Apolygus lucorum V-ATPaseE gene and application of apolygus lucorum V-ATPaseE gene to aspect of RNAi (ribonucleic acid interfere)-mediated pest control

A pest control and chlorotic bug technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as increased pesticide use, environmental pollution, and chloropylon lack of Bt protein, etc.

Inactive Publication Date: 2017-01-25
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, the prevention and control of Lygus lucidum mainly relies on traditional chemical control due to the lack of effective Bt protein, and the increase in the use of pesticides has seriously polluted the environment.

Method used

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  • Apolygus lucorum V-ATPaseE gene and application of apolygus lucorum V-ATPaseE gene to aspect of RNAi (ribonucleic acid interfere)-mediated pest control
  • Apolygus lucorum V-ATPaseE gene and application of apolygus lucorum V-ATPaseE gene to aspect of RNAi (ribonucleic acid interfere)-mediated pest control
  • Apolygus lucorum V-ATPaseE gene and application of apolygus lucorum V-ATPaseE gene to aspect of RNAi (ribonucleic acid interfere)-mediated pest control

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Embodiment 1

[0025] Example 1 Cloning of ApoV-ATPaseE gene, availability of ApoV-ATPase, dsRNA microinjection and feeding experiment

[0026] Sequencing of the turn green group of A. lucorum and bioinformatics analysis of ApoV-ATPaseE gene.

[0027] Our laboratory performed sequencing of the trans-green group of A. lucorum in advance, and successfully obtained 63029 gene sequences of A. lucorum, and annotated 25,760 gene sequences of them. Using bioinformatics technology and related online software to analyze the transcriptome data of A. lucorum, the ApoV-ATPaseE gene of A. lucorum was determined with the help of online programs such as NCBI Blast.

[0028] The sequencing results are shown in SEQ ID NO:1.

[0029] 2. Collection of test insects and tissues

[0030] L. lucorum was reared using fresh corn (ZeamaysL) and green beans (Phaseolus vulgaris L.) by the Institute of Plant Protection, Chinese Academy of Agricultural Sciences. The rearing temperature is 25-28℃, the relative humidity is 60%-70%,...

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Abstract

The invention discloses an apolygus lucorum V-ATPaseE gene and application of the apolygus lucorum V-ATPaseE gene to the aspect of RNAi (ribonucleic acid interfere)-mediated pest control. Sequences of the apolygus lucorum V-ATPaseE gene are shown as SEQ ID NO:1. Three different segments of dsRNA (double-stranded RNA) are respectively synthesized at different locations of the apolygus lucorum V-ATPaseE gene and are respectively named as dsVA-EF1, dsVA-EF2 and dsVA-EF3, and then the synthesized dsRNA is transmitted to third-instar lavas of apolygus lucorum by the aid of micro-injection methods. The apolygus lucorum V-ATPaseE gene and the application have the advantages that the survival rates of the third-instar lavas are 55%, 70% and 68% respectively in 7 days after the dsRNA is injected into the third-instar lavas as shown by statistics; target genes are obviously silenced as proved by real-time fluorescent quantitative PCR (polymerase chain reaction) results; RNAi feeding experiments are carried out on selected VA-EF1dsRNA, and the survival rates of the third-instar lavas are 65% in 7 days after the dsRNA is injected into the third-instar lavas as shown by results; candidate genes with good effects can be provided for apolygus lucorum RNA interfere (RNAi)-mediated pest control strategies in the future and are verified.

Description

Technical field [0001] The invention relates to the technical field of insect genetic engineering, in particular to a Lygus lucorum V-ATPaseE gene and its application in an RNAi-mediated pest control method. Background technique [0002] In the past decade or so, Bt genetically modified cotton has been planted in more than 90% of the northern provinces of China. The large-scale planting of genetically modified Bt cotton has effectively controlled the cotton bollworm Helicoverpaarmigera (Hubner), the former cotton’s number one enemy, and made the amount of pesticide used in cotton fields Significant economic effects and pest control effects have been obtained by a large reduction. However, the decrease in the amount of pesticide spraying caused the secondary pest Apolygus lucorum (Meyer-Dur) to become more and more rampant. The economic losses caused by A. lucorum in the Yangtze and Yellow River basins have had a great impact on agricultural production every year. So far, the pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/55A01N57/16A01P7/04
CPCC12N9/14A01N57/16
Inventor 王桂荣阿布都如苏力·吐孙刘扬
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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