Bacillus pumillus and application thereof in prevention and control of cereal cyst nematodes
A technology of Bacillus pumilus and cereal cyst nematode, which is applied to a strain of Bacillus pumilus and its application in the prevention and treatment of wheat cereal cyst nematode, can solve the problems of no effective biocontrol agents and other problems, and it is not easy to achieve Drug resistance, low production cost, and human and animal safety effects
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Embodiment 1
[0022] Embodiment 1, the acquisition and identification of Bacillus pumilus B202 bacterial strain
[0023] 1. Obtaining of Bacillus pumilus B202 strain
[0024] Severely diseased wheat and healthy wheat were excavated from plots with serious incidence of cereal cyst nematodes in Henan, Hebei, and Shaanxi provinces, the main wheat producing areas in the central plain of China, and the root soil was preserved. Carefully remove the soil at the root of the wheat plant, randomly cut out new tap roots and capillary roots, place them in 50ml of sterile water containing glass beads, and shake them well. The roots are removed to obtain a soil suspension. The root system taken out was first rinsed with sterile water, then sterilized with 1% sodium hypochlorite for 8 minutes, and then rinsed with sterile water for 4 times to rinse off residual sodium hypochlorite. Mix 2 grams of root with 10ml of sterile water and add a small amount of quartz sand to the sterile water. Grind in a funga...
Embodiment 2
[0031] Embodiment 2, the cultivation of bacillus pumilus B202
[0032] 1. Inoculate a single colony of Bacillus pumilus B202 into NA modified medium (solvent is water, containing 3g / L beef extract, 5g / L peptone, 3g / L glucose; pH7.0), shake at 30°C and 180rpm / min Cultivate for 48 hours to get OD 600nm =4.0 fermentation system suspension.
[0033]2. Centrifuge the suspension of the fermentation system obtained in step 1 at 4000 rpm for 30 minutes, and collect the supernatant and bacterial precipitates respectively.
[0034] 3. Suspend the bacterial cell precipitate obtained in step 2 in sterile water to obtain a spore concentration of 8×10 8 spore suspension per ml.
Embodiment 3
[0035] Example 3, Influence of Bacillus pumilus B202 on the activity of wheat cereal cyst nematode under indoor conditions
[0036] 1. Suspending the second-instar larvae of wheat cereal cyst nematodes with sterile water.
[0037] 2. Group processing is as follows:
[0038] Experimental group: 250 μl of the suspension obtained in Step 1 (containing 100±5 second-instar larvae of wheat cereal cyst nematodes) was mixed with 250 μl of the fermentation system suspension obtained in Example 2, and cultured at room temperature for 24 hours.
[0039] Control group: 250 μl of the suspension obtained in step 1 (containing 100±5 second-instar larvae of wheat cereal cyst nematodes) was mixed with 250 μl of sterile water, and cultured at room temperature for 24 hours.
[0040] 3. Add 1mol / L NaOH aqueous solution to each culture system in step 2 (the surviving larvae will move under alkaline stimulation, and the immobile larvae will be counted as dead larvae), and count the number of secon...
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