Molecular marker for resistance of panonychus citri to abamectin as well as application and detection method of molecular marker

A kind of citrus panclaw mite, molecular marker technology, applied in the field of molecular biology, to achieve the effect of rapid and effective mutation frequency, avoiding complexity and danger, and simple operation

Active Publication Date: 2021-01-05
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it is generally believed that the resistance of spider mites to abamectin is caused by the mutation of

Method used

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  • Molecular marker for resistance of panonychus citri to abamectin as well as application and detection method of molecular marker
  • Molecular marker for resistance of panonychus citri to abamectin as well as application and detection method of molecular marker
  • Molecular marker for resistance of panonychus citri to abamectin as well as application and detection method of molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Panonychus citrus is sensitive to avermectin detection

[0034] Sensitive strains: Indoor sensitive strains were collected from the nursery of the Citrus Research Institute of Southwest University in 2016 and raised in the laboratory for a long time for multiple generations without contact with any chemicals. 50 The value remained low at 0.010mg / L.

[0035] Field populations: The field populations of Panonychus citrus were selected from seven major citrus producing areas in southern my country: Sichuan (Daying, Anyue), Chongqing Wanzhou, Hubei Zigui, Guangxi (Nanning, Guilin) ​​and Yunnan Yuxi. The geographic location and host information are shown in Table 1 below. At least 10 trees were randomly selected in each park, more than 5,000 were collected for biological determination, and more than 500 female adult mites were collected at the same time and stored in absolute ethanol for subsequent DNA extraction. Bioassays were carried out on the sensitive strains...

Embodiment 2

[0041] Example 2 Field rapid detection of the resistance of Panonychus citrus to abamectin

[0042] Obtained in embodiment 1 from field Panonychus citrus in different areas collected from the field. Based on the position of the resistance mutation site of similar species, we tested the resistance markers of Panonychus citrus:

[0043] 1) Preparation of PCR amplification template: put the collected female adults of Panonychus citrus (over 500 heads) into a 1.5ml nuclease-free centrifuge tube as a sample, centrifuge at 12000rpm for 1min to the bottom of the tube, add 50μL STE Buffer (DNA extraction solution: 100mM NaCl, 10mM Tris-HCl, 1mM EDTA) and 2μL Proteinase K (50mg / ml), fully grind the mite body with a grinding rod, incubate in a water bath at 37°C for 30min, shake for 30s, then in a water bath at 95°C for 5min, 12000rpm After centrifugation for 3 minutes, the obtained supernatant was used as a template for PCR amplification.

[0044]2) PCR amplification: PCR reaction sy...

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Abstract

The invention discloses a molecular marker for resistance of panonychus citri to abamectin and application of the molecular marker in classification of populations sensitive to the abamectin and resistant to the abamectin of the panonychus citri. The molecular marker is PcGluCl1-G314E mutation or PcGluCl3-G310R mutation. The invention also discloses a specific primer pair for amplifying the molecular marker and a detection method of the molecular marker. The method comprises the following steps of extracting total DNA of panonychus citri to be detected, carrying out PCR amplification by usingthe specific primer pair by taking the total DNA as a template to obtain an amplification product GluCl1 or GluCl3 gene segment, analyzing whether the PCR amplification product contains the molecularmarker or not, and judging whether the panonychus citri to be detected is a sensitive population or a resistant population to abamectin. The molecular marker for the resistance of the panonychus citrito the abamectin is determined for the first time, and a rapid molecular detection means is provided for the field resistance of the panonychus citri to the abamectin.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and relates to a molecular marker, in particular to a molecular marker for the resistance of Panonychus citrus mite to abamectin and its application and detection method. Background technique [0002] Citrus Panonychus citri (McGregor), as a worldwide pest, occurs in large numbers every year, seriously affects the quality of citrus, and even threatens the healthy development of citrus industry. The control of Panonychus citrus still relies on chemical acaricides. Due to the characteristics of Panonychus citrus and the frequent application of pesticides in the field, it has developed resistance to a variety of acaricides and the resistance has developed rapidly. [0003] Abamectin is a natural product extracted from the fermented mycelium of Streptomyces avermitilis MA-4680 (NRRL8165), which has insecticidal, acaricidal and nematicidal activities, and is widely used in the agricultural f...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2600/124C12Q2531/113C12Q2565/125
Inventor 王进军潘登豆威袁国瑞刘巽燕
Owner SOUTHWEST UNIVERSITY
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