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Apolygus lucorum V-ATPase-A gene cDNA (complementary Deoxyribonucleic Acid) and application thereof

A green Lygus, genetic technology, applied in the field of insect molecular biology and physiology, to achieve a good control effect

Inactive Publication Date: 2017-01-11
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

The development of insect-resistant transgenic crops based on RNAi is currently a research hotspot, and this technology has made some progress in the field of pest control. However, there are few reports on suitable target genes for the green Lygus bug.

Method used

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  • Apolygus lucorum V-ATPase-A gene cDNA (complementary Deoxyribonucleic Acid) and application thereof
  • Apolygus lucorum V-ATPase-A gene cDNA (complementary Deoxyribonucleic Acid) and application thereof
  • Apolygus lucorum V-ATPase-A gene cDNA (complementary Deoxyribonucleic Acid) and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: V-ATPase-A gene cloning, the possession of V-ATPase-A gene dsRNA, microinjection injection of V-ATPase-A gene dsRNA.

[0027] Bioinformatic analysis of genes

[0028] 1. Using bioinformatics technology and software to conduct a preliminary analysis of the transcriptome data of Lygus aeruginosa, and screen out the V-ATPase-A gene of Lygus aeruginosa. The present invention clones the cDNA of V-type ATPase A subunit (V-ATPase-A) from Lygus viridans, which has the sequence shown in SEQ ID NO.1:

[0029] 2. Collection of test insects and tissues

[0030] The plant protection institute of the Chinese Academy of Agricultural Sciences used fresh corn (Zea mays) and green beans (Phaseolus vulgaris L) to raise the green ligus at a temperature of 25-28°C, a relative humidity of 60%-70%, and a light ratio of 16:8. (L:D).

[0031] 3. Extraction of RNA

[0032] a) The whole process of insect total RNA extraction is carried out under RNase-free conditions. The entir...

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Abstract

The invention discloses an apolygus lucorum V-ATPase-A gene cDNA (complementary Deoxyribonucleic Acid) and application thereof, and belongs to the field of biological control. According to the apolygus lucorum V-ATPase-A gene cDNA disclosed by the invention, a V-ATPase-A gene is obtained by cloning apolygus lucorum for the first time; the sequence of the V-ATPase-A gene is shown in SEQ ID NO:1; the gene is used as a target of RNAi (Ribonucleic Acid interference) to design and synthesize apolygus lucorum V-ATPase-AdsRNA; after dsV-ATPase-A is introduced into apolygus lucorum nymphs by adopting an injection method, the V-ATPase-A gene expressions are respectively blocked, the growth and the metabolism of the apolygus lucorum are inhibited, and the death rate is improved. Experiments show that the apolygus lucorum V-ATPase-A gene provides a candidate gene with a good control effect for a future pest control strategy mediated by apolygus lucorum RNAi.

Description

technical field [0001] The invention belongs to the field of insect molecular biology and physiology, and in particular relates to the cloning of the cDNA of the V-ATPase-A subunit (V-ATPase-A) of the green stinkbug, and the preparation of the double-stranded RNA (dsRNA) of the V-ATPase-A gene of the green stinkbug. Design, synthesis and its application in biological control of Lygus pest. Background technique [0002] The green bug (Lygus lucorum), belonging to Hemiptera, Miridae, also known as mosaic bug, small bedbug. Except for a few places such as Qinghai and Tibet, it is distributed in almost all provinces of my country. It is the dominant species of many kinds of Lygus japonica that harm cotton in the Yellow River Basin and Yangtze River Basin of China. Lygus is an omnivorous pest with a wide range of hosts, including cotton, cruciferous vegetables, beans, corn, melons, flowers, potatoes, and various fruit trees, causing huge losses to my country's agriculture and fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/55C12N15/10A01N57/16A01P7/04
CPCC12N9/14A01N57/16C12N15/1096C12Y306/01003
Inventor 王桂荣刘方舟刘杨杨斌
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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