Building a DNA Barcode Reference Library for the True Butterflies (Lepidoptera) of Peninsula Malaysia: What about the Subspecies?

Abstract

The objective of this study was to build a DNA barcode reference library for the true butterflies of Peninsula Malaysia and assess the value of attaching subspecies names to DNA barcode records. A new DNA barcode library was constructed with butterflies from the Museum of Zoology, University of Malaya collection. The library was analysed in conjunction with publicly available DNA barcodes from other Asia-Pacific localities to test the ability of the DNA barcodes to discriminate species and subspecies. Analyses confirmed the capacity of the new DNA barcode reference library to distinguish the vast majority of species (92%) and revealed that most subspecies possessed unique DNA barcodes (84%). In some cases conspecific subspecies exhibited genetic distances between their DNA barcodes that are typically seen between species, and these were often taxa that have previously been regarded as full species. Subspecies designations as shorthand for geographically and morphologically differentiated groups provide a useful heuristic for assessing how such groups correlate with clustering patterns of DNA barcodes, especially as the number of DNA barcodes per species in reference libraries increases. Our study demonstrates the value in attaching subspecies names to DNA barcode records as they can reveal a history of taxonomic concepts and expose important units of biodiversity.

Can butterflies cope with city life? Butterfly diversity in a young megacity in southern China

“…In a few cases, species pairs sharing DNA barcodes are either very closely related or known to hybridize regularly; consequently, it is not possible to identify them exclusively through DNA barcoding (Dincȃ et al 2011). However, cases of introgressive hybridization have seldom been reported for butterflies (Wilson et al 2013). Furthermore, Smith et al (2012) reported no obvious association between DNA barcode sharing and Wolbachia infection after screening 539 174 DNA barcodes from Lepidoptera (a finding consistent with Linares et al 2009).…”

“…This included species from the families Hesperiidae and Lycaenidae that are difficult to identify using wing morphology (Koh and Sodhi 2004). Although the number of butterfly species in China (1223; Chao 2000) is similar to Peninsular Malaysia (1100; Wilson et al 2013), the number of available DNA barcodes for butterflies from China in BOLD (331) is three times lower than from Peninsular Malaysia (1247). Consequently, most of the DNA barcodes generated for this study were identified based on matches to DNA barcodes from Peninsular Malaysia for which a DNA barcode reference library is available (Wilson et al 2013).…”

“…Although the number of butterfly species in China (1223; Chao 2000) is similar to Peninsular Malaysia (1100; Wilson et al 2013), the number of available DNA barcodes for butterflies from China in BOLD (331) is three times lower than from Peninsular Malaysia (1247). Consequently, most of the DNA barcodes generated for this study were identified based on matches to DNA barcodes from Peninsular Malaysia for which a DNA barcode reference library is available (Wilson et al 2013). This study increased the number of DNA barcodes available in BOLD for butterflies from China five-fold.…”

“…For the vast majority of cases, a priori defined butterfly species can also be delimited unambiguously based on DNA barcodes (Dincȃ et al 2011(Dincȃ et al , 2015Wilson et al 2013). Nevertheless, taxonomic uncertainties during the assembly of reference DNA barcode libraries challenges the use of DNA barcoding for routine species identification (i.e., the assignment of Linnaean species names to unknown specimens) (Collins and Cruickshank 2012).…”

Does infraspecific taxonomy match species evolutionary history? A phylogeographic study of Arundo formosana (Poaceae)

“…This lack of molecular support was also observed among butterflies by Braby, Eastwood & Murray (2012), who called for an integrative concept of subspecies combining phylogenetic support, phenotype distinction and allopatric distribution. In contrast, DNA barcoding of Malaysian butterflies found phylogenetic support for 84% of subspecies (Wilson, Sing & Sofian-Azirun, 2013). The deepest theoretical reflection on infraspecific delimitation in plant taxonomy to date remains the review carried out by Hamilton & Reichard (1992).…”