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Tea geometrid nerve synaptic vesicle protein gene svp and application thereof

A technology of neural synapses and tea inchworms, applied in the fields of application, genetic engineering, plant genetic improvement, etc.

Inactive Publication Date: 2008-07-02
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the use of molecular biology techniques to study the resistance of important pests of Lepidoptera tea plants such as tea loopers has not yet been reported.

Method used

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  • Tea geometrid nerve synaptic vesicle protein gene svp and application thereof

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Effect test

Embodiment 1

[0026]Take 10 3rd-instar tea geometrid larvae with different resistances to kongfuthrin, dilute the commercially available 2.5% kongerin emulsion 10,000 times with water, and then spray it. After 0.5 hours, 3 larvae with obvious poisoning symptoms were selected from the non-resistant larvae group, and 3 larvae with normal performance were selected from the resistant group. The above two types of larvae were ground in different mortars with liquid nitrogen, extracted total RNA with TRIZOL (Invitrogen), and treated with UNIQ-10 Column mRNA Extraction and Purification Kit (Shanghai Sangon Bioengineering Co., Ltd.) to obtain tea geometrid mRNA. Using 1 μg of mRNA as a template, double-stranded cDNA was synthesized using M-MLV RTasecDNA synthesis kit (Bao Bioengineering [Dalian] Co., Ltd.). Add an equal volume of pre-cooled isopropanol to the synthesized double-stranded cDNA, ice-bath for 0.5-12 hours, and then centrifuge at 13,000 rpm for 10-30 minutes. The double-stranded cDNA ...

Embodiment 2

[0032] After diluting the commercially available 2.5% Kungfuthrin emulsion 5000 times with water, the 4th instar larvae of tea geometrid with different pyrethrin pesticide resistance were sprayed. After 2 hours, the larvae sensitive and resistant to Kungfuthrin were taken 1-2 heads (about 0.2g) each, and 1-2 heads of 4th instar larvae that had not been treated with pesticides were taken at the same time. The above samples were placed in a mortar for rapid grinding with liquid nitrogen, and 1ml TRIZOL (Invitrogen Company) was added to mix well. Total RNA was extracted according to the instructions of TRIZOL reagent. According to spectrophotometric analysis, the OD260 / OD280 ratio of the RNA sample was 1.8, and the concentration was about 750 μg / mL. Using 1.5 μL of the above RNA solution as a template, cDNA was synthesized with MMLV first-strand cDNA synthesis kit (Shanghai Sangon Bioengineering Co., Ltd.). Using 1 μL of cDNA as a template and using SEQ ID No: 11 and SEQ ID No: ...

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Abstract

The invention discloses an Ectropis obliqua synaptic vesicle protein gene svp, which has the nucleotide sequence shown in SEQ ID NO: 1. The invention also protein a protein encoded by the gene svp, which has the amino acid sequence shown in SEQ ID NO:2. The invention further discloses the use of the gene fragment svp in evaluation and measurement of the drug resistance of Lepidoptera pests to pesticides.

Description

technical field [0001] The invention belongs to the field of insect genetic engineering. Specifically, the present invention relates to the isolation and analysis of a tea looper synaptic vesicle protein (svp) gene; and also involves using the gene to evaluate the resistance of Lepidoptera pests (tea looper) to pesticides. Background technique [0002] The frequent occurrence of tea geometrid is one of the reasons for the decrease of tea yield and quality. Therefore, tea geometrid is an important object of pest control in tea garden. Pyrethroid pesticides are one of the important pesticides currently used to control tea garden Lepidoptera pests such as tea geometrids. The mode of action of pyrethroid insecticide is contact killing, stomach poisoning and fumigation, but contact killing is the main method; its insecticidal mechanism is to disturb the normal physiology of insect nerves. Due to the characteristics of high efficiency and low toxicity, pyrethroid pesticides are ...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/435C12Q1/68
Inventor 陆建良林晨梁月荣杜颖颖杨晓利
Owner ZHEJIANG UNIV
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