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Bacillus subtilis for controlling tobacco anthracnose

A technology of Bacillus subtilis and inoculum, which is applied in the field of microorganisms, can solve problems such as affecting tobacco leaf production, unbalanced temperature and humidity, and death, and achieves good development prospects, improves safety, and reduces pesticide residues.

Inactive Publication Date: 2012-03-07
中国烟草总公司黑龙江省公司烟草科学研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It occurs in various tobacco-producing areas in my country. Poor management of seedling cultivation in plastic greenhouses, incomplete disinfection, and low temperature and humidity often cause disease within 3-4 days, resulting in slow growth of tobacco seedlings and even a large number of deaths, resulting in a lack of seedling sources. , affecting later tobacco leaf production
[0003]The current prevention and control methods for this disease are mainly chemical pesticides, but long-term use of chemical pesticides will easily cause the bacteria to develop resistance and the damage will become more rampant; The increase of pesticide residues and environmental pollution. Therefore, the research and development and application of biopesticides compatible with the environment are favorable measures to alleviate and change the "three R" (Residue, Resistance, Resurgence) problems caused by chemical pesticides. An important means for the sustainable and healthy development of the industry

Method used

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  • Bacillus subtilis for controlling tobacco anthracnose
  • Bacillus subtilis for controlling tobacco anthracnose
  • Bacillus subtilis for controlling tobacco anthracnose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Acquisition and identification of Bacillus subtilis MKT-01

[0025] 1. Acquisition and identification of Bacillus subtilis MKT-01

[0026] The Bacillus subtilis MKT-01 of the present invention was taken from the rhizosphere of a tobacco plant in Fanjia Township, Ning'an County, Mudanjiang City, Heilongjiang Province in June 2011, and cultured in a flat medium for 4 days after dilution. Coating in medium, culturing for another 4 days, picking a single colony, streaking it in a flat medium, and culturing and storing on a slope after purification.

[0027] This bacterium is a gram-positive bacterium with round or irregular colonies, rough and opaque surface, and white stains. The surface of LB liquid culture medium forms stained white wrinkles. The size of a single cell is 0.7~0.8 um × 2~3 um, perimeter flagella, can swim, spores are 0.6~0.9 um×1.0~1.5 um, elliptical to column shape, the bacteria are almost under the condition of pH value lower than 5.5 and higher tha...

Embodiment 2

[0031] Example 2 Cultivation of Bacillus subtilis MKT-01

[0032] (1) Slant culture: culture time 2 days, culture temperature 28℃, medium composition (1000 mL): peptone 10g, yeast powder 5g, sodium chloride 10g, agar 20g, pH 7.0, high temperature steam at 121℃ Sterilize for 30 min.

[0033] (2) Seed culture: Pick the activated single colony from the inclined surface in the ultra-clean workbench and insert it into the seed culture medium. Shake the bacteria for 24 hours at 28°C and rotate at 110 rpm to obtain the seed liquid. Seed medium components (1000 mL): peptone 10g, yeast powder 5g, sodium chloride 10g, pH 7.0, high temperature steam sterilization at 121 ℃ for 30 min.

[0034] (3) Fermentation: According to 5% of the inoculum, the above-obtained seed liquid was connected to the fermentation medium, and the culture was shaken at 28°C for 2 days (48 hours), and the rotation speed was 110 rpm to obtain the fermentation broth of Bacillus subtilis, which is the present invention T...

Embodiment 3

[0035] Example 3 Indoor antibacterial test of Bacillus subtilis MKT-01 on tobacco anthracnose

[0036] 1 Test materials

[0037] 1.1 Test medium

[0038] LB liquid medium: peptone 10.0 g, yeast powder 5 g, NaCL 10 g, add water to make the volume to 1000 mL, pH 7.0, and steam sterilize at 121 ℃ for 30 min. Used for culturing the fermentation broth of the bacteria to be tested

[0039] PDA solid medium: Wash 200 g of fresh potatoes, peel them and chop them, add 1000 mL of distilled water, boil for 30 minutes, filter the potatoes with 8 layers of gauze, make up 1000 mL of the filtrate with distilled water, then add 20.0 g glucose, 15.0 g Agar powder, divided into aliquots, sterilized by high-temperature steam at 121 ℃ for 30 min. Used for the preservation of pure bacterial strains.

[0040] 1.2 Test drugs

[0041] Agar powder, glucose, peptone, yeast powder, NaCL, etc.

[0042] 1.3 Instruments

[0043] Sterilization pot, light incubator, full temperature oscillator, 1 / 1000 g electronic ba...

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Abstract

The invention discloses bacillus subtilis for controlling tobacco anthracnose. The bacillus subtilis MKT-01 is a biocontrol bacterial strain with a preservation number of CGMCC (China General Microbiological Culture Collection Center) No.5082. The invention further provides a bacterial agent prepared from the bacillus subtilis MKT-01 and a preparation method of the bacterial agent. The bacillus subtilis MKT-01 disclosed by the invention and the bacterial agent thereof have obvious inhibition effects on the tobacco anthracnose, have a control effect as high as 79.22%, and has a good biopesticide development prospect.

Description

technical field [0001] The invention relates to a strain of bacillus subtilis for preventing and treating tobacco anthracnose, belonging to the field of microorganisms. Background technique [0002] Tobacco anthracnose is one of the main diseases of tobacco, which can occur in various growth stages of tobacco, and the most serious damage is at the seedling stage. It occurs in various tobacco-producing areas in my country. Poor management of seedlings in plastic greenhouses, incomplete disinfection, and low temperature and humidity often cause disease within 3-4 days, resulting in slow growth of tobacco seedlings and even a large number of deaths, resulting in a lack of seedling sources. , affecting the later tobacco production. [0003] The current control method of this disease is mainly based on chemical pesticides, but long-term use of chemical pesticides will easily cause the bacteria to develop resistance and the harm will become more rampant; at the same time, it will ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/02A01P3/00C12R1/125
Inventor 孙剑萍孙宏伟郭兆奎王春艳虞艳芳元野贺国强
Owner 中国烟草总公司黑龙江省公司烟草科学研究所