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Fast detection method of kdr (Knockdown Resistance) caused by mutation of tetranychus cinabarinus boisdu F1538I

A technology of Tetranychus cinnabarinus and a detection method, which is applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of inaccurate detection of genotype composition, complicated operation, and many biological test materials.

Inactive Publication Date: 2014-04-16
SOUTHWEST UNIVERSITY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Conventional detection methods have the disadvantages of requiring many biological test materials, complex operation, and inability to accurately detect the genotype composition of pest field kdr resistance.

Method used

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  • Fast detection method of kdr (Knockdown Resistance) caused by mutation of tetranychus cinabarinus boisdu F1538I
  • Fast detection method of kdr (Knockdown Resistance) caused by mutation of tetranychus cinabarinus boisdu F1538I

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Embodiment

[0033] Using the method of allelic PCR, design specific primers for kdr mutation genotype detection according to the cDNA sequence of Tetranychus cinnabarinus sodium ion channel: Primer1 and Primer2 are used to amplify non-mutant fragments of about 150bp, and Primer3 and Primer4 are used to amplify 600bp Left and right mutant fragments. Genomic DNA of Tetranychus cinnabarinus was extracted from a single head, and its kdr mutation genotype was detected by electrophoresis after PCR.

[0034] PCR amplification using primers

[0035] Primer name sequence S-Forward 5'-TTCAAGTGGCAACATTCAAAGG-3' S-Reverse 5'-AATAAACAGGTTAAGTGTGAA-3' R-Forward 5'-TTCATCATTTTTGGCTCTTTTA-3' R-Reverse 5'-TGGTCAAGGGACATCACA-3'

[0036] Instruments and reagents used

[0037] Instruments: a set of precision micropipettes, PCR instrument, electrophoresis tank, microwave oven, gel imaging analyzer

[0038] Consumables: PCR tube, 1.5mL centrifuge tube, pipette tip...

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Abstract

The invention relates to a fast detection method of kdr (Knockdown Resistance) caused by the mutation of tetranychus cinabarinus boisdu F1538I. According to the fast detection method disclosed by the invention, PCR (Polymerase Chain Reaction) detection is carried out aiming to the mutation of the kdr (Knockdown Resistance) of a tetranychus cinabarinus boisdu sodion channel gene, and the target resistance (kdr) of the tetranychus cinabarinus boisdu to pyrethroid insecticides and miticides can be fast monitored by virtue of primer design, extraction of a tetranychus cinabarinus boisdu genome DNA (Deoxyribonucleic Acid), PCR (Polymerase Chain Reaction) amplification and the like. With the adoption of the fast detection method, a great amount of biological testing pieces can be saved, the operation is simple, and a result is fast and definite; and the fast detection method can be used for replacing conventional biological determination method used for grasping the drug resistance occurrence and development trends of the tetranychus cinabarinus boisdu to the pyrethroid insecticides and miticides.

Description

technical field [0001] The invention relates to the technical field of plant protection, in particular to a rapid detection method for kdr resistance caused by the F1538I mutation of Tetranychus cinnabarinus. Background technique [0002] Tetranychus cinnabarinus, also known as cotton spider mite, cotton spider mite, red spider mite, is widely distributed in my country, and occurs in North China, East China, Central China, South China, Northeast China, Yunnan, Gansu and other regions. There are 146 species of plants in 43 families that can be harmful, mainly beans, eggplant, sorghum, pepper, watermelon, strawberry, corn, onion, amaranth, mulberry and weeds. The mite has a very strong reproductive ability. It lays a lot of eggs in a suitable growth environment and has a short generation cycle. It can quickly form a certain number of populations in a short period of time, and then gather on the back of the plant leaves and both sides of the veins. It pricks and sucks the plan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q2531/113
Inventor 何林徐志峰刘炎超钱坤
Owner SOUTHWEST UNIVERSITY
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