刘沅, 张玉波,杨琳, 陈祥盛*,2016,基于16S rDNA与Cytb序列的疏广蜡蝉属三个近似种的分子鉴定[J].环境昆虫学报,(3):557-564
基于16S rDNA与Cytb序列的疏广蜡蝉属三个近似种的分子鉴定
Molecular identification of three similar species of genus Euricania based on 16S rDNA and Cytb gene sequences
  
DOI:
中文关键词:  广翅蜡蝉科  疏广蜡蝉属  16S rDNA  Cytb  分子鉴定
英文关键词:Ricaniidae  Euricania  16S rDNA  Cytb  molecular identification
基金项目: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)
作者单位
刘沅, 张玉波,杨琳, 陈祥盛* (1. 贵州大学昆虫研究所贵阳 5500252. 昆虫资源开发利用省级特色重点实验室贵阳 5500253. 贵州大学动物科学学院贵阳 5500254. 安顺学院化学化工学院贵州安顺 561000) 
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中文摘要:
      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 rDNA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 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 rDNA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 rDNA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 rDNA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。
英文摘要:
      16S rDNA 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: 16S rDNA 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 16S rDNA 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, 16S rDNA 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.
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