Abstract
DNA barcoding, based on a fragment of cytochrome c oxidase I (COI) mtDNA, is as an effective molecular tool for identification, discovery, and biodiversity assessment for most animals. However, multiple gene markers coupled with more sophisticated analytical approaches may be necessary to clarify species boundaries in cases of cryptic diversity or morphological plasticity. Using 339 moths collected from mountains surrounding Beijing, China, we tested a pipeline consisting of two steps: (1) rapid morphospecies sorting and screening of the investigated fauna with standard COI barcoding approaches; (2) additional analyses with multiple molecular markers for those specimens whose morphospecies and COI barcode grouping were incongruent. In step 1, 124 morphospecies were delimited into 116 barcode units, with 90% of the conflicts being associated with specimens identified to the genus Hypena. In step 2, 55 individuals representing all 12 Hypena morphospecies were analysed using COI, COII, 28S, EF-1a, Wgl sequences or their combinations with the BPP (Bayesian Phylogenetics and Phylogeography) multigene species delimitation method. The multigene analyses supported the delimitation of 5 species, consistent with the COI analysis. We conclude that a two-step barcoding analysis pipeline is able to rapidly characterize insect biodiversity and help to elucidate species boundaries for taxonomic complexes without jeopardizing overall project efficiency by substantially increasing analytical costs.
Highlights
DNA barcoding – the sequencing of a short, standard genetic marker from unknown specimens coupled with analyses of sequence divergences1,2 – has been shown to be a practical tool for species identification and biodiversity assessment[3,4,5]
A mitochondrial single-locus approach can occasionally be problematic for accurate species delimitation[27,28] and, especially for taxonomically contentious groups, “independent” nuclear genes may be needed as supplementary markers to support any conclusions[11]
The Bayesian Phylogenetics and Phylogeography (BPP) method is grounded on the multispecies coalescent model and calculates the posterior probabilities of competing models that contain more, or fewer, lineages
Summary
DNA barcoding – the sequencing of a short, standard genetic marker from unknown specimens coupled with analyses of sequence divergences1,2 – has been shown to be a practical tool for species identification and biodiversity assessment[3,4,5]. Brower[14] concluded that there were only three to seven cryptic species of Astraptes fulgerator rather than the ten suggested by Hebert et al.[8], while Dasmahapatra et al.[15] concluded that only one of four ‘cryptic species’ in the butterfly genus Mechanitis was biologically meaningful Despite these disagreements, which may reflect disparities in notions of what constitutes a species and how they are recognized, a general consensus has emerged that standard COI barcodes can meet the needs of much conventional species identification and delimitation[2,16,17,18,19,20,21,22]. The BPP method is grounded on the multispecies coalescent model and calculates the posterior probabilities of competing models that contain more, or fewer, lineages
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