Abstract Accurate species identification is crucial in ecological research on sentinel species such as earthworms. Here, we developed a reliable and cost-efficient method for the rapid identification of taxa within the Allolobophora chlorotica complex of cryptic earthworm species. We combined high resolution melting (HRM) analysis with DNA barcoding (Bar-HRM) in a three-step approach: (i) selection of a mini-barcode in the 16S mitochondrial gene, (ii) test of the method on a panel of 16 reference individuals and (iii) identification at the species level of 24 unknown specimens. Thus, we proved the efficacy of Bar-HRM to discriminate between cryptic sister (or sibling) earthworm species and we provided a standardized method that can be easily adapted to other taxa.

中文翻译：

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Bar-HRM 用于鉴定隐匿的蚯蚓物种

摘要 准确的物种识别是蚯蚓等哨兵物种生态研究的关键。在这里，我们开发了一种可靠且具有成本效益的方法，用于快速识别隐匿蚯蚓物种的 Allolobophora chlorotica 复合体中的分类群。我们以三步法将高分辨率熔解 (HRM) 分析与 DNA 条形码 (Bar-HRM) 相结合：(i) 在 16S 线粒体基因中选择迷你条形码，(ii) 在一组16 个参考个体和 (iii) 在物种水平鉴定 24 个未知标本。因此，我们证明了 Bar-HRM 在区分神秘姐妹（或兄弟）蚯蚓物种方面的功效，并且我们提供了一种可以轻松适应其他分类群的标准化方法。