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Sogatalla furcifera chitin synthase 1a (SfCHSla) gene alternative spliceosome and application of its dsRNA 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 relevant reports, no dsRNA, and reduced survival rate.

Inactive Publication Date: 2018-11-06
KAILI 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 enzymes in the process of chitin synthesis are relatively safe targets for pest control. However, there are no related reports on chitin synthase 1a in white-backed planthopper. , and there is no report on the dsRNA targeting the chitin synthase 1a gene of white-backed planthopper

Method used

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  • Sogatalla furcifera chitin synthase 1a (SfCHSla) gene alternative spliceosome and application of its dsRNA in pest control
  • Sogatalla furcifera chitin synthase 1a (SfCHSla) gene alternative spliceosome and application of its dsRNA in pest control

Examples

Experimental program
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Embodiment

[0014] Example: Variable splicer of chitin synthase 1a (SfCHS1a) gene of white-backed planthopper and its dsRNA

[0015] 1. Acquisition of variable splices of chitin synthase 1a (SfCHS1a) gene in white-backed planthopper

[0016] 1) Based on the nucleotide sequence of the white-backed planthopper chitin synthase 1a with the accession number KY350143 in the GenBank database, use Primer Premier 6.0 software to design specific upstream primer 1 SEQ ID NO: 1 and downstream primer 1 SEQ ID NO : 2. 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 instructions. Then, the first-strand cDNA was synthesized according to the instructions of the AMV First Strand cDNA Synthesis Kit ...

experiment example

[0027] Experimental example: dsRNA lethal experiment of the variable splice of chitin synthase 1a (SfCHS1a) gene of white-backed planthopper

[0028]1. dsRNA injection of the variable splice of chitin synthase 1a (SfCHS1a) gene in white-backed planthopper

[0029] 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 i...

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Abstract

The invention discloses a sogatalla furcifera chitin synthase 1a (SfCHSla) gene alternative spliceosome and application of its dsRNA in pest control and belongs to the technical field of insect genetic engineering. The SfCHS1a gene alternative splicesome is obtained on the basis of the nucleotide sequence of SfCHS1a of which the accession number in a GenBank database is KY350143, wherein, the nucleotide sequence is as shown as SEQ ID NO:3. Further, the dsRNA of the gene is synthesized on the basis of the sequence SEQ ID NO:3. Accordingly, The candidate gene is provided for sogatalla furciferaRNA mediated pest control strategies, and meanwhile, a theoretical basis is provided for research and development of novel insecticides.

Description

technical field [0001] The invention relates to the technical field of insect genetic engineering, in particular to the application of a variable splicer of chitin synthase 1a (SfCHS1a) gene of white-backed planthopper 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 dwarf virus, 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 depend...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12N15/10C12N15/113A01N57/16A01P7/04
CPCA01N57/16C12N9/1051C12N2310/14C12Y204/01016
Inventor 王召杨洪朱环金道超周操龙贵云
Owner KAILI UNIV
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