慈竹叶蝉类害虫DNA条形码分析
DNA Barcoding of Bambusa emeiensis-Feeding Leafhoppers in China for Species Identification

中文摘要：叶蝉类昆虫形态结构多样，在农林生态系统的物种多样性和植物保护工作中扮演着重要角色，但其物种的准确鉴定一直是农林植保工作中的难点。DNA条形码技术极大促进了农林生态系统物种的快速、准确鉴定。本研究经过连续2年的野外调查采集慈竹 Bambusa emeiensis主要叶蝉种类，扩增了广泛分布于中国慈竹的12种主要叶蝉类害虫的线粒体基因 COⅠ和 16S rRNA序列片段，并进行了遗传距离、系统发育及矢量Klee-diagram图分析。结果显示：慈竹叶蝉昆虫 COⅠ基因序列片段（590 bp）种内遗传距离为0.004，种间遗传距离为0.283； 16S rRNA基因序列片段（463 bp）种内遗传距离为0.003，种间遗传距离为0.257；不同种间存在明显的条形码间隔。2个基因序列片段的分子系统发育分析结果与形态学研究谱系关系一致。Klee-diagram图分析结果和分子系统发育结果一致。上述结果表明， COⅠ和 16S rRNA基因适用于慈竹叶蝉类昆虫的物种鉴定，可为竹林叶蝉类昆虫的准确快速鉴定提供参考方法。
英文摘要：Leafhoppers play a key role in biodiversity and plant protection of agricultural and forestry ecosystems. However, species identification of leafhoppers is a challenge for this subject due to their morphology diversity. DNA barcoding provides an efficient method for species identification. In the present study, partial COⅠ and 16S rRNA gene sequences from 12 Bambusa emeiesis-feeding leafhopper species in China were amplified. A neighbor-joining phylogenetic tree was constructed based on Kimura 2-parameter (K2P) genetic distance, and further studied using Klee-diagram vector analysis. Intra-specific K2P genetic distance averaged 0.004 for COⅠ, and 0.003 for 16S rRNA; inter-specific K2P genetic distance averaged 0.283 and 0.257 for COⅠ and 16S rRNA, respectively. An obvious ‘barcoding gap’ was detected for all species in both markers, and all individuals were clustered together in both phylogeny trees constructed using COⅠ and 16S rRNA. The result of cluster analyses based on COⅠ and 16S rRNA gene sequences were basically consistent with that of morphological identification, and different individuals tested in this study were gathered in a common branch of the dendrograms. Moreover, the result of Klee-diagram vector analysis was also consistent with phylogeny. Hence, both COⅠ and 16S rRNA genes appear to be accurate and efficient markers for bamboo leafhoppers identification in China. 2018,37(2): 127-138 收稿日期：2017-09-23 DOI：10.11984/j.issn.1000-7083.20170294 分类号：Q953;Q969.35 基金项目：国家自然科学基金项目（31260178，31660209）；贵州省高层次创新型人才（"百"层次）项目（黔科合人才20154021）；贵州省节肢动物资源开发利用科技创新人才团队项目（黔科合人才团队[2014]4001号） 作者简介：姚亚林(1990-),男,硕士研究生,主要从事昆虫区系分类和农林昆虫物种多样性研究,E-mail:yaoyalin08@sina.com *通讯作者：杨琳,E-mail:yanglin6626@163.com 参考文献： Del-Prado R, Cubas P, Lumbsch HT, et al. 2010. Genetic distances within and among species in monophyletic lineages of Parmeliceae (Ascomycota) as a tool for taxon delimitation[J]. Molecular Phylogenetics and Evolution, 56:125-133. Dietrich CH, Whitcomb RF, Black IV WC. 1997. Phylogeny of the grassland leafhopper genus Flexamia (Homoptera:Cicadellidae) based on mitochondrial DNA