利用DNA条形码技术对茶黑刺粉虱的鉴定与分析

Application of DNA Barcoding Analysis for Identifying Tea Spiny Whitefly(Aleurocanthus camelliae Kanmiya&Kasai)

茶黑刺粉虱(Aleurocanthus camelliae Kanmiya&Kasai)是近年来提出的区别于橘黑刺粉虱[A.spiniferus(Quaintance)]的独立物种,为实现茶黑刺粉虱的精准鉴定,研究利用DNA条形码技术成功获取了来源于贵州独山、贵阳、湄潭以及山东泰安茶园的6个茶黑刺粉虱样本的mt DNA COI基因DNA条形码特定的标准区域序列片段,登录号分别为ON534214、ON534215、ON534319、ON534320、ON548917、ON548918,通过序列比对发现,茶黑刺粉虱个体间的遗传距离(0~8.7%)远小于与外群刺桐粉虱的遗传距离(28.9%~29.8%),证明DNA条形码技术应用有效;系统发育树结果显示,来源于不同地区的茶黑刺粉虱明显分化为2大分支,分别为贵州湄潭、独山和山东泰安分支以及贵州贵阳分支,其结果可支持茶黑刺粉虱的单系性结论,同时2大分支分化明显,尚可能存在隐存种。

In recent publication,the tea spiny whitefly(Aleurocanthus camelliae Kanmiya&Kasai)was considered as a single species distinct from the orange spiny whitefly(A.spiniferus(Quaintance)).To achieve accurate identification to this pest,DNA barcoding sequences of six tea spiny whitefly samples from tea gardens in Meitan,Dushan and Guiyang of Guizhou Province and from Taian of Shandong Province(GenBank accession number:ON534214,ON534215,ON534319,ON534320,ON548917,ON548918)were obtained by DNA extraction and PCR amplification and sequencing of standard DNA barcode fragments of COI gene in this study.Kimura 2-parameter genetic distance analysis showed that intraspecific distances(0-8.7%)of Aleurocanthus camelliae Kanmiya&Kasai was much smaller than interspecific distances from the out-group species Tetraleurodes acaciae(Quaintance)(28.9%-29.8%),proving the effectiveness of the DNA barcoding.Phylogenetic analysis showed that the six samples of tea spiny whitefly were divided into two obvious clades,among which Meitan,Dushan and Taian populations were clustered into one clade,and Guiyang population was clustered into the other clade.These results supported Aleurocanthus camelliae Kanmiya&Kasai to be a monophyletic group;and two clades were differentiated obviously,while potential cryptic species maybe included.