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Small brown rice planthopper drug-resistant carboxylesterase gene LSCE12, gene segment for lowering drug resistance of small brown rice planthoppers and application thereof

A technology of LSCE12 and carboxylesterase, which is applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of ineffective killing of pests, decrease of binding force between pesticides and targets, etc.

Active Publication Date: 2016-12-07
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 binding force between the insecticide and the target, which cannot effectively kill the pest, thus making the pest resistant to the insecticide

Method used

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  • Small brown rice planthopper drug-resistant carboxylesterase gene LSCE12, gene segment for lowering drug resistance of small brown rice planthoppers and application thereof
  • Small brown rice planthopper drug-resistant carboxylesterase gene LSCE12, gene segment for lowering drug resistance of small brown rice planthoppers and application thereof
  • Small brown rice planthopper drug-resistant carboxylesterase gene LSCE12, gene segment for lowering drug resistance of small brown rice planthoppers and application thereof

Examples

Experimental program
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Effect test

specific Embodiment

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

[0084] The deltamethrin-resistant strain used in the present invention was collected from Jianhu County, Jiangsu in 2009, and obtained by continuous screening of deltamethrin for 30 generations; the sensitive strain was collected from Jianhu County, Jiangsu in 2009, and was obtained without contacting any insecticides. Obtained by continuous breeding in the lower chamber.

[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 LSCE12 gene

[0088] The present invention obtains transcriptome data by constructing a transcriptome library of SBPH and performing high-throughput sequencing, and obtains LSCE12 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 Nanodrop1000 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

[0117] Example 2. Semi-quantitative analysis of LSCE12 gene in deltamethrin-resistant and sensitive strains of SBPH

[0118] The deltamethrin-resistant and sensitive materials of SBPH in the adult stage were selected, and the expression level of LSCE12 gene in these materials was analyzed by semi-quantitative RT-PCR, so that the relationship between the expression level of LSCE12 gene and deltamethrin resistance could be quickly obtained. relation.

[0119] (1) Extraction of total RNA from SBPH.

[0120] Using adult stage 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 gray fly The total RNA of lice 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 protein quantifier to confirm the quality and integri...

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Abstract

The invention relates to a small brown rice planthopper drug-resistant carboxylesterase gene LSCE12, a gene segment for lowering drug resistance of small brown rice planthoppers and application thereof. The sequence of the small brown rice planthopper drug-resistant carboxylesterase gene LSCE12 is disclosed as SEQ ID NO.1. The sequence of the gene segment for lowering drug resistance of small brown rice planthoppers is disclosed as SEQ ID NO.3. The gene segment is used for lowering drug resistance of small brown rice planthoppers. The gene segment can be used for detecting drug resistance of small brown rice planthoppers and implementing drug resistance treatment of small brown rice planthoppers.

Description

technical field [0001] The invention relates to a drug-resistant carboxylesterase gene LSCE12 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-strea...

Claims

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Application Information

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IPC IPC(8): C12N9/18C12N15/55C12N15/113C12N15/10C12N15/11
Inventor 徐鹿顾中言许小龙徐德进徐广春
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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