A Molecular Detection Method for Resistance of Plutella xylostella to Abamectin Targets

A technology for detection of abamectin targets and molecules, applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., can solve the problems of long cycle, low sensitivity, high material requirements, etc., and achieve high sensitivity , less material requirements, accurate detection effect

Inactive Publication Date: 2019-04-19
NANJING AGRICULTURAL UNIVERSITY
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve problems such as low sensitivity, long period, and high material requirements of the detection method of diamondback moth in the prior art for avermectin resistance. Rapid and Accurate Molecular Detection of Abamectin Resistance Target Mutations in Plutella xylostella

Method used

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  • A Molecular Detection Method for Resistance of Plutella xylostella to Abamectin Targets
  • A Molecular Detection Method for Resistance of Plutella xylostella to Abamectin Targets
  • A Molecular Detection Method for Resistance of Plutella xylostella to Abamectin Targets

Examples

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Embodiment 1

[0028] In this example, three groups of Plutella xylostella were selected for bioassay. Among them, the ROTH strain is a control strain that has been bred in the laboratory for many years and is sensitive to pesticides. In this example, the lethal concentration of the material to abamectin is taken as the baseline, and other The resistance level of the strain to abamectin; the ROTH-Abm strain is an established near-isogenic line material resistant to abamectin, and its resistance level to abamectin is 10,100 times; the Abm-Unsel strain is ROTH- Abm test insects were kept for more than one year without exposure to any chemicals, and its resistance was significantly reduced to 1695 times. The relevant biometric data are as follows:

[0029]

[0030] According to the preferred analysis and detection technology of the present invention, the specific implementation steps include:

[0031] 1. Randomly select 17, 19 and 26 fourth-instar larvae from ROTH, ROTH-Abm and Abm-Unsel strains, a...

Embodiment 2

[0046] This example illustrates the method for detecting the frequency of the 309 mutation gene of the PxGluCl gene carried by the Plutella xylostella Abm-Unsel strain of the present invention under different treatment concentrations.

[0047] 1. Extraction of the genomic DNA of single-headed diamondback moth larvae: Randomly pick the 3rd instar larvae of the diamondback moth of the Abm-Unsel strain, and treat a batch of test worms with 80ppm, 20ppm and 0ppm abamectin, respectively. The results are normal after 48h. Surviving larvae use AXYprep TM Multisource Genomic DNA Miniprep Kit for extraction of whole genome DNA.

[0048] 2. The surviving diamondback moth gene DNA templates (3 groups) after treatment with various concentrations of abamectin were used for PCR amplification of the target fragments of the glutamate chloride channel gene.

[0049] (1) Design specific primers for the glutamate chloride channel gene of Plutella xylostella, the upstream primer PxGluCl-Seq-F sequence...

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Abstract

The present invention discloses a molecular detection method of resistance of diamond back moth against an avermectin target. According to the method, a specific primer designed according a diamond back moth glutamate chloride ion channel gene sequence is used, extracted diamond back moth single-head genomic DNA is used as a template for PCR amplification of glutamate chloride ion channel gene fragments, diamond back moth individual genotypes (sensitive homozygote SS, resistant heterozygote RS and resistant homozygote RR) can be distinguished by analysis of a target fragment purified product direct sequencing chromatogram, and population sample resistance allele mutation frequency can be detected. A detection method of diamond back moth glutamate chloride ion channel gene A309V mutation and a special primer sequence are disclosed, and the method has the advantages of being simple, rapid and high in accuracy, can be used to monitor the occurrence and development trend of allele frequency of the resistance of a diamond back moth field population against avermectin and drug resistance, and provides an important basis for adjustment of diamond back moth chemical control strategies and formulation of measures to delay the development of resistance.

Description

Technical field [0001] The invention relates to a molecular detection method for the resistance of the diamondback moth to abamectin targets, belonging to the field of biotechnology, and is dedicated to the diamondback moth to the broad-spectrum insecticide and acaricide avermectin (composed of Avermectin B1a and Avermectin B1b) targets Highly sensitive, fast molecular detection of resistance. technical background [0002] According to statistics, in recent years, my country’s vegetable planting area has approached 300 million mu per year, and it has become the world’s largest cruciferous vegetable planting and producer. The reduction in vegetable production and the cost of treatment due to the damage of the diamondback moth are extremely high. Plutella xylostella belongs to the family Lepidoptera Plutella. It is an important cruciferous vegetable pest worldwide, causing economic losses of 4 to 5 billion US dollars each year. There are more than 40 kinds of host plants for this ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
Inventor 王兴亮吴益东杨亦桦武淑文
Owner NANJING AGRICULTURAL UNIVERSITY
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