COI及cytb基因对河鲀鱼鱼种鉴定的适用性研究

Application of COI and cytb gene in species identification of pufferfish

目的建立河鲀鱼DNA条形码鉴定方法,探讨细胞色素氧化酶亚基I(COI)及细胞色素b(cytb)基因对我国常见东方鲀属、兔头鲀属河鲀鱼鱼种鉴定的适用性。方法野捕河鲀鱼经形态学鉴定后,钓取COI及cytb基因序列并测序。从Gen Bank下载已有河鲀鱼参考序列,分别构建COI及cytb基因分子进化树,确定样品种属并与形态学鉴定比对。应用所建方法对中毒样品进行河鲀成分鉴定。结果 COI和cytb基因分子进化树将57份样品聚类到东方鲀属和兔头鲀属的9个鱼种,除棕斑兔头鲀和暗鳍兔头鲀(COI进化树)、暗纹东方鲀和晕环东方鲀(cytb进化树)外,2种条形码均能对其余鱼种进行有效区分。中毒样品经鉴定均含有月兔头鲀。结论所建立的DNA条形码方法可有效鉴定河鲀鱼鱼种,弥补形态学鉴定的缺陷。

Objective Develop the identification method for puffer fish species and explorethe applicability of cytochrome oxidase subunit Ⅰ（ COI） and cytochrome b（ cytb） gene in species identification of Takifugu and Lagocephalus. Methods COI and cytb sequences of Takifugu and Lagocephalus were amplified and sequenced. Phylogenetic tree was built by MEGA6. 0 to determine the species of puffer fish. Several poisoning samples of roasted fish fillet were tested by the established method. Results Total 57 samples were classified into 9 species. Phylogenetic tree which were built based on COI and cytb gene could effectively distinguish most species except Lagocephalus spadiceus and Lagocephalus gloveri,Takifugu obscures and Takifugu coronoidus respectively. Lagocephalus lunaris were detected from poisoning samples. Conclusion DNA barcode method could effectively identify puffer fish and make up the defects in morphological identification.