CAPS marking method for detecting avermectin resistance related gene mutation of spider mites

Abstract

The invention discloses a CAPS marking method for detecting avermectin resistance related gene mutation of spider mites. Spider mite population GluCl3 gene to be detected which contains a gene fragment with G326E mutation is amplified, and enzyme sites are analyzed, wherein a Hinf I enzyme site exists in the amplified fragments of the sensitive population, and the site does not exist in the resistance homozygosis population, amplified fragments containing the Hinf I enzyme site exist in the resistance heterozygosis population, and amplified fragments without the site exist in the resistance heterozygosis population. The method can be used for replacing traditional biological assay methods, and has the advantages of little manpower and few sample amounts, large detection quantity, high sensitivity, good specificity, rapid and simplicity, and accuracy and reliability; the method is suitable for rapid diagnosis of avermectin resistance of Tetranychus urticae field population, and is usedfor carrying out real time monitoring and early warning forecast of avermectin resistance of field spider mites at the same time; the method has important meaning for avermectin resistance monitoringof field spider mites, and application prospect is wide.

Description

technical field [0001] The invention belongs to the technical field of biotechnology, and in particular relates to a CAPS marking method for detecting gene mutations related to the resistance of spider mites to abamectin. Background technique [0002] Two-spotted spider mite Tetranychus urticae Koch is a major agricultural pest worldwide on fruit trees, vegetables and other crops. Tetranychus urticae sucks the juice of plants by sucking adult mites and nymphs. The leaves turn yellow in the early stage and affect photosynthesis. In severe cases, the leaves die, the leaves fall or even the whole plant dies, which seriously affects the healthy growth of crops and product quality. [0003] Due to its small size, short life cycle, and serious overlapping of generations, the frequent use of chemical agents in the field has led to the emergence of drug resistance of Tetranychus urticae. It has been reported that field populations of Tetranychus urticae have developed varying deg...

AI Technical Summary

Problems solved by technology

[0003] Due to its small size, short life cycle, and serious overlapping of generations, the frequent use of chemical agents in the field has led to the emergence of drug resistance of the two-spotted spider mite

Conventional monitoring methods for insecticide resistance of pest mites are based on bioassay methods, such as slide immersion method recommended by FAO, which is widely used, but this method has many disadvantages: (1) The operation process is very delicate and requires extreme care to ensure that the test Spider mites are not harmed; (2) It takes a long time for the operator to master the technology; (3) The bioassay process requires 48 hours after the chemical treatment to observe the results and judge the mortality of pest mites

The PCR-based point mutation detection method needs to go through the following steps: gene amplification, fragment sequencing and sequence comparison to determine whether the resistance-related gene has mutated, but it is obvious that such a process is time-consuming and laborious. It also consumes more resources

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