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Sogatella furcifera phosphorus acetylglucosamine mutase gene fragment and application thereof

A technology of phosphorus acetamido and gene fragments of white-backed planthopper, applied in the fields of application, genetic engineering, plant gene improvement, etc. health threats

Inactive Publication Date: 2019-01-22
KAILI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a gene fragment of phosphoacetylglucosamine mutase of white-backed planthopper and its application, to solve the problem that the current prevention and control of white-backed planthopper still mainly depends on chemical insecticides, and the A large number of unreasonable use poses a serious threat to the ecological environment and human health, and also makes white-backed planthoppers produce varying degrees of resistance to various insecticides

Method used

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  • Sogatella furcifera phosphorus acetylglucosamine mutase gene fragment and application thereof
  • Sogatella furcifera phosphorus acetylglucosamine mutase gene fragment and application thereof

Examples

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Embodiment 1

[0014] Example 1: Acquisition of Phosphoacetylglucosamine Mutase Gene Fragment 1 of White-backed Planthopper

[0015] 1) Using bioinformatics technology to analyze the transcriptome database of the white-backed planthopper, the homology analysis was carried out through the NCBI database Blast program, and a fragment of the SfPAGM gene of the white-backed planthopper was initially screened.

[0016] 2) Based on the sequence of the above-mentioned SfPAGM gene of the white-backed planthopper, the following specific primers were designed using Primer Premier 6.0 software: upstream primer SfPAGM-F 5'-AGATTGTGGAGCCGAGTA-3', downstream primer SfPAGM-R 5'-ATCTTCTTGACGAGGTTGG-3' , all primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd.

[0017] 3) 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....

Embodiment 2

[0020] Example 2: Acquisition of Phosphoacetylglucosamine Mutase Gene Fragment 2 of White-backed Planthopper

[0021] Based on the nucleotide sequence of the white-backed planthopper phosphoacetylglucosamine mutase gene fragment 1 obtained in Example 1, specific primers were designed using Primer Premier 6.0 software, the upstream primer SEQ ID NO: 2, and the downstream primer SEQ ID NO: 3 , all primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. The total RNA was extracted with reference to the method of Example 1, and the extracted RNA was reverse-transcribed into the first-strand cDNA, and PCR amplification was performed to obtain the gene fragment 2 of phosphoacetylglucosamine mutase of white-backed planthopper, Quick Gel Extraction Kit kit for PCR product purification.

Embodiment 3

[0022] Example 3: Synthesis of dsRNA of Phosphoacetylglucosamine Mutase Gene Fragment 2 of White-backed Planthopper

[0023] Phosphoacetylglucosamine mutase gene fragment 2 of white-backed planthopper obtained based on Example 2, using The dsRNA of the SfPAGM gene of the white-backed planthopper was obtained by in vitro transcription, synthesis and purification of the RNAi Kit (Thermo) kit according to the instructions. The purified dsRNA product was subjected to 1% agarose gel electrophoresis to detect its singleness, and a Nanodrop 2000 spectrophotometer (Thermo Fisher) was used to detect its concentration. Store in -80°C refrigerator for later use.

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Abstract

The invention discloses a sogatella furcifera phosphorus acetylglucosamine mutase gene fragment and application thereof. By analyzing a transcriptome database of a sogatella furcifera, a first phosphorus acetylaminoglucose mutase gene fragment is obtained, and the nucleotide sequence of the first phosphorus acetylaminoglucose mutase gene fragment is shown in the sequence table SEQ ID NO:1. A second phosphorus acetylglucosamine mutase gene fragment with the sequence of SEQ ID NO:4 is selected and is used for synthesizing dsRNA of a gene. The dsRNA is transferred to a body of the sogatella furcifera through a microinjection method, and a target gene can be specifically silenced. Abnormal phenotypes of difficult molting or abnormal phenotypes of eclosion are generated during the growth and development of the sogatella furcifera, death is eventually caused, and the death rate reaches 37% after interference for 5 days. The sogatella furcifera phosphorus acetylglucosamine mutase gene fragment provides candidate genes for the future RNA interference mediated pest control strategy of theogatella furcifera, and provides theoretical basis for researching and developing novel pesticides.

Description

technical field [0001] The invention relates to the technical field of insect genetic engineering, in particular to the application of a gene fragment of phosphoacetylglucosamine mutase 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 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 c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/61C12N15/10C12N15/89A01K67/033
CPCA01K67/0339A01K2207/05A01K2217/058A01K2227/706C12N9/90C12N15/10C12N15/1003C12N15/89C12Y504/02003
Inventor 王召朱环王翔金道超杨洪
Owner KAILI UNIV
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