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Molecular marker for resistance of Panonychus citri (McGregor) to pyridaben as well as application and detection method of molecular marker

A citrus panonychia, molecular marker technology, applied in the field of molecular biology, to avoid harm, eliminate time-consuming and labor-intensive, and avoid complex and dangerous effects

Pending Publication Date: 2021-02-05
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Complex I, also known as NADH dehydrogenase (NADHdehydrogenase), is located in the inner membrane of mitochondria and is the largest complex among the five complexes of the electron transport chain. Among them, PSST is a subunit of complex I. There is no molecule yet Markers can rapidly detect the resistance of Panonychus citrus to pyridaben

Method used

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  • Molecular marker for resistance of Panonychus citri (McGregor) to pyridaben as well as application and detection method of molecular marker
  • Molecular marker for resistance of Panonychus citri (McGregor) to pyridaben as well as application and detection method of molecular marker
  • Molecular marker for resistance of Panonychus citri (McGregor) to pyridaben as well as application and detection method of molecular marker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Sensitive strains: Indoor sensitive strains were collected from the nursery of the Citrus Research Institute of Southwest University in 2016, and were raised in the laboratory for multiple generations without contact with any chemicals during the period. The LC of pyridaben 50 The value remained low at 0.131mg / L.

[0033] Field populations: The field populations of Panonychus citrus were selected from 7 main citrus producing areas in southern my country: Sichuan (Daying, Anyue), Chongqing Wanzhou, Hubei Zigui, Guangxi (Nanning, Guilin) ​​and Yunnan Yuxi. At least 10 trees were randomly selected in each park, and more than 5,000 were collected for biological determination, and 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 and the aforementioned 7 field populations. The results are shown in Table 1. Compared with the indoor sensitive strains, the Panonychu...

Embodiment 2

[0036] Example 2 Field rapid detection of the resistance of Panonychus citrus to pyridaben

[0037] Using Example 1 to obtain Panonychus citrus in different areas collected from the field, according to the position of the similar species resistance mutation site, the resistance marker detection of Panonychus citrus is carried out:

[0038] 1) Preparation of PCR amplification template: put the collected adult females of Panonychus citrus into a 1.5ml nuclease-free centrifuge tube as a sample, centrifuge at 12000rpm for 1min to the bottom of the tube, add 20μL of STE Buffer (DNA extraction Solution: 100mM NaCl, 10mM Tris-HCl, 1mM EDTA) and 0.5μL 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 10s, then in a water bath at 95°C for 5min, centrifuge at 12000rpm for 3min, The obtained supernatant was used as a template for PCR amplification.

[0039] 2) PCR amplification: PCR reaction system: 12.5 μL 2×Pri...

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Abstract

The invention discloses a molecular marker for resistance of Panonychus citri (McGregor) to pyridaben and application of the molecular marker in classification of sensitive populations and resistant populations of the Panonychus citri (McGregor) to the pyridaben. The molecular marker is PcPSST_H107R mutation. The invention also discloses a specific primer pair for amplifying the molecular marker.The primer pair is PcPSST_F / R. The invention also discloses a detection method of the molecular marker for the resistance of the Panonychus citri (McGregor) to the pyridaben. The method comprises thefollowing steps of firstly extracting the total DNA of the Panonychus citri (McGregor) to be detected, carrying out PCR amplification by using the specific primer pair through the total DNA as a template to obtain an amplification product PSST gene segment, and analyzing whether the PCR amplification product contains the molecular marker or not, thereby judging whether the Panonychus citri (McGregor) to be detected is the sensitive population or the resistant population to the pyridaben. According to the invention, the molecular marker for the resistance of the Panonychus citri (McGregor) to the pyridaben is determined for the first time, and the method for rapidly detecting the molecular marker for the resistance of the Panonychus citri (McGregor) to the pyridaben is established.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a resistance gene molecular marker, in particular to a molecular marker for the resistance of Panonychus citrus mite to pyridaben and its application and detection method. Background technique [0002] The citrus mite Panonychus citri (McGregor) is an important worldwide occurrence and damage pest mite, which can eat and damage 112 kinds of host plants, but mainly citrus fruit trees. The mite is widely distributed in citrus producing areas such as Asia, Europe, Africa, and North and South America. It has multiple generations a year, and it occurs seriously during the citrus shoot period from April to June and September to November every year. At present, the control of Panonychus citrus is still dominated by chemical control. However, due to long-term large-scale and irrational drug use, coupled with the characteristics of Panonychus citrus, such as strong fecundity, short devel...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/53C12N15/11C12Q1/6888C12Q1/6858
CPCC12N9/0036C12Y106/99003C12Q1/6888C12Q1/6858C12Q2600/156C12Q2600/124C12Q2600/106C12Q2531/113C12Q2565/125
Inventor 王进军潘登豆威袁国瑞牛金志刘巽燕
Owner SOUTHWEST UNIVERSITY
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