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Gene fragment of chitin synthetase 1 (SfCHS1) of Sogatella furcifera and application of dsRNA of gene fragment in pest control

A chitin synthase, white-backed planthopper technology, applied in DNA/RNA fragments, applications, genetic engineering, etc., can solve problems such as no related reports, no reports, and reduced survival rate.

Inactive Publication Date: 2018-11-16
GUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

By disrupting or blocking the formation of chitin synthase in insects, the phenomenon of molting disorder, reduced survival rate, and reduced fecundity in insects occurs.
Since chitin does not exist in humans and other higher animals, the key enzyme in the chitin synthesis process is relatively safe as a target for pest control. However, there is no relevant report on chitin synthase 1 in white-backed planthopper. , and there is no report on the dsRNA targeting the chitin synthase 1 gene of white-backed planthopper

Method used

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  • Gene fragment of chitin synthetase 1 (SfCHS1) of Sogatella furcifera and application of dsRNA of gene fragment in pest control
  • Gene fragment of chitin synthetase 1 (SfCHS1) of Sogatella furcifera and application of dsRNA of gene fragment in pest control

Examples

Experimental program
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Embodiment

[0014] Example: Chitin synthase 1 (SfCHS1) gene fragment and dsRNA of white-backed planthopper

[0015] 1. Obtaining the gene fragment of chitin synthase 1 (SfCHS1) from white-backed planthopper

[0016] 1) Preliminary analysis of the transcriptome database of the white-backed planthopper was carried out by using bioinformatics technology, and the partial nucleotide sequence of the SfCHS1 gene of the white-backed planthopper was determined through programs such as NCBI database Blast. Specific upstream primer 1 SEQ ID NO: 1 and downstream primer 1 SEQ ID NO: 2 were designed using Primer Premier 6.0 software, and all primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd.

[0017] 2) Select 15 5th-instar nymphs of white-backed planthopper, place them in a 1.5mL RNase-free centrifuge tube, freeze them in liquid nitrogen, grind them into fine powder quickly, and use the TRIzol method to extract total RNA. The specific operation steps refer to TRIzol kit instructio...

experiment example

[0028] Experimental example: dsRNA lethal experiment of chitin synthase 1 (SfCHS1) gene fragment of white-backed planthopper

[0029] 1. dsRNA injection of chitin synthase 1 (SfCHS1) gene fragment in white-backed planthopper

[0030] Nymphs with uniform size and health status on the first day of the 5th instar were selected for injection of the above-mentioned synthetic dsRNA. First prepare a 1% agarose plate, place the white-backed planthopper in CO 2 The treated lower abdomen was fixed upward on an agarose plate, and injected using a microinjector Nanoliter 2010 Injector (World Precision Instruments). The injection should be performed slightly, and the position where the prothorax and mid-thorax meet is used as the injection point. The amount of dsRNA injected was 100ng / head, and dsGFP was injected as the control group, 50 heads were injected in each group, and 3 biological replicates were set. After the injection, the test insects were put into test tubes containing fresh...

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Abstract

The invention discloses a gene fragment of chitin synthetase 1 (SfCHS1) of Sogatella furcifera and application of dsRNA of the gene fragment in pest control. A partial sequence of a chitin synthetase1 (SfCHS1) gene is obtained through analyzing a transcriptome database of Sogatella furcifera, cloning and sequencing are carried out on the basis of the partial sequence so as to obtain a nucleotidesequence as shown in SEQ ID NO: 3 in a sequence listing, dsRNA of the gene is synthesized on the basis of the sequence, i.e., SEQ ID NO: 3, thus, a candidate gene is provided for a Sogatella furciferaRNA interference mediated pest control strategy in the future, and meanwhile, a theoretical basis is provided for the research and development of novel insecticides. The invention belongs to the technical field of genetic engineering of insects.

Description

technical field [0001] The invention relates to the technical field of insect genetic engineering, in particular to the application of a white-backed planthopper chitin synthase 1 (SfCHS1) gene fragment and its dsRNA in pest control. Background technique [0002] The white-backed planthopper Sogatalla furcifera (Horváth) belongs to the Hemiptera planthopper family Delphacidae and is an important pest in rice production in the Asia-Pacific region. It mainly directly sucks the juice of rice plants through adults and nymphs, causing slow growth of rice, delayed tillering, increased shrunken grains, and can cause rice plants to die when seriously damaged; in addition, white-backed planthoppers are also southern rice black-streaked dwarf virus ( Southern rice black-streaked dwarfvirus, SRBSDV) is the main medium of transmission, and the harm it causes is even more devastating. At present, the prevention and control of white-backed planthopper still mainly depends on chemical ins...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N15/113A01P7/04
CPCC12N9/1051C12N15/1137C12N2310/14C12Y204/01016
Inventor 金道超王召杨洪周操龙贵云
Owner GUIZHOU UNIV
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