the number of effective living spores is more than 5,000,000,000 / g, has high biological activity, and in particular has good control effect on tobacco bacterial wilt, wherein the control effect on tobacco bacterial wilt, soft rot of cabbage, rice bacterial leaf blight, rice bacterial stripe and cotton angular leaf spot is up to 80% or more.

Description

technical field [0001] The invention belongs to the field of biological control of plant diseases, in particular, the use of beneficial microorganisms to carry out biological control of plant diseases, in particular to Bacillus methylotrophicus, and the use of the strain to prepare biological agents and the biological agents to prevent and control plants Disease application. Background technique [0002] The prevalence of plant diseases not only causes crop yield reduction, but also causes great economic losses to human beings, and even brings catastrophic consequences. The harm of plant diseases can also lead to poor quality of agricultural products, unsuitable for use or processing, and even lead to poisoning of humans and animals. In the prevention and control of plant diseases, chemical pesticides have once become people's first choice because of their advantages of low cost, quick effect, and time-saving and labor-saving. However, chemical pesticides are generally non-...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02A01P1/00C12R1/07
Inventor 唐满仓翟枫马玉明安德荣
Owner 陕西恒田生物农业有限公司
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