The use of molecular tools (DNA barcoding and metabarcoding) for the identification of species and ecosystem biomonitoring is a promising innovative approach. The effectiveness of these tools is, however, highly dependent on the reliability and coverage of the DNA sequence reference libraries and it also depends on the identification of primer sets that work on the broadest range of taxa. In this study, a gap analysis of available DNA barcodes in the international libraries was conducted using the aquatic macroinvertebrate species checklist of the Apulia region in the southeast of Italy. Our analyses show that 42% of the 1546 examined species do not have representative DNA barcodes in the reference libraries, indicating the importance of working toward their completeness and addressing this effort toward specific taxonomic groups. We also analyzed the DNA barcode reference libraries for the primer set used to barcode species. Only for 52% of the examined barcoded species were the primers reported, indicating the importance of uploading this information in the databases for a more effective DNA barcode implementation effort and extensive use of the metabarcoding method. In this paper, a new combination of primers has revealed its experimental effectiveness at least on the species belonging to the three most represented taxa in the aquatic ecosystems of the Apulia region, highlighting the opportunity to develop combinations of primers useful at the regional level and the importance of studying DNA barcode gaps at the local/regional level. The DNA barcode coverage also varies among different taxonomic groups and aquatic ecosystem types in which a large number of species are rare. We tested the application of the DNA barcoding single species to a lagoon ecosystem (the lagoon named “Acquatina di Frigole” in the Apulia region) and we sampled two macroinvertebrate species lacking DNA barcodes from “Aquatina di Frigole” NATURA 2000 Site IT9150003, Fabulina fabula and Tritia nitida, generated two new CO1 barcodes and added them to a DNA barcode reference library.

中文翻译：

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普利亚地区（意大利东南部）水生大型无脊椎动物物种DNA条形码参考文库的差距分析

使用分子工具（DNA条形码和元条形码）来识别物种和进行生态系统生物监测是一种很有前途的创新方法。但是，这些工具的有效性高度依赖于DNA序列参考文库的可靠性和覆盖范围，并且还取决于鉴定适用于最广泛分类单元的引物组。在这项研究中，使用意大利东南部普利亚地区的水生无脊椎动物物种清单对国际图书馆中可用的DNA条码进行了缺口分析。我们的分析表明，在1546个受检查物种中，有42％在参考文库中没有代表性的DNA条码，这表明了提高其完整性和针对特定分类组的努力的重要性。我们还分析了DNA条形码参考文库中用于条形码种类的引物集。仅报告了52％的条形码条形码种类，表明将这些信息上载到数据库中对于更有效的DNA条形码实施工作和广泛使用元条形码方法的重要性。在本文中，一种新的引物组合显示了其至少对属于普利亚地区水生生态系统中三个代表性最强的分类单元的物种具有实验有效性，这突出显示了开发在区域一级使用的引物组合的机会。在本地/区域级别研究DNA条形码缺口的重要性。DNA条形码的覆盖率在不同的生物分类群和水生生态系统类型之间也有所不同，在这些种类中，稀有物种很少。我们测试了将DNA条形码的单个物种应用于泻湖生态系统（在普利亚地区的泻湖命名为“ Acquatina di Frigole”），并从NATURA 2000 Site IT9150003的“ Aquatina di Frigole”采样了两个缺少DNA条形码的大型无脊椎动物。Fabulina fabula和Tritia nitida产生了两个新的CO1条形码并将其添加到DNA条形码参考库中。