董红刚1,谢志娟2,杜予州3,王建军3*,2015,西花蓟马细胞色素P450基因cDNA片段克隆与mRNA表达分析[J].环境昆虫学报,37(6):1188-1194
西花蓟马细胞色素P450基因cDNA片段克隆与mRNA表达分析
Cloning and mRNA expression analysis of cDNA fragments of cytochrome P450 genes in Frankliniella occidentalis
  
DOI:
中文关键词:  西花蓟马  细胞色素P450  RT-PCR  实时荧光定量PCR
英文关键词:Frankliniella occidentalis  cytochrome P450  abamectin resistance  real time PCR
基金项目:公益性行业(农业)科研专项(200803025,201103026)
作者单位
董红刚1,谢志娟2,杜予州3,王建军3* (1. 扬州市邗江区植保植检站江苏扬州 2250092. 泰州市靖江市植保植检站江苏泰州 2145003. 扬州大学园艺与植物保护学院江苏扬州 225009) 
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中文摘要:
      细胞色素P450介导的解毒作用增强是昆虫对杀虫剂产生抗性的重要机制。本文通过RT-PCR克隆了西花蓟马CYP4家族5个细胞色素P450基因cDNA片段。多重序列比对发现,这5个基因在氨基酸水平的一致性在40%-76%之间。采用实时荧光定量PCR对5个CYP4基因的mRNA表达水平的分析发现,阿维菌素抗性品系CYP4-1、CYP4-2和CYP4-5的表达水平分别是敏感品系的3.50、4.00和2.48倍,表明西花蓟马对阿维菌素的抗性可能与这3个CYP4基因的过量表达相关。
英文摘要:
      Enhanced detoxification mediated by cytochrome P450 monoxygenases is a major mechanism of insect resistance to insecticides. In this study, five cDNA fragments of cytochrome P450 genes belonging to the CYP4 family were cloned by RT-PCR from the western flower thrips (WFT), Frankliniella occidentalis. An amino acid sequence alignment shows that these CYP4 genes shared amino acid identity ranging from 40% to 76%. The real-time qPCR analysis of mRNA expression levels of the five CYP4 genes revealed that the relative expression levels of CYP4-1, CYP4-2 and CYP4-5 genes in the abamectin resistant strain were 3.50、4.00 and 2.48 times higher than that in the susceptible strain, respectively, suggesting the possible involvement of overexpression of these three CYP4 genes in the WFT resistance to abamectin.
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