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【目的】果实蝇属Bactrocera中有国际上重要的检疫性害虫,基于形态的物种鉴定有一定的局限性。另一方面,云南边境地区为东南亚地区实蝇入侵我国的重要通道。因此,对该地区实蝇分子鉴定方法的研究对于该属物种的快速准确鉴定具有重要意义。本研究旨在探讨DNA条形码技术在果实蝇属物种鉴定中的有效性。【方法】使用线粒体基因COI和COII序列的通用引物对果实蝇属20个物种60份样品进行PCR扩增、测序和序列分析;采取距离方法和建树方法评价2种序列的鉴别能力。【结果】COI和COII序列平均长度分别为682 bp和339 bp,种内和种间遗传差异较大,有较明显的遗传距离间隔(barcoding gap),鉴定成功率分别为91.2%和90.7%。另外,分子系统树表明华实蝇亚属Sinodacus不是单系群。【结论】COI和COII序列均能够将绝大多数果实蝇属物种进行准确鉴别,应用COI或COII序列进行果实蝇属物种鉴定具有一定的可行性。
【OBJECTIVE】 There are internationally important quarantine pests in fruit fly genus Bactrocera. There are some limitations in the identification of species based on morphology. On the other hand, the Yunnan border area is an important channel for the invasion of fruit flys in Southeast Asia. Therefore, the research on the method of fruit fly identification in this area is of great significance to the rapid and accurate identification of this species. The purpose of this study was to investigate the effectiveness of DNA barcoding in the identification of fruit fly species. 【Method】 The common primers of COI and COII sequences of mitochondrial DNA were used for PCR amplification, sequencing and sequence analysis of 60 samples of 20 genera of fruit flies. The discrimination between the two sequences was evaluated by distance and tree-building methods. 【Result】 The average lengths of COI and COII sequences were 682 bp and 339 bp, respectively. The intraspecific and interspecific genetic differences were large with significant barcoding gap. The successful identification rates were 91.2% and 90.7% respectively. In addition, the molecular phylogenetic tree indicates that Sinodacus subfamily is not a monophyletic group. 【Conclusion】 Both COI and COII sequences can accurately identify most fruit fly species. It is feasible to use COI or COII sequences to identify fruit fly species.