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Method for improving insecticidal activity of Bt by mixing with RNAi engineering microorganisms

A technology of insecticidal activity and engineering bacteria, applied in the field of agricultural biology, can solve problems such as difficulties in the application of field pests

Active Publication Date: 2020-10-20
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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

Method used

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  • Method for improving insecticidal activity of Bt by mixing with RNAi engineering microorganisms
  • Method for improving insecticidal activity of Bt by mixing with RNAi engineering microorganisms
  • Method for improving insecticidal activity of Bt by mixing with RNAi engineering microorganisms

Examples

Experimental program
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Effect test

Embodiment 1

[0021] 1. Pick the Bt 8010 preserved in the laboratory and the 8010AKi engineering bacteria that have been constructed (that is, the 8010-AKi in Chen Jinzhi's master's thesis "Bacillus thuringiensis-Mediated RNA Interference of Plutella xylostella Arginine Kinase Gene"), 30 After culturing overnight at 200 rpm at ℃, dilute to 1 OD with sterile water, then mix according to volume ratios of 10:0, 9:1, 7:3, 5:5, 3:7, 1:9 and 0:10 .

[0022] 2. Cut the artificial diet of Plutella xylostella into cubes with a size of about 10 cm×10 cm×10 cm, soak it in the mixed bacterial solution for 2 h, and dry it in the air.

[0023] 3. Take 30 third-instar diamondback moths and starve them for 4 hours, put them in a breeding box, put them in dry feed, record and count the number of dead diamondback moths every 12 hours, and record the number of dead diamondback moths every 24 hours. The artificial diet was changed every 24 h. Each treatment was biologically repeated 3 times.

[0024] 4. Col...

Embodiment 2

[0035] 1. Pick the Bt 8010 preserved in the laboratory and the BMB171AKi engineering bacteria that have been constructed (that is, BMB171-AKi in Chen Jinzhi's master's thesis "Bacillus thuringiensis-mediated RNA interference of diamondback moth arginine kinase gene"), 30 After culturing overnight at 200rpm at ℃, dilute to 1 OD with sterile water, and then mix according to volume ratios of 10:0, 9:1, 7:3, 5:5, 3:7, 1:9 and 0:10.

[0036] 2. Cut the artificial diet of Plutella xylostella into cubes with a size of about 10 cm×10 cm×10 cm, soak it in the mixed bacterial solution for 2 h, and dry it in the air.

[0037] 3. Take 30 third-instar diamondback moths and starve them for 4 hours, put them in a breeding box, put them in dry feed, record and count the number of dead diamondback moths every 12 hours, and record the number of dead diamondback moths every 24 hours. The artificial diet was changed every 24 h. Each treatment was biologically repeated 3 times.

[0038] 4. Colle...

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Abstract

The present invention belongs to the technical field of agricultural biology and relates to a method for improving insecticidal activity of Bt by mixing with RNAi engineering microorganisms. The engineering microorganisms 8010AKi and BMB171AKi of bacillus thuringiensis engineering microorganisms are selected to interfere with plutella xylostella arginine kinase gene, and the engineering microorganisms are respectively mixed with bacillus thuringiensis 8010 in proportion to feed plutella xylostella. Experiments show that when the 8010 and 8010Aki are mixed at a ratio of 9:1 and when the 8010 and BMB171Aki are mixed at a ratio of 7:3 to feed the plutella xylostella, the mixtures can both significantly increase lethality of the bacillus thuringiensis 8010 against the plutella xylostella, andthe mortality of the plutella xylostella respectively reaches 68.9% and 73.1%, indicating that a certain proportion of the mixture can indeed improve the insecticidal activity of the Bt, which expandsdirection for future application of Bt biopesticide preparations and also provides new ideas for application of the RNAi engineering microorganisms.

Description

technical field [0001] 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 present invention selects Bacillus thuringiensis engineering bacteria 8010AKi and BMB171AKi expressing Plutella xylostella arginine kinase gene dsRNA, and the engineering bacteria and Bacillus thuringiensis 8010 are proportioned 10:0, 9:1, 7:3, 5:5, 3 respectively. :7, 1:9 and 0:10 mixed with the feed and then fed to the diamondback moth. It is expected to obtain a method that can improve the insecticidal activity of Bacillus thuringiensis 8010 by mixing with RNAi engineering bacteria. Background technique [0002] Bacillus thuringiensis ( Bacillus thuringiensis , referred to as Bt) is a widely distributed Gram-positive bacillus, which produces parasporal crystals with insecticidal activity at one or both ends of the bacterium during spore formation, and the parasporal cr...

Claims

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Application Information

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IPC IPC(8): C12N15/75C12N15/54A01N63/23A01P7/04
CPCC12N15/75C12N9/1223A01N63/23C12Y207/03003
Inventor 杨广陈金芝蒋颖霞李妙雯张彤傅淑
Owner FUJIAN AGRI & FORESTRY UNIV