DNA mini-barcodes in taxonomic assignment: a morphologically unique new homoneurous moth clade from the Indian Himalayas described in Micropterix (Lepidoptera, Micropterigidae)

Abstract

Lees, D. C., Rougerie, R., Zeller-Lukashort, C. & Kristensen, N. P. (2010). DNA minibarcodes in taxonomic assignment: a morphologically unique new homoneurous moth clade from the Indian Himalayas described in Micropterix (Lepidoptera, Micropterigidae).-Zoologica Scripta, 39, 642-661. The first micropterigid moths recorded from the Himalayas, Micropterix cornuella sp. n. and Micropterix longicornuella sp. n. (collected, respectively, in 1935 in the Arunachel Pradesh Province and in 1874 in Darjeeling, both Northeastern India) constitute a new clade, which is unique within the family because of striking specializations of the female postabdomen: tergum VIII ventral plate forming a continuous sclerotized ring, segment IX bearing a pair of strongly sclerotized lateroventral plates, each with a prominent horn-like posterior process. Fore wing vein R unforked, all Rs veins preapical; hind wing devoid of a discrete vein R. The combination of the two first-mentioned vein characters suggests close affinity to the large Palearctic genus Micropterix (to some species of which the members of the new clade bear strong superficial resemblance). Whilst absence of the hind wing R is unknown in that genus, this specialization is not incompatible with the new clade being subordinate within it. A 136-bp fragment of Cytochrome oxidase I successfully amplified from both of the 75-year-old specimens strongly supports this generic assignment. Translated to amino acids, this DNA fragment is highly diagnostic of this genus, being identical to that of most (16 of the 26) Micropterix species studied comparatively here, 1-4 codons different from nine other species (including Micropterix wockei that in phylogenetic analyses we infer to be sister to other examined species), whilst 7-15 codons different to other amphiesmenopteran genera examined here. A dating analysis also suggests that the large clade excluding M. wockei to which M. cornuella belongs appeared <31 million years ago. These findings encourage discovery of a significant radiation of Micropterix in the Himalayan region. Our analysis has more general implications for testing the assignment of DNA mini-barcodes to a taxon, in cases such as museum specimens where the full DNA barcode cannot be recovered.

DNA barcodes from century‐old type specimens using next‐generation sequencing

“…,b; Lees et al . ). Some of these investigations analysed specimens that were relatively young (<50 years), while others extracted DNA from whole specimens.…”

A new species of Micropterix Hübner, 1825 from Lebanon (Lepidoptera: Micropterigidae)

“…The genus Micropterix is distributed through the Palaearctic from North Africa and Europe to Japan in the east (Gibbs 1987;Zeller et al 2013;Gibbs and Lees 2014) and even down to the foothills of the Himalayas (Lees et al 2010). This study of a new species follows the recently published review of Micropterix of Cyprus and the Middle East (Zeller-Lukashort et al 2009).…”

Calibrating the taxonomy of a megadiverse insect family: 3000 DNA barcodes from geometrid type specimens (Lepidoptera, Geometridae)

“…Specimens that failed to generate an amplicon were reanalyzed using primers targeting a 94-bp region and (or) a 64-bp region (Tables 3, 4). (3) Full-length barcodes were recovered from more than 200 century-old specimens using Sanger sequence analysis of multiple short amplicons (Tables 3, 4; for details of the protocol see Lees et al 2010). A NGSbased approach (Prosser et al 2016) was also used to recover full-length barcodes from 101 century-old Geometridae specimens from the NHM (92) and SNSB-ZSM (9), see Fig.…”

“…The first taxonomic revision to include sequence information from an old (150 years) type specimen (Hausmann et al 2009a) involved a timeconsuming and costly "primer walking approach". This protocol did recover the entire DNA barcode through Sanger-sequencing, but it required six PCRs to recover overlapping sequence fragments and 12 sequencing reactions (forward/reverse for each amplicon; see Lees et al 2010 for details on the protocol). Several subsequent studies have used the same approach to recover DNA barcodes from old type specimens of Lepidoptera (Hausmann et al 2009b;Rougerie et al 2012;Strutzenberger et al 2012;Mutanen et al 2015) and other groups (e.g., Puillandre et al 2011).…”