Corn endophyte Bacillus subtilis and its application in biocontrol
A technology of bacillus and subtilis, applied in the direction of application, bacteria, biocide, etc., can solve the problem of unstable control effect, and achieve the effect of increasing the vitality of the root system
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Embodiment 1
[0016] Embodiment 1: Isolation and identification of Bacillus subtilis BS0521
[0017] Bacterial strain Bacillus subtilis BS0521 of the present invention is isolated from sweet corn seed, and specific operation mode is as follows:
[0018] (1) Sample treatment: Prepare Zhefengtian No. 2 corn seeds, rinse with clean water, and soak in sterile water at 30°C for 30 minutes;
[0019] (2) Surface sterilization: after rinsing with sterile water, place in 95% alcohol for 3 minutes, rinse with sterile water for 4 times immediately after disinfection, and take 120 μL of the last rinse water and apply it to NA medium (beef extract 1g, peptone 5g, yeast extract 5g, sodium chloride 5g, sucrose 10g, agar 20g, water 1000mL), culture at 28°C for 72h, and test the surface sterilization effect;
[0020] (3) Separation: Use sterilized scissors to cut the sterile material on the surface into small pieces, transfer it to a centrifuge tube filled with 1mL sterile water, shake it fully, absorb 0.2...
Embodiment 2
[0028] Example 2: Effect of Bacillus subtilis BS0521 on corn root activity and biomass
[0029] The BS0521 strain was inoculated on the NA medium by the three-section line method, and a single colony was picked and inoculated in the NB liquid medium, cultured at 30°C with shaking at 170r / min, overnight, and ready for use. The BS0521 bacterial suspension was centrifuged at 10 000 r / min for 20 min, the supernatant was carefully drawn, and sterilized by filtration with a 0.22 μm microporous membrane to obtain a BS0521 sterile filtrate for use.
[0030] Soak corn seeds with BS0521 bacterial suspension and sterile filtrate for 30 minutes, sow the seeds, and soak the seeds in clear water as a control. Each treatment was replicated 5 times with 30 seeds per replicate. The emergence rate was counted 7 days after emergence. Harvest corn plants with roots, wash the soil around the roots of the plants, and absorb the water. Root activity of maize seedlings was determined by the method...
Embodiment 3
[0034] Embodiment 3: broad-spectrum determination of Bacillus subtilis BS0521 antibacterial activity
[0035] The pathogenic target bacteria for the broad-spectrum assay of the antibacterial activity of Bacillus subtilis BS0521 are Pseudomonas grandis, Pseudomonas minor, Stem rot and Sheath blight. These four diseases are the main leaf spot diseases and soil-borne diseases on maize plants at present. The specific operation method is as follows: the pathogenic bacteria are cultured in a straight line in the middle of the PDA medium, and BS0521 is inoculated on both sides of the straight line (such as figure 1 Shown), with only the treatment of inoculating pathogenic bacteria as the control, each treatment was repeated 4 times, the growth and changes of the colonies were observed, and the degree of antagonism was judged by the size of the inhibition zone.
[0036] The results showed that BS0521 had significant inhibitory effect on maize leaf spot, small leaf spot and sheath bli...
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