基于16S rDNA与Cytb序列的疏广蜡蝉属三个近似种的分子鉴定(半翅目:蜡蝉总科:广翅蜡蝉科)  

Molecular identification of three similar species of genus Euricania based on 16S rDNA and Cytb gene sequences( Hemiptera: Fulgoroidea:Ricaniidae)

作　　者：刘沅[1,2] 张玉波[1,2,3] 杨琳[1,2] 陈祥盛[1,2,4] 

机构地区：[1]贵州大学昆虫研究所,贵阳550025 [2]昆虫资源开发利用省级特色重点实验室,贵阳550025 [3]安顺学院化学化工学院,贵州安顺561000 [4]贵州大学动物科学学院,贵阳550025

出　　处：《环境昆虫学报》2016年第3期557-564,共8页Journal of Environmental Entomology

基　　金：贵州省国际合作项目(黔科合外G字[2010]7005号);贵州省科技厅三方联合基金(黔科合LH字[2014]7503)

摘　　要：本文通过PCR扩增首次获得了疏广蜡蝉属Euricania Melichar,1898 3个近似种:透明疏广蜡蝉E.clara Kato,1932、长刺疏广蜡蝉E.longa Xu,Liang&Jiang,2006和短刺疏广蜡蝉E.brevicula Xu,Liang&Jiang,2006的16S r DNA序列和Cytb序列;使用MEGA 6.0软件,分析了其序列组成及变异,计算了遗传距离,以宽广蜡蝉属Pochazia Amyot&Serville,1843的圆纹宽广蜡蝉P.guttifera Walker,1851和眼斑宽广蜡蝉P.discreta Melichar,1898为外群,利用邻接法(NJ)和最大简约法(MP)构建了系统发育树。结果显示:透明疏广蜡蝉、长刺疏广蜡蝉和短刺疏广蜡蝉的16S r DNA序列长度分别为414 bp,404 bp,435 bp,而Cytb基因序列长度分别为492 bp,468 bp,472 bp,三者的序列比对、碱基组成成分存在差异;2属5种广翅蜡蝉16S r DNA基因的种间遗传距离为0.008-0.098,属间遗传距离为0.122-0.197,而Cytb基因的种间遗传距离为0.038-0.055,属间遗传距离为0.181-0.188,表明16S r DNA和Cytb基因序列可作为DNA条形码候选基因片段,用于广翅蜡蝉近似属、种的分子鉴定。系统发育树的拓扑结构显示,3个近似种的亲缘关系基本一致,各自聚为一支,且置信值均达96%以上。本文亦结合了透明疏广蜡蝉、长刺疏广蜡蝉和短刺疏广蜡蝉形态学特征的差异对其进行了讨论和分析。16S r DNA and Cytb sequences of three species of Euricania（ Hemiptera： Fulgoroidea：Ricaniidae）,were obtained for the first time. Sequence compositions,variations and genetic distances of the three species were analyzed and the phylogenetic tree of them based on neighbor-joining（ NJ） and maximum parsimony（ MP） methords were constructed using MEGA 6. 0 software,Pochazia guttifera and P. discreta were chosen as outgroups. The results showed that： 16 S r DNA sequence length of E. clara,E.longa and E. brevicula was 414 bp,404 bp,435 bp,and Cytb gene sequence lengths were 492 bp,468 bp,482 bp. Sequence alignment and base composition of them were different; the interspecific genetic distances among these species in 16 S r DNA gene were 0. 008- 0. 098,the genera distances were 0. 122-0. 197,while the interspecific genetic distances among these species in Cytb gene were 0. 038- 0. 055,the genera distances were 0. 181- 0. 188. Therefore as candidate gene fragment of barcoding,16 S r DNA and Cytb genes can be used to molecular identification of species and genera of ricania. The topology of phylogenetic tree indicated that genetic relationship of these species was basically identical,distinguished from each other,and all confidence values reached over 96%. We also discussed and analyzed the differences of morphological characteristics of three euricanian species in this paper.

关 键 词：广翅蜡蝉科 疏广蜡蝉属 16S rDNA CYTB 分子鉴定 

分 类 号：Q963[生物学—昆虫学] S433.39[农业科学—农业昆虫与害虫防治]