Biocontrol compound bacterial agent and its preparation method and application
A compound bacterial agent and bio-control technology, applied in biochemical equipment and methods, microbial-based methods, botany equipment and methods, etc., can solve problems such as unsatisfactory control effects, avoid negative impacts, increase income, and promote strong effect
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Embodiment 1
[0026] Embodiment 1: the preparation of biocontrol complex bacterial agent
[0027] This embodiment provides a preferred preparation method of a biocontrol compound bacterial agent. The microbial active components of the biocontrol complex bacterial agent include Streptomyces KN37, Trichoderma harzianum and Bacillus subtilis. Streptomyces KN37 was inoculated into Gao's No. 1 medium, Trichoderma harzianum was inoculated into PDA medium, and Bacillus subtilis was inoculated into LB medium. The culture conditions of the three bacteria were temperature 32°C, rotation speed 180rpm, and shaking culture for 36 hours. Streptomyces KN37 suspension, Trichoderma harzianum suspension, and Bacillus subtilis suspension were obtained respectively. These three kinds of bacterial suspensions were respectively introduced into 50L fermentors equipped with fermentation broth at a mass ratio of 1:100. The formula of the fermentation culture liquid is: millet 10g / L, soybean flour 10g / L, glucose 1...
Embodiment 2
[0028] Embodiment 2: Screening of the optimum ratio of biocontrol compound bacterial agents
[0029]Streptomyces KN37 was used as quantification, Trichoderma harzianum and Bacillus subtilis were used as variables, and the CCD (Central Composite Design) response surface experiment was designed and tested with Design Expert 8.0 software as shown in Table 1. Take 100 mg of Streptomyces KN37 bacterial powder, add corresponding Trichoderma harzianum bacterial powder and Bacillus subtilis bacterial powder respectively according to the experimental design in Table 2, mix well and dissolve in 1L of water, and conduct potted efficacy tests on cotton verticillium wilt respectively.
[0030] Table 1. Selected variables and parameter levels in the CCD experimental design
[0031]
[0032] Table 2, CCD test design scheme and results
[0033]
[0034] Response surface analysis was used to statistically analyze the results, and a multinomial fitting regression model was obtained: Y=64...
Embodiment 3
[0038] Embodiment 3: Cotton verticillium wilt pot experiment
[0039] Take the 5-leaf stage cotton seedlings with consistent growth as the test crop, dilute the biocontrol compound bacterial agent prepared in the embodiment to a certain concentration (0.2, 0.4, 0.6g / L), take 50mL for root irrigation, and irrigate the roots of 10mL cotton after 5 days Verticillium dahliae spore suspension (10 7 CFU / mL), continue to cultivate, irrigate the roots with 50mL biocontrol composite bacterial agent every 5 days, and investigate the disease index, plant height, and plant fresh weight when the blank control is fully infected, and calculate the control effect.
[0040] 1 billion live spores / g Bacillus subtilis wettable powder (Baoding Kelvfeng Biochemical Technology Co., Ltd.) and 300 million CFU / g Trichoderma harzian wettable powder (Byward Co., Ltd., USA) were used as control agents. See Table 4 for the treatment and results of the pot test of Verticillium wilt in cotton.
[0041] Pot...
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