几种钝绥螨ITS基因片段的序列分析.pdfVIP

中国农业科学 2011,44(23):4945-4951 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2011.23.023 几种钝绥螨 ITS基因片段的序列分析 邹志文，陈 芬，夏 斌，吴 瑜，张 宇 （南昌大学生命科学与食品工程学院，南昌 330031 ） 摘要：【目的】通过分析比较钝绥螨ITS 基因片段，探讨其作为分子手段应用于种类鉴定，同时应用此分子 标记来分析钝绥螨的亲缘关系，为研究其系统发育和完善分类系统提供参考。【方法】对采自江西桔园的尼氏真 绥螨、江原钝绥螨、津川钝绥螨和东方钝绥螨进行总DNA 提取，PCR 扩增，测序，并从GenBank 中下载5 种钝绥 螨的ITS 序列，应用软件分析ITS 片段长度、A+T%含量、变异位点和进化关系。【结果】得到总长度为644—655 bp 的ITS 序列片段，序列中A+T 碱基比例较高，为58.2%。整个ITS 基因片段存在204 个碱基变异位点（ITS1：172； ITS2：22；5.8S：10）和32 个碱基插入/缺失。真绥螨属的尼氏真绥螨和卵圆真绥螨的分类地位较近，遗传距离 为0.013；而小新绥螨属的4 个种与钝绥螨属的3 个种在构建系统发育树时未能明显地分为2 支，其中胡瓜新小 绥螨和东方钝绥螨的遗传距离最近，为0.083。【结论】ITS 序列片段分析结果支持尼氏真绥螨和卵圆真绥螨现在 的分类地位，而小新绥螨属与钝绥螨属似乎未达到属间差异，其分类地位有待进一步确定。 关键词：钝绥螨；ITS；序列分析 Sequence Analysis of ITS Gene in Several Amblyseiinae (Acari: Phytoseiidae) ZOU Zhi-wen, CHEN Fen, XIA Bin, WU Yu, ZHANG Yu (College of Life Sciences and Food Engineering, Nanchang University, Nanchang 330031) Abstract: 【Objective 】The objective of this study is to analyze the phylogenetic relationships among phytoseiid mites in the citrus orchards by comparing their ITS gene fragments, finally improve the classification system of phytoseiid mites. 【Method 】The fragment sequences of ITS from 4 phytoseiid mites, Euseius nicholsi, Amblyseius eharai , A. tsugawai and A. orientalis , which collected from citrus orchards in Jiangxi province were analyzed by using softwares. 【Result 】Length of ITS from the 4 species are from 644-655 bp. The average A+T content is 58.2%. There are 204 variable sites and 32 insertion/deletion in these sequences. Results demonstrated that ITS2 regions are much more conserved than ITS1. Aligned with the ITS seq