A reference DNA barcode library for Austrian amphibians and reptiles

In the last few years, DNA barcoding became an established method for species identification in biodiversity inventories and monitoring studies. Such studies depend on the access to a comprehensive reference data base, covering all relevant taxa. Here we present a comprehensive DNA barcode inventory of all amphibian and reptile species native to Austria, except for the putatively extinct Vipera ursinii rakosiensis and Lissotriton helveticus, which has been only recently reported for the very western edge of Austria. A total of 194 DNA barcodes were generated in the framework of the Austrian Barcode of Life (ABOL) initiative. Species identification via DNA barcodes was successful for most species, except for the hybridogenetic species complex of water frogs (Pelophylax spp.) and the crested newts (Triturus spp.), in areas of sympatry. However, DNA barcoding also proved powerful in detecting deep conspecific lineages, e.g. within Natrix natrix or the wall lizard (Podarcis muralis), resulting in more than one Barcode Index Number (BIN) per species. Moreover, DNA barcodes revealed the presence of Natrix helvetica, which has been elevated to species level only recently, and genetic signatures of the Italian water frog Pelophylax bergeri in Western Austria for the first time. Comparison to previously published DNA barcoding data of European amphibians and reptiles corroborated the results of the Austrian data but also revealed certain peculiarities, underlining the particular strengths and in the case of the genus Pelophylax also the limitations of DNA barcoding. Consequently, DNA barcoding is not only powerful for species identification of all life stages of most Austrian amphibian and reptile species, but also for the detection of new species, the monitoring of gene flow or the presence of alien populations and/or species. Thus, DNA barcoding and the data generated in this study may serve both scientific and national or even transnational conservation purposes.近年间，DNA条形码（DNA barcoding）已成为生物多样性调查与监测研究中物种鉴定的成熟方法。此类研究依赖于覆盖所有相关类群的全面参考数据库。本研究构建了奥地利本土所有两栖类与爬行类物种的全面DNA条形码名录，仅排除了疑似灭绝的拉科西极北蝰（Vipera ursinii rakosiensis）以及仅在奥地利西部边境有新近报道记录的瑞士真螈（Lissotriton helveticus）。本研究依托奥地利生命条形码（Austrian Barcode of Life, ABOL）计划，共计生成194条DNA条形码序列。通过DNA条形码实现物种鉴定的方法对大多数物种均有效，但在同域分布区域内，杂交起源的水蛙属（Pelophylax spp.）物种复合群以及冠螈属（Triturus spp.）物种除外。然而，DNA条形码技术同样可有效检测到种内深度分化支系，例如在草蛇（Natrix natrix）以及壁蜥（Podarcis muralis）类群中，单个物种可对应多个条形码索引编号（Barcode Index Number, BIN）。此外，本研究通过DNA条形码首次在奥地利西部发现了新近被提升为独立物种的欧洲水游蛇（Natrix helvetica），以及意大利水蛙（Pelophylax bergeri）的遗传特征信号。将本研究数据与已发表的欧洲两栖类、爬行类DNA条形码数据进行比对后，不仅验证了奥地利本土数据集的分析结果，同时也揭示了部分特异性特征，既凸显了DNA条形码技术的独特优势，也体现了其在侧褶蛙属（Pelophylax）类群应用中的局限性。因此，DNA条形码技术不仅可有效鉴定奥地利大多数两栖类与爬行类物种各生命阶段的个体，还可用于新物种发现、基因流监测以及外来种群/物种的入侵检测。综上，DNA条形码技术及本研究生成的数据集，可同时服务于科学研究以及国家级乃至跨国层面的生物多样性保护工作。