Application of NlInR gene of brown planthopper as target point in preparation of pesticide for preventing and treating brown planthopper

A technology for brown planthoppers and transgenic plants, applied in applications, genetic engineering, DNA/RNA fragments, etc., can solve problems such as insecticide resistance, residues, and pest re-emergence, and achieve smaller body size, lower egg production, and inhibition reproductive effect

Active Publication Date: 2019-07-30
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 o

Method used

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  • Application of NlInR gene of brown planthopper as target point in preparation of pesticide for preventing and treating brown planthopper
  • Application of NlInR gene of brown planthopper as target point in preparation of pesticide for preventing and treating brown planthopper
  • Application of NlInR gene of brown planthopper as target point in preparation of pesticide for preventing and treating brown planthopper

Examples

Experimental program
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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 application of a NlInR gene of brown planthopper as a target point in preparation of a pesticide for preventing and treating the brown planthopper, and provides dsRNA used forinhibiting expression of the NlInR gene, wherein the NlInR gene is composed of a NlInR1 gene and a NlInR2 gene; the nucleotide sequence of the NlInR1 gene is shown as SEQ ID NO.1; the nucleotide sequence of the NlInR2 gene is shown as SEQ ID NO.2. The effect of the NlInR gene in preventing and treating the brown planthopper is discovered for the first time, the dsRNA of the NlInR gene is obtainedthrough an RNAi technology, and after the NlInR gene is silenced through a micro-injection mode, the individual development and the ovarian development of the brown planthopper can be effectively inhibited, and the spawning amount is reduced, so that the propagation of the brown planthopper is inhibited, and the prevention and treatment of the brown planthopper are realized.

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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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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