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Application of the brown planthopper nlinr gene as a target in the preparation of drugs for the control of brown planthopper

A brown planthopper and drug technology, applied in application, genetic engineering, DNA/RNA fragments, etc., can solve the problems of insecticide resistance, residue and pest re-emergence, and achieve ovarian development retardation, inhibition of reproduction, and reduction of egg production. Effect

Active Publication Date: 2021-06-29
HANGZHOU NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the long-term irrationality of such pesticides has brought about disadvantages such as the "3R" problem (resistance, residue, and pest re-emergence). Based on the interests of high-quality and high-yield rice, green prevention and control will become a major issue in modern agricultural production. the necessary trend of

Method used

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  • Application of the brown planthopper nlinr gene as a target in the preparation of drugs for the control of brown planthopper
  • Application of the brown planthopper nlinr gene as a target in the preparation of drugs for the control of brown planthopper
  • Application of the brown planthopper nlinr gene as a target in the preparation of drugs for the control of brown planthopper

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Acquisition of the brown planthopper insulin receptor genes NlInR1 and NlInR2

[0054] 1. Extraction of RNA:

[0055] The RNA of brown planthopper reared indoors (feeding conditions: temperature 25±1°C, humidity 70%, light ratio 16L:8D) was extracted by Trizol method, and used as template RNA.

[0056] Single-strand cDNA synthesis: 1 μg template RNA was used to synthesize single-strand cDNA using AMV reverse transcription kit (Takara, Baosheng, Dalian) as a template for amplifying the middle fragment sequence.

[0057] 2. Amplification of the NlInR gene target fragment sequence

[0058] Primers were designed according to the N. lugens cDNA sequence, and the primer sequences were as follows:

[0059] NlInR1-F: 5'-CGCTCTGGTTGTGCTTGATA-3';

[0060] NlInR1-R: 5'-CGTTGTCTTTCTCCAACGGT-3';

[0061] NlInR2-F: 5'-AGCTGTGCAGGGAGAATGTT-3';

[0062] NlInR2-R: 5'-AGTCCCTTGGTACCTTTTCCG-3';

[0063] Use the above primers for PCR amplification. The system is as follows:...

Embodiment 2

[0065] Example 2 Application of NlInR in pest control

[0066] 1. Synthesis of dsRNA

[0067] Carry out the amplification of InR1, InR2 and GFP (Green Fluorescent Protein: Green Fluorescent Protein) target fragments, gel recovery, connection, transformation with the method of Example 1, and carry out PCR identification, then carry out sequencing, transform the sequenced correct transformant into containing Amp+ (100 μg / ml) LB liquid medium, 250 rpm, 37° C. shake culture overnight, and use the plasmid drawer kit to extract the plasmid. Then use T7 The Express RNAi System kit (Promega) synthesized dsInR1, dsInR2 and dsGFP, and mixed dsInR1 and dsInR2 at a ratio of 1:1 to form dsInRs. For specific operations, refer to the T7RiboMAX Express RNAi System kit manual. The primer sequences used are as follows:

[0068] DSNlInR1-F: 5'-CGCTCTGGTTGTGCTTGATA-3';

[0069] DSNlInR1-R: 5'-CGTTGTCTTTCTCCAACGGT-3';

[0070] DSNlInR2-F: 5'-AGCTGTGCAGGGAGAATGTT-3';

[0071] DSNlInR2-R: 5'-A...

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Abstract

The invention discloses the application of a brown planthopper NlInR gene as a target in the preparation of drugs for controlling brown planthopper, and provides a dsRNA that inhibits the expression of the NlInR gene. The NlInR gene is composed of the NlInR1 gene and the NlInR2 gene. The acid sequence is shown in SEQ ID NO.1, and the nucleotide sequence of the NlInR2 gene is shown in SEQ ID NO.2. The present invention discovers for the first time the role of the NlInR gene in the prevention and treatment of brown planthoppers. The dsRNA of the NlInR gene is obtained by RNAi technology, and after the NlInR gene is silenced by microinjection, it can effectively inhibit the individual development and ovarian development of the brown planthopper and reduce the amount of eggs laid, thereby Inhibit the reproduction of brown planthopper and realize the control of brown planthopper.

Description

technical field [0001] The invention relates to the technical field of prevention and control of agricultural diseases and insect pests, in particular to the application of the brown planthopper NlInR gene as a target in the preparation of medicines for controlling brown planthopper. Background technique [0002] As a big rice country in the world, our country is one of the countries where the occurrence of crop pests is the most serious. In order to control the loss and damage caused by pests, chemical pesticides and insecticides have become a very important choice due to their rapid and effective control characteristics, forming a situation where chemical control is more important than comprehensive control. However, the long-term irrationality of such pesticides has brought about disadvantages such as the "3R" problem (resistance, residue, and pest re-emergence). In the interests of high-quality and high-yield rice, green prevention and control will become an important pa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113A01N57/32A01P7/04
CPCA01N57/32C12N15/1138C12N2310/14
Inventor 罗雨嘉刘永康邓颖梅庞晓青王世贵唐斌
Owner HANGZHOU NORMAL UNIVERSITY
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