Cho Soowon, Epstein Samantha W, Mitter Kim, Hamilton Chris A, Plotkin David, Mitter Charles, Kawahara Akito Y
Department of Plant Medicine, Chungbuk National University, Cheongju, South Korea.
Department of Entomology, University of Maryland, College Park, MD, United States.
PeerJ. 2016 Jun 22;4:e2160. doi: 10.7717/peerj.2160. eCollection 2016.
Butterflies and moths (Lepidoptera) comprise significant portions of the world's natural history collections, but a standardized tissue preservation protocol for molecular research is largely lacking. Lepidoptera have traditionally been spread on mounting boards to display wing patterns and colors, which are often important for species identification. Many molecular phylogenetic studies have used legs from pinned specimens as the primary source for DNA in order to preserve a morphological voucher, but the amount of available tissue is often limited. Preserving an entire specimen in a cryogenic freezer is ideal for DNA preservation, but without an easily accessible voucher it can make specimen identification, verification, and morphological work difficult. Here we present a procedure that creates accessible and easily visualized "wing vouchers" of individual Lepidoptera specimens, and preserves the remainder of the insect in a cryogenic freezer for molecular research. Wings are preserved in protective holders so that both dorsal and ventral patterns and colors can be easily viewed without further damage. Our wing vouchering system has been implemented at the University of Maryland (AToL Lep Collection) and the University of Florida (Florida Museum of Natural History, McGuire Center of Lepidoptera and Biodiversity), which are among two of the largest Lepidoptera molecular collections in the world.
蝴蝶和飞蛾（鳞翅目）在世界自然历史收藏中占很大比例，但分子研究缺乏标准化的组织保存方案。传统上，鳞翅目昆虫被展放在标本板上以展示翅膀的图案和颜色，这些对于物种鉴定通常很重要。许多分子系统发育研究使用针插标本的腿部作为DNA的主要来源，以保留形态凭证，但可用组织的量通常有限。将整个标本保存在低温冰箱中对DNA保存来说是理想的，但没有易于获取的凭证会使标本鉴定、核查和形态学研究变得困难。在此，我们介绍一种方法，该方法可为单个鳞翅目昆虫标本创建可获取且易于观察的“翅膀凭证”，并将昆虫的其余部分保存在低温冰箱中用于分子研究。翅膀保存在保护盒中，这样可以轻松查看其背面和腹面的图案和颜色，而不会造成进一步损坏。我们的翅膀凭证系统已在马里兰大学（鳞翅目全生命研究馆藏）和佛罗里达大学（佛罗里达自然历史博物馆，麦圭尔鳞翅目与生物多样性中心）实施，这两个机构拥有世界上最大的两个鳞翅目分子收藏。
