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
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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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