Drug-resistant gene CYP6AY3v2 of small brown planthopper, gene segment for reducing drug resistance of small brown planthopper and application thereof

A technology of gene fragments and SBPH, applied in the direction of DNA/RNA fragments, applications, genetic engineering, etc., can solve the problems of ineffective killing of pests, decreased binding force between pesticides and targets, etc.

Active Publication Date: 2016-09-28
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Abstract
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  • Claims
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Problems solved by technology

Target resistance is due to changes in the conformation of the target of the insecticide in the body, resulting in a decrease in the bindi

Method used

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  • Drug-resistant gene CYP6AY3v2 of small brown planthopper, gene segment for reducing drug resistance of small brown planthopper and application thereof
  • Drug-resistant gene CYP6AY3v2 of small brown planthopper, gene segment for reducing drug resistance of small brown planthopper and application thereof
  • Drug-resistant gene CYP6AY3v2 of small brown planthopper, gene segment for reducing drug resistance of small brown planthopper and application thereof

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specific Embodiment

[0083] The present invention is further described below by embodiment, but the present invention is not limited to these examples.

[0084] The chlorpyrifos and deltamethrin-resistant strains used in the present invention were collected from Jianhu County, Jiangsu Province in 2009, and obtained by continuous screening of chlorpyrifos and deltamethrin for 25 generations and 30 generations respectively; the sensitive strains were collected from Jianhu County, Jiangsu Province in 2009. Obtained by continuous breeding indoors without exposure to any pesticides.

[0085] The reagents involved in the present invention are all commercially available.

[0086] The experimental methods of the following examples are conventional methods unless otherwise specified.

Embodiment 1

[0087] Example 1. Cloning and sequence analysis of CYP6AY3v2 gene

[0088] The present invention obtains transcriptome data by constructing a transcriptome library of SBPH and performing high-throughput sequencing, and obtains CYP6AY3v2 gene fragments on this basis.

[0089] (1) Extraction of total RNA from SBPH.

[0090] Using sensitive strains of SBPH as research materials, select SBPH eggs, 1-5 instar nymphs, and male and female adults for mixed extraction using TRIzol Total RNA Extraction Kit (Shanghai Yingjun Biotechnology Co., Ltd.), and the operation method refers to the reagents box instructions. The extracted total RNA of SBPH was electrophoresed on agarose gel with a concentration of 1.0%, and the RNA concentration and absorbance value were detected with a Nanodrop100 nucleic acid protein quantifier to confirm the quality and integrity of the RNA, and then placed in an ultra-low temperature refrigerator at Freeze at -80°C.

[0091] (2) Preparation of cDNA template...

Embodiment 2

[0120] Example 2. Semi-quantitative analysis of CYP6AY3v2 gene in chlorpyrifos and deltamethrin resistant and sensitive strains of SBPH

[0121] Select chlorpyrifos and deltamethrin-resistant and sensitive materials of SBPH in the adult stage, and use semi-quantitative RT-PCR to analyze the expression level of CYP6AY3v2 gene in these materials, so as to quickly obtain the expression level of CYP6AY3v2 gene and the relationship between chlorpyrifos and deltamethrin Ester resistance relationship.

[0122] (1) Extraction of total RNA from SBPH.

[0123] Using adult chlorpyrifos and deltamethrin-resistant and sensitive SBPH as materials, total RNA was extracted using the SV Total RNA extraction kit (Promega (Beijing) Biotechnology Co., Ltd.) according to the operating procedures, and the extracted The total RNA of SBPH was electrophoresed on agarose gel with a concentration of 1.0%, and the concentration and absorbance of the RNA were detected with a Nanodrop100 nucleic acid prot...

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Abstract

The invention relates to a drug-resistant gene CYP6AY3v2 of a small brown planthopper, a gene segment for reducing the drug resistance of the small brown planthopper and application thereof. A sequence of the drug-resistant gene CYP6AY3v2 of the small brown planthopper is shown as SEQ ID NO.1. A sequence of the gene segment for reducing the drug resistance of the small brown planthopper is shown as SEQ ID NO.3. The gene segment is used for reducing the drug resistance of the small brown planthopper. The drug-resistant gene CYP6AY3v2 and the gene segment can be used for detecting the drug resistance of the small brown planthopper and achieving the drug resistance management of the small brown planthopper.

Description

technical field [0001] The invention relates to a drug resistance gene CYP6AY3v2 of the planthopper, a gene fragment for reducing the drug resistance of the planthopper and an application thereof, and belongs to the field of biotechnology. Background technique [0002] As far as the applicant knows, the gray planthopper Laodelphax striatellus (Fallén) belongs to Hemiptera, planthopper family, and is mainly distributed in the Asian rice region from the Philippines to Siberia, and the temperate and subtropical regions of Europe. In my country, SBPH are distributed in all provinces and cities, especially in the Yangtze River Basin and North China rice fields. SBPH is a polyphagous insect that can feed on a variety of gramineous plants such as rice, wheat, and corn. Apart from sucking the juice of the host plant with its own piercing-sucking mouthparts, the main damage form of SBPH is the transmission of plant virus diseases, such as stripe leaf blight, black-streaked dwarf dis...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N9/02C12N15/113C12N15/10C12N15/11A01P7/04
CPCC12N9/0071C12N15/11C12N15/1137C12N2310/14C12N2330/50
Inventor 徐鹿韩召军顾中言徐德进
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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