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
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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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