Gwiazdowski Rodger A, Foottit Robert G, Maw H Eric L, Hebert Paul D N
Biodiversity Institute of Ontario, University of Guelph, Guelph, Ontario, N1G 2W1, Canada.
Agriculture and Agri-Food Canada, Invertebrate Biodiversity-National Environmental Health Program, and Canadian National Collection of Insects, Arachnids and Nematodes, Ottawa, Ontario, K1A 0C6, Canada.
PLoS One. 2015 Apr 29;10(4):e0125635. doi: 10.1371/journal.pone.0125635. eCollection 2015.
DNA barcode reference libraries linked to voucher specimens create new opportunities for high-throughput identification and taxonomic re-evaluations. This study provides a DNA barcode library for about 45% of the recognized species of Canadian Hemiptera, and the publically available R workflow used for its generation. The current library is based on the analysis of 20,851 specimens including 1849 species belonging to 628 genera and 64 families. These individuals were assigned to 1867 Barcode Index Numbers (BINs), sequence clusters that often coincide with species recognized through prior taxonomy. Museum collections were a key source for identified specimens, but we also employed high-throughput collection methods that generated large numbers of unidentified specimens. Many of these specimens represented novel BINs that were subsequently identified by taxonomists, adding barcode coverage for additional species. Our analyses based on both approaches includes 94 species not listed in the most recent Canadian checklist, representing a potential 3% increase in the fauna. We discuss the development of our workflow in the context of prior DNA barcode library construction projects, emphasizing the importance of delineating a set of reference specimens to aid investigations in cases of nomenclatural and DNA barcode discordance. The identification for each specimen in the reference set can be annotated on the Barcode of Life Data System (BOLD), allowing experts to highlight questionable identifications; annotations can be added by any registered user of BOLD, and instructions for this are provided.
与凭证标本相关联的DNA条形码参考文库为高通量鉴定和分类学重新评估创造了新机会。本研究为约45%已确认的加拿大半翅目物种提供了一个DNA条形码文库，以及用于生成该文库的公开可用的R工作流程。当前的文库基于对20,851个标本的分析，这些标本包括属于64个科628个属的1849个物种。这些个体被分配到1867个条形码索引号（BINs），即序列簇，它们通常与通过先前分类学认可的物种一致。博物馆藏品是已鉴定标本的关键来源，但我们也采用了高通量采集方法，从而产生了大量未鉴定的标本。这些标本中有许多代表了新的BINs，随后由分类学家进行了鉴定，增加了更多物种的条形码覆盖范围。我们基于这两种方法的分析包括94种未列入最新加拿大清单的物种，这意味着该动物区系可能增加3%。我们在先前DNA条形码文库建设项目的背景下讨论了我们工作流程的发展，强调了划定一组参考标本以帮助解决命名和DNA条形码不一致情况调查的重要性。参考集中每个标本的鉴定可以在生命条形码数据系统（BOLD）上进行注释，以便专家突出有问题的鉴定；任何BOLD注册用户都可以添加注释，并提供了相关说明。
