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A strain of Bacillus subtilis that is also resistant to tobacco wildfire disease and angular leaf spot

A technology of Bacillus subtilis and bacterial agents, applied in the field of microorganisms, can solve the problems of unstable tobacco field ecosystem, polluted environment, and aggravated disease epidemics, and achieve the effect of good prospects for the development of biological pesticides

Inactive Publication Date: 2011-12-21
中国烟草总公司黑龙江省公司牡丹江烟草科学研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Tobacco wildfire disease and angular leaf spot are the main bacterial diseases of tobacco, which occur in various smoking areas in the world. The current tobacco planting varieties have no resistance to the two diseases, so the prevalence of the diseases is increasing year by year, causing comparative serious loss
[0003] The disease is mainly controlled by chemical pesticides, but the long-term use of chemical pesticides has caused problems such as increased pesticide residues in tobacco leaves and polluted the environment, resulting in The instability and serious deterioration of the disease, the resistance of diseases and insect pests increased, and the damage became more rampant

Method used

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  • A strain of Bacillus subtilis that is also resistant to tobacco wildfire disease and angular leaf spot
  • A strain of Bacillus subtilis that is also resistant to tobacco wildfire disease and angular leaf spot
  • A strain of Bacillus subtilis that is also resistant to tobacco wildfire disease and angular leaf spot

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Acquisition and identification of Bacillus subtilis NYJ-02

[0024] 1. Acquisition and identification of Bacillus subtilis NYJ-02

[0025] The bacillus subtilis NYJ-02 described in the present invention is isolated from tobacco leaves. This bacterium is a Gram-positive bacterium with perinatal flagella and can swim. The size of a single cell is 0.7-0.8 um × 2-3 um, and the spore is 0.6-0.9 um × 1.0-1.5 um. It is elliptical to columnar, and the surface of the colony is rough and opaque. Stained white, stained white wrinkles formed on the surface of the seed culture medium. Under the condition of pH value lower than 4 and higher than 10, bacteria can hardly grow, and the optimum pH value is 7. Optimum medium composition: Nitrogen source (yeast: egg = 5:3) 1%, glucose 0.1%, NaCl 0.3%, KNO 3 0.5%, the optimum growth temperature is 26-28℃.

[0026] The 16S rDNA sequence of the Bacillus subtilis NYJ-02 described in the present invention is shown in SEQ ID No:...

Embodiment 2

[0029] The cultivation of embodiment 2 Bacillus subtilis NYJ-02

[0030] (1) Incline culture: culture time 3 days, medium composition (1000 mL): yeast extract 6.25 g, peptone 3.75 g, glucose 1.0 g, NaCl 3.0 g, KNO 3 5.0 g, agar powder 15.0 g, pH 7.0~7.5. The culture temperature is 26, and the pH is 7.0~7.5. The culture temperature is 26°C~28°C.

[0031] (2) Seed culture: Pick activated single colony from the slope in the ultra-clean workbench and insert it into the seed medium, shake the bacteria at 26°C~28°C for 16~18 hours, and rotate at 200rpm to obtain the seed liquid. Seed medium composition (1000 mL): yeast extract 6.25 g, peptone 3.75 g, glucose 1.0 g, NaCl 3.0 g, KNO 3 5.0 g, pH 7.0~7.5.

[0032] (3) Fermentation: According to the inoculum amount of 5%, the above-mentioned obtained seed liquid was inserted into the fermentation medium, shaken for 3 days (72 hours), and the rotation speed was 200rpm to obtain the Bacillus subtilis fermentation liquid, which is t...

Embodiment 3

[0033] Example 3 Indoor Bacteriostatic Test of Bacillus subtilis NYJ-02 on Tobacco Angular Spot and Wildfire Disease

[0034] Pick the purely cultured pathogenic bacteria (Aerobic spot fungus and wildfire fungus) and Bacillus subtilis NYJ-02 into the Erlenmeyer flasks containing 50 mL of the above seed medium, and place the Erlenmeyer flasks in a constant temperature shaking incubator at 28 °C Shake culture for 48 h, and prepare the concentration of 3 × 10 8 The bacterial suspension of cfu / mL is ready for use.

[0035] Take 5 mL of fermented liquid of Leptosphaeria spp. and Wildfire Bacteria respectively, and add them to 100 mL of the above-mentioned sterilized slant medium that has been melted and cooled to 55°C, shake gently to mix with the medium evenly, and pour into a sterilized petri dish In, cool to a flat plate. Use a pipette gun to draw 70 μL of Bacillus subtilis NYJ-02 fermentation broth and streak it on the cooled plate. After culturing at 28°C for 48-72 h, obs...

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PUM

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Abstract

The invention discloses bacillus subtilis capable of preventing tobacco wildfire and angular leaf spot. The collection number of the bacillus subtilis NYJ-02 is CGMCC No.4555. The invention also provides a bacterial agent prepared by the bacillus subtilis NYJ-02 and a preparation method thereof. The bacillus subtilis NYJ-02 is biocontrol bacteria, the strain and the fungicide of the bacillus subtilis have stronger restraint function to the tobacco wildfire and the angular leaf spot, and thus, the bacillus subtilis has a good biopesticide development prospect.

Description

technical field [0001] The invention relates to a strain of Bacillus subtilis capable of preventing tobacco wildfire disease and angular leaf spot, belonging to the field of microorganisms. Background technique [0002] Tobacco is one of the important economic crops in our country. Tobacco wildfire disease and angular leaf spot are the main bacterial diseases of tobacco, which occur in various smoking areas in the world. The current tobacco planting varieties have no resistance to the two diseases, so the prevalence of the diseases is increasing year by year, causing comparative serious loss. [0003] The prevention and control method of this disease is mainly based on chemical pesticides, but the long-term use of chemical pesticides has caused problems such as increased pesticide residues in tobacco leaves and polluted the environment, leading to instability and serious deterioration of the tobacco field ecosystem, increased resistance of diseases and insect pests, and har...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02A01P1/00C12R1/125
Inventor 孙剑萍孙宏伟郭兆奎虞艳芳
Owner 中国烟草总公司黑龙江省公司牡丹江烟草科学研究所
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