DNA条形码技术在田间常见蓟马种类识别中的应用  

Application of DNA barcoding technology for species identification of common thrips(Insecta:Thysanoptera) in China

作　　者：乔玮娜[1] 万方浩[1,2] 张爱兵[3] 闵亮[1] 张桂芬[1,2] 

机构地区：[1]中国农业科学院植物保护研究所,植物病虫害生物学国家重点实验室,北京100193 [2]农业部外来入侵生物预防与控制研究中心,北京100081 [3]首都师范大学生命科学学院,北京100048

出　　处：《昆虫学报》2012年第3期344-356,共13页Acta Entomologica Sinica

基　　金：国家重点基础研究发展规划(“973”计划)项目(2009CB119200);国家公益性行业(农业)科研专项(201103026);国家自然科学基金项目(30971967)

摘　　要：蓟马类害虫种类多、体型小,传统的形态学鉴定方法难以快速准确识别。本研究利用DNA条形码通用型引物,以我国田间常见的25种蓟马为靶标扩增其线粒体DNA细胞色素C氧化酶亚基Ⅰ(mitochondrial cytochrome coxidase subunit Ⅰ gene,mtDNA COⅠ)基因(约650bp),通过对靶标片段碱基序列的测序及比对分析,以邻接法(NJ法)构建系统发育树,并以Kimura双参数模型计算种内、种间遗传距离。结果表明:聚类分析与形态学鉴定结果一致,表现为较长的种间分支和较短的种内分支,每个单系分支对应一个物种,同一物种不同单倍型的最初分支自展值均为100%。25种蓟马的种内平均遗传距离为0.0027,种间平均遗传距离为0.2757,种间遗传距离为种内遗传距离的102.1倍;而且种内、种间遗传距离没有重叠区域。结果说明基于COⅠ基因的DNA条形码技术可以用于不同种类蓟马的快速准确鉴别。Thrips（Insecta：Thysanoptera） are of a species-rich order with small size,which makes them hard to be identified quickly and accurately by traditional morphological identification methods.In this study,the mitochondrial cytochrome c oxidase subunit Ⅰ（COⅠ） genes of 25 thrips species common in China were amplified using DNA barcoding universal primers.The obtained partial COⅠ fragments（about 650 bp） were sequenced.The phylogenetic tree was established by a neighbor-joining（NJ） method.The intra-and inter-species genetic distances were calculated with MEGA version 4.0 software using the Kimura 2-Parameter model.The results indicated that the identification results based on the phylogenetic tree obtained was consistent with those based on morphological characteristics for the 25 thrips species.In these species,the lengths of intra-species branches were much shorter than those of the inter-species branches.Each branch corresponded to one species and all bootstrap values of the original divergence among different haplotypes within one species were 100%.The genetic distance between species for these 25 species was 102.1 times as high as the corresponding values within species（0.2757 vs.0.0027）.There was no overlap between inter-and intra-species genetic distances.The results suggest that the DNA barcoding based on partial COⅠ gene can provide rapid and accurate identification of these thrips species.

关 键 词：缨翅目 蓟马 COⅠ基因 DNA条形码 分子鉴定 遗传距离 系统发育树 

分 类 号：Q966[生物学—昆虫学]