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

A detection method and resistance-related technology, which is applied in the field of CAPS markers to detect gene mutations related to abamectin resistance of spider mites, can solve problems such as short life history, drug resistance, and overlapping generations, and achieve sensitivity and accuracy High and specific effect

Inactive Publication Date: 2018-03-27
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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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 mite

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

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Embodiment Construction

[0024] 1. Materials and methods

[0025] 1. Source of tested insects

[0026]The Tetranychus urticae population that the present invention adopts comprises: indoor sensitive population (Lab-SS) and Beijing Miyun (MY-BJ), Beijing Shunyi (SY-BJ), Beijing Changping population (CP-BJ) and Zhejiang Ningbo population (NB -ZJ). Indoor Sensitive Population of Tetranychus urticae (Lab-SS) comes from the School of Plant Protection, Nanjing Agricultural University. Since 2009, the sponge leaf disk method has been used in an indoor artificial climate box, and insect-free bean seedling leaves of the "Bifeng" variety are used as hosts. Subculture, no contact with any pesticides during the period. The feeding conditions were 26±1°C, photoperiod L: D=16 h: 8 h. The four field populations were collected from April to July 2017 during the growth period of field horticultural crops in various places, and were reared indoors for one generation before being used for later use.

[0027] , test ...

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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...

Claims

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Application Information

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IPC IPC(8): C12Q1/6858
CPCC12Q1/6858C12Q2531/113C12Q2521/301
Inventor 王少丽徐丹丹何艳艳张友军吴青君谢文
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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