Tuna and tuna-like fishes (tribe Thunnini) are pelagic species that are important worldwide for their ecological and economic value. Understanding the ecology of Thunnini at the early life stages is necessary for proper resource management and conservation. However, the morphological similarity among Thunnini species and the often physically damaged larval and juvenile samples makes unambiguous species identification difficult. Using molecular techniques is one way to deal with these issues, and here we applied high-resolution melting (HRM) analysis for species identification of Thunnini fishes. We developed two species identification sets using HRM analysis based on genotyping species-specific single nucleotide polymorphisms in the mitochondrial genomes of nine species. To validate the effectiveness of the two HRM sets, we screened 312 individuals from 11 Thunnini and one Sardini species. The melting curves from the two sets of HRM analysis showed six perfectly species-specific and three nearly species-specific patterns, with the success rate of species identification ranging from 0.90 to 1.00. Thus, our findings indicate that these two HRM sets are powerful tools that can be used to identify the nine Thunnini species. The ability to identify these species will not only enhance ecological research into their early life stages but also improve the traceability of their fish products.

金枪鱼和类金枪鱼（Thunnini 族）是中上层物种，其生态和经济价值在全世界范围内具有重要意义。了解图尼尼生命早期阶段的生态对于正确的资源管理和保护是必要的。然而，金枪鱼物种之间的形态相似性以及经常受到物理损坏的幼虫和幼鱼样本使得明确的物种鉴定变得困难。使用分子技术是解决这些问题的一种方法，在这里我们应用高分辨率熔化（HRM）分析来鉴定金枪鱼的物种。我们基于九个物种线粒体基因组中物种特异性单核苷酸多态性的基因分型，使用 HRM 分析开发了两个物种鉴定集。为了验证两组 HRM 的有效性，我们筛选了来自 11 个金枪鱼和 1 个沙丁鱼物种的 312 个体。两组HRM分析的熔解曲线显示出6个完全物种特异性的模式和3个接近物种特异性的模式，物种鉴定的成功率在0.90至1.00之间。因此，我们的研究结果表明，这两个 HRM 集是强大的工具，可用于识别九种金枪鱼物种。识别这些物种的能力不仅将加强对其早期生命阶段的生态研究，而且还将提高其鱼产品的可追溯性。我们的研究结果表明，这两个 HRM 集是强大的工具，可用于识别九种金枪鱼物种。识别这些物种的能力不仅将加强对其早期生命阶段的生态研究，而且还将提高其鱼产品的可追溯性。我们的研究结果表明，这两个 HRM 集是强大的工具，可用于识别九种金枪鱼物种。识别这些物种的能力不仅将加强对其早期生命阶段的生态研究，而且还将提高其鱼产品的可追溯性。