Tomato gray mold antagonistic strain B-04-glu

A tomato gray mold and gray mold technology, applied in the direction of bacteria, fungicides, biocides, etc., can solve the problems affecting tomato quality, affecting human health, polluting the environment, etc., achieving resistance to pesticides, long duration, strong healing effect

Inactive Publication Date: 2012-06-27
SHANDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, if chemical agents are used continuously, the bacteria will gradually develop drug resistance, and the control effect will decrease year by year. Moreover, chemical pesticides will affect the quality of tomatoes, pollute the environment, and affect human health.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Step 1: Screening a strain of Bacillus cereus that is resistant to Botrytis cinerea, i.e. B-04 bacteria, from soil samples infected with Botrytis cinerea;

[0013] Step 2: Inoculate a single colony of B-04 bacteria into LB medium, and culture in a shaking table at 30°C for 20 hours, then inoculate the resulting bacterial suspension with LB medium at a ratio of 1:10, and then inoculate at 30°C Vigorously oscillate on a shaker and cultivate to mid-logarithmic phase for about 3 hours;

[0014] Step 3: Ice-bath the bacterial solution finally obtained in step 2 for 20 minutes, centrifuge to collect the bacterial cells, wash the collected bacterial cells with pre-cooled ultrapure water once, and wash twice with Hepes with a concentration of 1 mM and a pH of 7.0, Wash twice with Hepes at a concentration of 1 mM, shock buffer containing 10% glycerol and pH 7.0, then suspend in 1.5 ml shock buffer, aliquot into 100 μl tubes, and store in a refrigerator at -70°C;

[0015] Step 4...

Embodiment 2

[0017] Its step 1 is with the step 1 of embodiment 1;

[0018] Step 2: Inoculate a single colony of B-04 bacteria into LB medium, and culture in a shaking table at 28°C for 15 hours, then inoculate the resulting bacterial suspension and LB medium at a ratio of 1:10, and then inoculate at 28°C Vigorously oscillate on a shaker and cultivate to mid-logarithmic phase for about 2.5 hours;

[0019] Step 3: Ice-bath the bacterial solution finally obtained in step 2 for 30 minutes, centrifuge to collect the bacterial cells, wash the collected bacterial cells with pre-cooled ultrapure water twice, and wash once with Hepes with a concentration of 1 mM and a pH of 7.0, Wash twice with Hepes at a concentration of 1 mM, shock buffer containing 10% glycerol and pH 7.0, then suspend in 1.5 ml shock buffer, aliquot into 100 μl tubes, and store in a refrigerator at -70°C;

[0020] Step 4: Take a tube of bacteria solution and put it on ice to dissolve during electroshock transformation. After ...

Embodiment 3

[0022] Its step 1 is with the step 1 of embodiment 1;

[0023] Step 2: Inoculate a single colony of B-04 bacteria into LB medium, and culture in a shaking table at 26°C for 10 hours, then inoculate the resulting bacterial suspension and LB medium at a ratio of 1:10, and then inoculate at 26°C Vigorously oscillate on a shaker and cultivate to mid-logarithmic phase for about 2 hours;

[0024] Step 3: Ice-bath the bacteria solution finally obtained in step 2 for 40 minutes, centrifuge to collect the bacteria, wash the collected bacteria twice with pre-cooled ultrapure water, and wash twice with Hepes with a concentration of 1 mM and a pH of 7.0, Wash once with Hepes at a concentration of 1 mM, shock buffer containing 10% glycerol and pH 7.0, then suspend in 2.0 ml shock buffer, aliquot into 100 μl tubes, and store in a refrigerator at -70°C;

[0025] Step 4: Take a tube of bacteria liquid and put it on ice to dissolve during electroshock transformation. After dissolving, add 10 ...

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PUM

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Abstract

The invention provides a tomato gray mold antagonistic strain B-04-glu which is characterized by comprising four steps for the preparation: firstly, a strain of Bacillus cereus for high-efficient antagonistic gray mold is filtered from a soil sample infected by gray mold, and is named as B-04; secondly, bacteria solution which contains the B-04 bacteria is cultured; thirdly, ultrapure water, 1mM Hepes and electric shock buffer solution are used for washing in turn; fourthly, beta-1, 3-dextranase genes are transformed into the B-04 bacteria through electric shock method, and then the antagonistic strain B-04-glu is obtained. The antagonistic strain B-04-glu belongs to biological prevention and cure and does not contain compositions such as pyrilamine, pyrrole, acylamide and so on of chemical pesticide, thereby the antagonistic strain B-04-glu has the advantages of innocuity, no environmental pollution, difficult production of drug resistance, no residue, easy production, long effective period, strong curing ability, good application effect, safety on non-targeted organisms and so on and has wide application prospect.

Description

Technical field [0001] The invention relates to a bacterial strain, in particular to a tomato gray mold antagonistic strain B-04-glu. technical background [0002] Botrytis cinerea Pers is an important worldwide disease caused by the fungus infection of Botrytis cinerea Pers. Not only is the damage to tomato more and more serious, but it has become the main disease of vegetables such as tomato, cucumber, eggplant, zucchini, kidney bean, lettuce, lettuce, and leek. At present, the control of tomato gray mold in production mainly depends on spraying chemical Pesticides, new chemical pesticides with high activity mainly include pyrimidine amines, pyrroles and amides. However, if chemical agents are used continuously, the bacteria will gradually develop drug resistance, and the control effect will decrease year by year. Moreover, chemical pesticides will affect the quality of tomatoes, pollute the environment, and affect human health. Contents of the invention [0003] The o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12N15/56A01N63/02A01P3/00C12R1/085
Inventor 马汇泉李桂霞
Owner SHANDONG UNIV OF TECH
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