柑橘全爪螨钠离子通道基因差异性分析  

Difference Analysis of Sodium Channel Genes in Panonychus citri(Acari: Tetranychidae)

作　　者：冉春[1] 涂祖霞 陈飞[1,3] 张云飞[1,4] 刘浩强[1] 李鸿筠[1] 

机构地区：[1]中国农业科学院柑桔研究所,国家柑桔工程技术研究中心,重庆400712 [2]重庆市农业学校,重庆401329 [3]西南大学植物保护学院,重庆400716 [4]西南大学园艺园林学院,南方山地园艺学教育部重点实验室,重庆400716

出　　处：《西南大学学报（自然科学版）》2013年第6期1-9,共9页Journal of Southwest University(Natural Science Edition)

基　　金：长江学者和创新团队发展计划项目(IRT0976);国家科技支撑计划课题(2012BAD19B06);柑桔学重庆市市级重点实验室开放基金资助项目

摘　　要：为弄清钠离子通道基因与柑橘全爪螨Panonychus citri噻螨酮抗性的关系,通过柑橘全爪螨转录组分析鉴定了钠离子通道基因;采用序列比对和Sanger测序,分别对柑橘全爪螨敏感品系和噻螨酮抗性品系钠离子通道基因序列进行单核苷酸多态性分析;进一步采用RPKM法和荧光定量PCR对抗性品系和敏感品系钠离子通道基因进行表达差异分析.从柑橘全爪螨转录组中获得了39条钠离子通道基因,相对于敏感品系,抗性品系中有21条表达上调,17条表达下调,Nav1.8.1和Nav1.6.4分别为上调倍数最高的2个钠离子通道基因[log2Ratio(RS/SS)分别为10.59,10.30],进一步荧光定量PCR分析发现,Nav1.8.1和Nav1.6.4上调倍数分别为3.92和1.68;基因序列比较和Sanger测序发现,抗性品系中,Nav1.9.4在344,441,468位和Nav1.7.3在第786分别有3个和1个SNP位点,进一步分析编码的氨基酸序列发现,Nav1.9.4只有第468位G突变为A导致蛋氨酸突变为异亮氨酸,Nav1.7.3在第786位T突变为C导致亮氨酸突变为色氨酸.钠离子通道基因差异性分析及验证为进一步研究柑橘全爪螨抗性分子机理提供了有价值的基因资源.To clarify the relationship between sodium channel genes and hexythiazox-resistance in Panony- chus cirri, sodium channel genes were identified through P. citri transcriptome analysis. SNPs （single nu- cleotide polymorphisms） in sodium channel genes between a susceptible strain and a hexythiazox-resistant strain of P. citri were analyzed by comparison of sodium channel gene sequences and Sanger sequencing. Gene expression profiles of sodium channel genes were further compared, using RPKM （reads per kb per million reads） method and quantitative real-time PCR. Thirty-nine sodium channel transcripts were found in the transcriptome of P. citri. Analysis of differentiated expression of these genes showed that 21 of them were up-regulated and 17 were down-regulated. Navl. 8. 1 and Navl. 6.4 were the two most up-reg- ulated sodium channel genes, their log2 ratio （RS/SS） being 10.59 and 10.30, respectively. Quantitative real-time PCR indicated that Nay1.8.1 and Navl. 6.4 had a 3.92- and 1.68-fold up-regulation, respective- ly, in the resistant strain as compared to the susceptible strain. Comparisons of sodium channel gene se- quences and further Sanger sequencing showed that there were 3 and 1 SNPs in Navl. 9.4 and Navl. 7.3, respectively, between different strains. Analysis of the deduced amino acid sequences revealed that T to C mutation at position 786 in Navl. 7.3 led to a Leu to Ser mutation, and G to A mutation at position 468 in Navl. 9.4 led to a Met to Ile mutation. Difference analysis and validation of sodium channel genes provide a valuable genomic resource for further understanding the molecular basis of the mechanisms of hexythia- zox-resistance in P. citri.

关 键 词：柑橘全爪螨 噻螨酮 钠离子通道基因 差异性分析 

分 类 号：S436[农业科学—农业昆虫与害虫防治]