Bacillus cereus for preventing and controlling plant nematodes and application thereof
A technology of Bacillus cereus and plants, applied in the fields of application, pest control, nematicide, etc., can solve the problems of severe pollution, high toxicity, and environmental damage
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Embodiment 1
[0046] Example 1. Isolation, purification and identification of bacterial strain MH61-63-03
[0047] 1. Isolation and purification of strain MH61-63-03
[0048] Strain MH 61-63-03 was isolated from soil samples collected in Mohe, Heilongjiang. The specific steps are as follows: take 5g of the soil sample, add it into a conical flask containing 45mL of sterile water, shake and cultivate the conical flask for 30min in a shaker with a rotating speed of 200rpm / min, and prepare 10 -1soil dilution. Stepwise dilution was carried out in the ultra-clean workbench to prepare a series of soil dilutions with corresponding gradients. Use a pipette gun to draw an appropriate amount of soil dilution on the LB plate, then spread it evenly with a spreader, and repeat 3 times for each concentration. Place it in a constant temperature incubator at 28°C for 24 hours, observe the growth of the colony on the plate, pick a single colony on the plate, name it MH 61-63-03 strain, and purify it on a...
Embodiment 2
[0057] The preparation of embodiment 2, MH61-63-03 bacterial agent
[0058] 1. Culture of plate strains
[0059] The MH61-63-03 strain isolated in Example 1 was inoculated on an LB solid medium plate, cultured at 28° C. for 24 hours, and a single colony was picked to obtain a plate strain.
[0060] 2. Liquid fermentation culture
[0061] Inoculate the plate strain obtained in step 1 into a 250mL Erlenmeyer flask containing 100mL of fermentation medium, and ferment and culture at 28°C for 72h at a shaker speed of 160r / min to obtain MH61-63-03 and its metabolites. For the fermentation broth, centrifuge the fermentation broth at 10000rpm / min for 5min to obtain the supernatant, and filter it with an organic filter membrane with a pore size of 0.22μm to obtain the fermentation filtrate, which is named MH61-63-03 bacterial agent.
[0062] The prepared MH61-63-03 bacterial agent was diluted 5 times with sterile water to obtain a 5-fold dilution of the MH61-63-03 bacterial agent.
Embodiment 3
[0063] Embodiment 3, the application of MH61-63-03 bacterial agent in the prevention and treatment of plant nematodes
[0064] 1. Nematicidal activity test of MH61-63-03 bacterial agent on second instar larvae of plant parasitic nematodes
[0065] 1. Obtaining the second instar larvae of the root-knot nematode Meloidogyne incognita
[0066] Take water spinach roots inoculated with second instar larvae of Meloidogyne incognita for 60-90 days, clean the root soil with tap water, pick mature and plump egg masses with tweezers under a magnifying glass and place them in the hatching sieve, and place the hatching sieve containing a large number of egg masses Place in a petri dish, add distilled water to the petri dish until the eggs are slightly submerged, and hatch in a constant temperature incubator at 25°C. In order to ensure the complete rate of hatched nematodes, the second instar larvae hatched on the first day are discarded; The activity of the second-instar larvae after hat...
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