Bacillus siamensis and application of the same in controlling tomato root-knot nematode disease
A technology of tomato root-knot nematode and Bacillus, applied in the application, nematicide, bacteria and other directions, can solve the problems of high multiple cropping index, slow action, complex soil environment, etc., to achieve strong stress resistance, robust growth, and novel strains Effect
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Embodiment 1
[0022] Embodiment 1: Mutation screening and identification of Bacillus siamese strain LK0006
[0023] 1. Mutagenesis and screening of strains
[0024] (1) Isolation of starting strain LK0000
[0025] Isolation of Bacillus: Soil sample collection, in the tomato planting greenhouse plot of Baojia Village, Xinjia Street, Longkou City, Shandong Province, using the 5-point sampling method to collect the appropriate amount of soil around and in the center of the plot within a depth of 10-20cm. Mix in equal amounts. Indicate the location, time and person of the collection. Weigh 1g of soil sample into 100mL of sterile water, place it in a shaker at 30°C at 150rpm for 10min, then place it in a water bath at 60°C for 30min, take 100μL of 10 -2 、10 -3 、10 -4 The diluted solution is spread on the LB medium plate, and each gradient is coated in three parallels. After culturing at 30°C for 2 days, the microbial colonies of different forms on the LB medium are picked and streaked on th...
Embodiment 2
[0047] Embodiment 2: preparation fermented liquid
[0048] Among them, the formula of the seed liquid medium: in terms of mass percentage, bean cake powder 3.00%, corn flour 4.00%, glucose 0.40%, fish meal 0.50%, potassium dihydrogen phosphate 0.03%, light calcium carbonate 1.20%, ammonium sulfate 0.30%, chloride Sodium chloride 0.10%, the rest is water, and the pH is adjusted to 7.0-7.2.
[0049] The mass fermentation medium is the same as the seed liquid medium.
[0050] (1) Prepare the seed liquid of Bacillus siamese LK0006: pick a small amount of bacteria from the inclined plane of the test tube of Bacillus siamese LK0006, transfer to the seed liquid culture medium, shake and culture at 28° C. for 24 hours, and this is the seed liquid;
[0051] (2) Inoculate the seed solution prepared in step (1) into a fermenter containing a large amount of sterilized fermentation medium in a sterile environment, cultivate at a constant temperature of 28-30° C. for 36 hours, and the tran...
Embodiment 3
[0052] Example 3: Verification of indoor effect of Bacillus siamese LK0006 fermentation broth on tomato root-knot nematode
[0053] 1. Experimental method
[0054] 1.1 Preparation of root-knot nematode second-instar larvae suspension
[0055] Fresh egg masses of root-knot nematodes were picked from the tomato roots, incubated in a Beman funnel, and collected once after 24 hours for a total of 7 days. The nematode solution was diluted to contain 100 second-instar larvae / mL, and stored at 4°C for later use.
[0056] 1.2 Preparation of egg suspension
[0057] Cut the tomato roots with root-knot nematodes into pieces and put them into a conical flask, add 100mL of 0.5% NaClO solution, shake vigorously for 1-2 minutes, quickly pour them into a 200-500-mesh sieve group and wash them with tap water for 10 minutes, and collect them through a 500-mesh sieve Eggs in the net were diluted to an egg suspension with a concentration of 3000 grains / mL.
[0058] 1.3 Indoor and in vitro mea...
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