A kind of Bacillus subtilis for preventing and treating Fusarium disease of crops and its application
The technology of Bacillus subtilis and biocontrol agent is applied in the field of development and utilization of microbial germplasm resources to achieve the effects of high-efficiency antagonism, good application prospect and stable control efficiency
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Embodiment 1
[0036] The preparation of the biocontrol agent containing BS101 bacterial strain of embodiment 1
[0037] Acquisition of 1BS101 strain
[0038] The BS101 strain was isolated from wheat ears in Hai'an County, Jiangsu Province. Wheat ears were ground and spread on LB plate after 80°C water bath for 10 minutes; colonies were picked for purification and culture the next day. The surface of the colony of strain BS101 on the culture medium is rough, opaque, shrunken and white (see figure 1 ), able to form spores, mobile, aerobic. The 16S rDNA sequence is shown as SEQ ID No.1, and the alignment result of the sequence in NCBI shows that it has the highest homology with Bacillus subtilis.
[0039] 2 Preparation of biocontrol agents
[0040] (1) Inoculate the BS101 strain into the LB culture medium, culture at 30°C and shake at 180rpm for 12 to 16 hours, then take samples in the ultra-clean workbench to measure the OD value at 600nm. to zero;
[0041] The formula of LB culture med...
Embodiment 2B
[0044] The antagonistic activity of embodiment 2BS101 bacterial strain to Fusarium graminearum
[0045] Determination of plate antagonistic activity of 1BS101 strain against Fusarium graminearum
[0046] Using the confrontation culture method, inoculate Fusarium graminearum PH-1 (model bacteria) stored at 4°C on a PDA plate for activation. The round bacteria block that is beaten into a diameter of 6mm. 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. BS101 was inoculated, and the test took Bacillus subtilis type strain PY79, clear water and the chemical pesticide cyclostrobin as the control group. Cultivate at 25°C, and record the size of the inhibition zone after the mycelia cover the plate.
[0047] According to the size of the inhibition zone, the activity of BS101 against Fusarium graminearum PH-1 and Fus...
Embodiment 3B
[0067] The control activity of embodiment 3BS101 bacterial strain to wheat head blight
[0068] 1 Evaluation of the effect of BS101 strain on controlling wheat head blight in light incubator
[0069] Collect wheat ears at the flowering stage, and spray inoculate 10 8 CFU / mL (containing 0.05% Tween20) of the BS101 strain until the water droplets flow (about 5mL per panicle), and then placed in a light incubator, 25°C for 24h. The next day, Fusarium graminearum PH-1 spore suspension (10 5per mL, containing 0.05% Tween20) spray inoculated to wheat ears (spray until small water droplets are formed, about 4 mL per ear), and then placed in a light incubator at 25°C and 90% humidity to observe the disease.
[0070] Four treatment groups were set up in the experiment: BS101, PY79, cyclostrobin and water control. Each treatment group was inoculated with 15 wheat ears, and after inoculation, it was cultivated until the whole ear of the clear water treatment group was onset (see Fig...
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