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Lepidoptera transposon gene tpa and application thereof

A lepidopteran transposon and lepidopteran technology, which is applied to the isolation of lepidopteran pest transposon gene fragments, and the application field of tpa gene in the detection of lepidopteran pest resistance, which can solve the problem of drug resistance and pest resistance. The genetic mechanism of medicinal properties needs to be studied, and the evaluation indicators of pesticide resistance are not clear.

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

AI Technical Summary

Problems solved by technology

However, due to years of continuous use, lepidopteran pests have become more and more tolerant to pyrethroid pesticides, and some even have developed resistance
Pesticide resistance evaluation indicators of pests are not yet clear, and the genetic mechanism of pest resistance remains to be studied

Method used

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  • Lepidoptera transposon gene tpa and application thereof

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Embodiment 1

[0022] Take 30 3rd-instar tea geometrid larvae with moderate resistance and strong resistance to kungfuthrin, and another 30 3rd-instar tea geometrid larvae without drug resistance, and raise them in three 2L beakers respectively, and feed them with tea leaves normally. After the new shoots (5-8 / cup), the commercially available concentration of 2.5% kaufthrin emulsion is diluted 5000 times with water and then sprayed. 20 minutes after spraying, select 2-3 larvae from different drug-resistant larvae. The above larval samples were ground in different mortars with liquid nitrogen, total RNA was extracted with TRIZOL (Invitrogen) reagent, and mRNA was isolated with UNIQ-10 column mRNA extraction and purification kit (Shanghai Sangon Bioengineering Co., Ltd.). 1 μg of mRNA was used for cDNA synthesis, and double-stranded cDNA was synthesized using TakaRa MMLV RTase cDNA Synthesis Kit (Takara Bioengineering [Dalian] Co., Ltd.). After the double-stranded cDNA was purified by isoprop...

Embodiment 2

[0025]40 tea geometrids and tea caterpillar 3rd instar larvae with different resistance to kungfuthrin were taken, and each pest was divided into two groups according to their resistance, namely sensitive group and resistant group. The commercially available concentration of 2.5% kaufthrin emulsion is diluted 10000 times with water and then sprayed. After 2 hours, 1-2 larvae were selected from each of the sensitive and resistant groups for tpa expression studies. RNA was extracted by TRIZOL (Invitrogen Company), and cDNA was synthesized by MMLV first-strand cDNA synthesis kit (Shanghai Sangon Bioengineering Co., Ltd.). Using 1 μL of cDNA as a template, PCR was performed using primer SEQ ID No: 10 (since the target sequence is an inverted repeat sequence, only one primer is needed, and the product length is 434 bp). After PCR, constant voltage electrophoresis was performed on 1.5% agarose gel, and photographs were taken after electrophoresis, as shown in Figure 2. It can be s...

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Abstract

The invention discloses a Lepidoptera transposon gene fragment tpa, which has the nucleotide sequence shown in SEQ ID NO:1. The invention also discloses the use of the gene fragment tpa 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, and specifically relates to the isolation of transposase (tpa) gene fragments of lepidopteran pests, and also relates to the application of the tpa gene in the detection of drug resistance of lepidopteran pests. Background technique [0002] Lepidoptera pests often cause tea yield reduction and quality decline, so they are the main pest control targets in tea gardens, and pyrethroids are commonly used pesticides to control Lepidoptera pests. However, due to years of continuous use, lepidopteran pests have become more and more tolerant to pyrethroid pesticides, and some have even developed resistance. Pesticide resistance evaluation indicators of pests are not yet clear, and the genetic mechanism of pest resistance remains to be studied. Contents of the invention [0003] The technical problem to be solved by the present invention is to provide a lepidopteran transposon gene fragment t...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12Q1/68
Inventor 陆建良梁月荣林晨郑新强董俊杰董占波
Owner ZHEJIANG UNIV
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