连城国家级自然保护区蝶类DNA条形码库的建立及系统关系分析

Establishment of a butterfly DNA barcoding dataset and phylogenetic analyses based on COI sequence in Liancheng national nature reserve

摘要:

【目的】 构建连城国家级自然保护区蝶类DNA条形码基因库，为蝶类物种快速、准确鉴定提供数据支持。【方法】 于2021-2022年，采用形态和DNA条形码技术对保护区蝴蝶进行种类鉴定，并基于COI序列进行系统发育分析。【结果】 共鉴定保护区蝴蝶5科39属75种，获得145条长度为675 bp的蝶类物种COI序列。其中，塔洒灰蝶（Satyrium thalia）、蓝底霾灰蝶（Maculimea cyanecula）、黄环蛱蝶 （Neptis thems）、黄缘蛱蝶（Ngmphalis antiopa）、西门珍眼蝶（Coenonympha semenovi）、牧女珍眼蝶（C. amaryllis）和古北拟酒眼蝶（Paroeneis palearcticus）的 COI序列均为首次获得。分子鉴定对部分误鉴、 错鉴或难于鉴定的物种进行了有效修订或补充。系统发育分析显示，保护区蝶类在属级水平上系统关系明确，但科级水平上的系统关系尚存在问题。【结论】 连城国家级自然保护区蝶类DNA条形码库的建立不仅有助于蝶类物种的快速准确鉴定，也为DNA条形码技术在物种鉴定中的广泛应用提供了支持；分子鉴定可以对由于人为因素造成的误鉴或错鉴进行校正，是形态学鉴定的良好辅助手段。

Abstract:

【Objective】 The DNA barcode gene bank of butterfly in Liancheng National Nature Reserve was established to provide data support for rapid and accurate identification of butterfly species.【Method】 Systematic investigation of the butterfly resources in the reserve was carried out during 2021-2022. Combination of morphological and DNA barcoding techniques were used for species identification，and a DNA barcoding library of all National Reserve butterfly species was constructed. Then the systematic relationships were analyzed based on the COI sequence. 【Result】 A total of 75 species of butterflies of 5 families and 39 genera were identified in this study，among which 145 specimens were selected for gene extraction and sequencing，and 145 COI sequences of butterfly species with a length of 675 bp were obtained. Among them，Satyrium thalia，Maculimea cyanecula，Neptis thems，Ngmphalis antiopa， Coenonympha semenovi，Coenonympha amaryllis，and Paroeneis palearcticus were obtained for the first time. Comparing the results of morphological and molecular identification，the latter had effectively revised or supplemented some misidentified，misidentified or difficult-to-identify species. The results of phylogenetic analysis based on COI genes showed that the phylogenetic relationships of the protected butterflies were clear at the genus level，but the phylogenetic relationships at the family level were still problematic.【Conclusion】 The establishment of the butterfly DNA barcode database in Liancheng National Nature Reserve not only contributes to the rapid and accurate identification of butterfly species，but also provides support for the wide application of DNA barcode technology in species identification. Molecular identification can correct false identification or misidentification caused by human factors，and is a good auxiliary means of morphological identification.