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A method for improving the insecticidal activity of Bt by mixing with RNAi engineering bacteria
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A technology of insecticidal activity and engineering bacteria, which is applied in the field of agricultural biology, can solve the problems of difficult application of field pests, and achieve the effect of improving lethal efficiency and insecticidal activity
Active Publication Date: 2022-05-24
FUJIAN AGRI & FORESTRY UNIV
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Problems solved by technology
In early RNAi research, the effect of gene silencing was generally achieved through injection, mixed food feeding, etc., but the application of these methods in field pest control is relatively difficult
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Embodiment 1
[0021] 1. Pick the Bt 8010 stored in the laboratory and the 8010AKi engineering bacteria that have been constructed (ie 8010-AKi in Chen Jinzhi's master's thesis "Bacillus thuringiensis-mediated RNA interference of the arginine kinase gene of diamondback moth"), 30 After overnight incubation at 200 rpm at °C, diluted to 1 OD with sterile water, and then mixed at a volume ratio of 10:0, 9:1, 7:3, 5:5, 3:7, 1:9 and 0:10.
[0022] 2. Cut the Plutella xylostella artificial feed into squares of about 10 cm×10 cm×10 cm, soak them in the mixed bacterial solution for 2 h, and dry them.
[0023] 3. Take 30 third-year-old diamondback moths after starvation treatment for 4 hours, put them in the rearing box, and put them in the air-dried feed. Record the number of deaths of diamondback moths every 12 hours, and record the number of diamondback moth pupae every 24 hours. Weight, artificial feed was changed every 24 h. There were 3 biological replicates for each treatment.
[0024] 4. Co...
Embodiment 2
[0035] 1. Pick the Bt 8010 stored in the laboratory and the BMB171AKi engineering bacteria that have been constructed (ie BMB171-AKi in Chen Jinzhi's master's thesis "Bacillus thuringiensis-mediated RNA interference of the diamondback moth arginine kinase gene"), 30 After overnight incubation at 200 rpm at °C, dilute to 1 OD with sterile water, and then mix at volume ratios of 10:0, 9:1, 7:3, 5:5, 3:7, 1:9, and 0:10 .
[0036] 2. Cut the Plutella xylostella artificial feed into squares of about 10 cm×10 cm×10 cm, soak them in the mixed bacterial solution for 2 h, and dry them.
[0037] 3. Take 30 third-year-old diamondback moths after starvation treatment for 4 hours, put them in the rearing box, and put them in the air-dried feed. Record the number of deaths of diamondback moths every 12 hours, and record the number of diamondback moth pupae every 24 hours. Weight, artificial feed was changed every 24 h. There were 3 biological replicates for each treatment.
[0038] 4. Co...
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Abstract
The invention belongs to the field of agricultural biotechnology, and relates to a method for improving the insecticidal activity of Bt by mixing with RNAi engineering bacteria. The Bacillus thuringiensis engineering bacteria 8010AKi and BMB171AKi which can interfere with the arginine kinase gene of diamondback moth are selected, and the engineering bacteria are respectively After mixing with Bacillus thuringiensis 8010 in proportion, the diamondback moth was fed. The test shows that when 8010 and 8010AKi are fed in a ratio of 9:1, and BMB171AKi is fed in a ratio of 7:3 to Plutella xylostella, both can significantly increase the lethality of Bacillus thuringiensis 8010 to Plutella xylostella, and the mortality of Plutella xylostella respectively reached 68.9% and 73.1%, indicating that a certain proportion of mixing can indeed improve the insecticidal activity of Bt, which expands the direction for the application of Bt biopesticide preparations in the future, and also provides a new idea for the application of RNAi engineering bacteria.
Description
technical field [0001] The invention belongs to the technical field of agricultural biotechnology, and relates to a method for improving the insecticidal activity of Bt by mixing with RNAi engineering bacteria. In the present invention, Bacillus thuringiensis engineering bacteria 8010AKi and BMB171AKi expressing the arginine kinase gene dsRNA of Plutella xylostella are selected, and the engineering bacteria are respectively mixed with Bacillus thuringiensis 8010 in proportions of 10:0, 9:1, 7:3, 5:5, 3 :7, 1:9 and 0:10, mixed with feed and fed to diamondback moth. It is expected to obtain a method that can improve the insecticidal activity of Bacillus thuringiensis 8010 by mixing with RNAi engineered bacteria. Background technique [0002] Bacillus thuringiensis ( Bacillus thuringiensis , Bt for short) is a widely distributed gram-positive bacillus, which produces parasporal crystals with insecticidal activity at one or both ends of the spore during spore formation, and pa...
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