Application of biocontrol strain BS102 to preventing crops from being polluted by aflatoxin
A technology of BS102 and aflatoxin, applied in the field of crop disease prevention and control, can solve problems such as poor effect, achieve good control effect, save money, and inhibit growth
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Embodiment 1
[0034] Collection, isolation and identification of embodiment 1 BS102 biocontrol strain
[0035] BS102 strain was isolated from peanut rhizosphere soil in Shaoxing City, Zhejiang Province. Take the soil and put it in sterile water, shake it for 1 hour, absorb the supernatant, put it in a water bath at 80°C for 10 minutes, and spread it on an LB plate; pick colonies for purification and culture the next day.
[0036] The colony surface of strain BS102 on LB medium is rough, opaque, shrunken and white (see figure 1 ), able to form spores, mobile, aerobic.
[0037]The DNA of the strain was extracted, the 16S rDNA sequence was amplified, and sequenced. The 16S rDNA of the strain was shown as SEQ ID No.1, and the sequence comparison in NCBI showed that it had the highest homology with Bacillus subtilis .
[0038] The strain has been preserved in the General Microbiology Center (CGMCC) of China Committee for Microbial Culture Collection (CGMCC), located at No. 3, No. 1, Beichen W...
Embodiment 2
[0039] Embodiment 2 Preparation of biocontrol agent
[0040] 1. Preparation of biocontrol agents
[0041] (1) Inoculate the biocontrol strain BS102 into LB culture medium, culture at 30°C with 180r / min shaking, take samples in the ultra-clean workbench after 12 to 16 hours and measure the OD value at 600nm. Bacteria culture solution to zero;
[0042] (2) When the OD value of the bacterial liquid is between 0.5 and 0.8, it is the seed bacterial liquid, and the seed bacterial liquid is added to a 200L fermenter with a fermentation medium at a volume ratio of 1:100, at 30°C 250r / min shaking culture for 20-24 hours, the pH value of the fermentation broth is maintained at 7.0, the dissolved oxygen is 20%, and the ventilation volume is 5-7m 3 / hour, the tank pressure is 0.05-0.1KPa. After the fermentation broth is out of the tank, the gradient dilution plate is used to detect and adjust the concentration of the bacterial suspension to about 1×10 9 cfu / ml.
[0043] The formula ...
Embodiment 3
[0044] Embodiment 3 BS102 biological control bacterial strain is to the determination of the antagonistic activity of Aspergillus flavus
[0045] 1. Determination of antagonistic activity of BS102 against Aspergillus flavus plate
[0046] (1) Test method
[0047] Using the confrontation culture method, inoculate the Aspergillus flavus HZ17 (a strain producing aflatoxin identified in the previous test, which is resistant to pyrimethanil) stored at 4°C on a PDA plate for activation, and wait until the fungus is overgrown on the plate. Use a sterilized puncher to evenly punch out circular bacterial blocks with a diameter of 6 mm from the outer edge of the colony. Inoculate the mycelium block on the center of the WA plate (WA medium / L: peptone 5g, glucose 10g, gravy extract 3g, sodium chloride 5g, agar 20g, pH=7.0), and place them around 25mm away from the center respectively. BS102 was inoculated, and the test took Bacillus subtilis type strain PY79, clear water and 10 μg / ml py...
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