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Bacillus subtilis BBD012 and application to prevent and treat tomato disease

A technology of Bacillus subtilis and BBD012, which is applied in the field of microorganisms, can solve the problems of decreased control effect of chemical agents, endangering human and animal health, and pollution of agricultural products and the environment by chemical agents.

Inactive Publication Date: 2017-09-15
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, long-term and continuous drug use has produced resistance to botrytis cinerea and leaf mold, leading to a decline in the control effect of chemical agents year by year, and chemical agent residues seriously pollute agricultural products and the environment, endangering the health of humans and animals

Method used

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  • Bacillus subtilis BBD012 and application to prevent and treat tomato disease
  • Bacillus subtilis BBD012 and application to prevent and treat tomato disease
  • Bacillus subtilis BBD012 and application to prevent and treat tomato disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Potted tomato control using preparations to prevent tomato gray mold:

[0021] Step 1. Inoculate a single colony of Bacillus subtilis BBD012 in 200mL LB liquid medium, in a constant temperature shaking incubator at 28°C for 160r.min -1 Cultivate for 24 hours, and dilute the cultured bacterial solution to 4×10 8 cfu.mL – 1 , preparing a preparation for preventing tomato gray mold;

[0022] Step 2: Take potted tomatoes that grow to 5-6 leaves and have the same growth, spray the front and back of the tomato leaves with the preparation for treating and preventing tomato gray mold, and use clean water as a control. After 24 hours, the spore suspension of tomato gray mold 1×10 7 spores / mL (Pour a little sterile water into the PDA plate covered with Botrytis cinerea, scrape off the spores on the surface of the agar, pour them into a sterile container, shake them fully, filter them with sterile gauze, and count them on the blood cell plate) spray inoculated on it. Tomato p...

Embodiment 2

[0028] Potted tomato control with preparations for the prevention of tomato leaf mold:

[0029] Step 1. Inoculate a single colony of Bacillus subtilis BBD012 in 200mL LB liquid medium, in a constant temperature shaking incubator at 28°C for 160r.min -1 Cultivate for 24 hours, and dilute the cultured bacterial solution to 4×10 8 cfu.mL – 1 , preparing a preparation for preventing tomato gray mold;

[0030] Step 2: Take potted tomatoes that grow to 5-6 leaves and have the same growth, spray the front and back of the tomato leaves with the preparation for treating and preventing tomato gray mold, and use clean water as a control. After 24 hours, the spore suspension of tomato gray mold 1×10 7 spores / mL (Pour a little sterile water into the PDA plate covered with Botrytis cinerea, scrape off the spores on the surface of the agar, pour them into a sterile container, shake them fully, filter them with sterile gauze, and count them on the blood cell plate count) spray inoculated ...

Embodiment 3

[0036] Screening, isolation and purification of Bacillus subtilis BBD012 strain:

[0037] Step 1, preparation of culture medium:

[0038] LB medium for bacterial plate culture (tryptone 10g / L; yeast extract 5g / L; sodium chloride 10g / L; agar powder 18g / L), LB liquid medium for liquid culture (without adding agar powder), fungi PDA medium (potato 200g / L; glucose 20g / L; agar powder 18g / L) was used for culture and plate confrontation test.

[0039] Step 2: Collect 3g leaves of healthy white lentil plants, soak them in 75% alcohol for 30s, then sterilize them with 1% sodium hypochlorite for 3 minutes, rinse them with sterile distilled water for 5 times, take the last rinse solution 200μL and spread it on LB On the culture medium plate; the sterilized leaves were ground in a sterile mortar, mixed with 5ml of sterile distilled water, and 200 μL of the grinding liquid was taken and spread on the LB medium plate. Cultivate at 28°C and observe day by day. If there is no colony in the ...

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Abstract

The invention discloses Bacillus subtilis BBD012, which is classified and named as Bacillus subtilis; the preservation number is M 2019097. The Bacillus subtilis BBD012 can be used for inhibiting botrytis cinerea and / or cladosporium fulvum cooke. The preparation is Bacillus subtilis BBD012 cultivating fluid. The preparation method includes steps of inoculating single colony of Bacillus subtilis BBD012 to an LB liquid culture medium; and cultivating in a constant-temperature shaking incubator at 28 DEG C for 24 hours at 160 r.min-1; diluting the colony of bacteria solution after being cultured until 4x108 cfu.mL-1 by sterile water; the prevention and treatment method includes step of spraying a tomato plant by the Bacillus subtilis BBD012 culture liquid. The Bacillus subtilis BBD012 has good antagonistic effect to botrytis cinerea and cladosporium fulvum cooke; the prevention and treatment effects to botrytis cinerea and cladosporium fulvum cooke can reach 64.9% and 58.0% respectively; therefore, the Bacillus subtilis BBD012 has very good disease preventing effect.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a bacillus subtilis BBD012 and its application in preventing and treating tomato diseases. Background technique [0002] Botrytis cinerea is a serious and common disease on tomatoes, which occurs in all vegetable areas. In addition to harming tomatoes, it can also harm more than 20 kinds of crops such as eggplant, pepper, cucumber, and gourd. Years with low temperature and continuous rainy weather are serious hazards. When the disease is severe, the stems and leaves die and a large number of rotten flowers and fruits are caused, which directly affects the yield. Tomato gray mold has become the main disease of tomato production in my country's protected areas. Botrytis cinerea is caused by the fungus Botrytis cinerea. [0003] Tomato leaf mold is caused by the fungus Tomato leaf mold ( Fulvia fulva ) infection, symptoms will appear on the leaves, stems, flowers, and fru...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P3/00A01G13/00C12R1/125
CPCA01G13/00A01G2013/004A01N63/00C12N1/20C12N1/205C12R2001/125
Inventor 李永丽周洲崔林开胡艳红闫作炳王少博林乾坤王亚红
Owner HENAN UNIV OF SCI & TECH
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