A DNA ‘Barcode Blitz’: Rapid Digitization and Sequencing of a Natural History Collection

DNA barcoding protocols require the linkage of each sequence record to a voucher specimen that has, whenever possible, been authoritatively identified. Natural history collections would seem an ideal resource for barcode library construction, but they have never seen large-scale analysis because of concerns linked to DNA degradation. The present study examines the strength of this barrier, carrying out a comprehensive analysis of moth and butterfly (Lepidoptera) species in the Australian National Insect Collection. Protocols were developed that enabled tissue samples, specimen data, and images to be assembled rapidly. Using these methods, a five-person team processed 41,650 specimens representing 12,699 species in 14 weeks. Subsequent molecular analysis took about six months, reflecting the need for multiple rounds of PCR as sequence recovery was impacted by age, body size, and collection protocols. Despite these variables and the fact that specimens averaged 30.4 years old, barcode records were obtained from 86% of the species. In fact, one or more barcode compliant sequences (>487 bp) were recovered from virtually all species represented by five or more individuals, even when the youngest was 50 years old. By assembling specimen images, distributional data, and DNA barcode sequences on a web-accessible informatics platform, this study has greatly advanced accessibility to information on thousands of species. Moreover, much of the specimen data became publically accessible within days of its acquisition, while most sequence results saw release within three months. As such, this study reveals the speed with which DNA barcode workflows can mobilize biodiversity data, often providing the first web-accessible information for a species. These results further suggest that existing collections can enable the rapid development of a comprehensive DNA barcode library for the most diverse compartment of terrestrial biodiversity – insects.

[1]  M. Smith,et al.  A turbo-taxonomic study of Thai Aleiodes (Aleiodes) and Aleiodes (Arcaleiodes) (Hymenoptera: Braconidae: Rogadinae) based largely on COI barcoded specimens, with rapid descriptions of 179 new species , 2012 .

[2]  C. Mora,et al.  How Many Species Are There on Earth and in the Ocean? , 2011, PLoS biology.

[3]  N. Mills,et al.  DNA Extraction from Museum Specimens of Parasitic Hymenoptera , 2012, PloS one.

[4]  Matthew D Dean,et al.  Factors affecting mitochondrial DNA quality from museum preserved Drosophila simulans , 2001 .

[5]  D. Janzen,et al.  DNA barcodes distinguish species of tropical Lepidoptera. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[6]  Jeremy R. deWaard,et al.  DNA barcodes for 1/1000 of the animal kingdom , 2009, Biology Letters.

[7]  M. Källersjö,et al.  Dichlorvos exposure impedes extraction and amplification of DNA from insects in museum collections , 2010, Frontiers in Zoology.

[8]  L. Keller,et al.  Back to the future: museum specimens in population genetics. , 2007, Trends in ecology & evolution.

[9]  D. Janzen,et al.  Ten species in one: DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[10]  René Tänzler,et al.  One hundred and one new species of Trigonopterus weevils from New Guinea , 2013, ZooKeys.

[11]  A. Hausmann,et al.  DNA Barcoding the Geometrid Fauna of Bavaria (Lepidoptera): Successes, Surprises, and Questions , 2011, PloS one.

[12]  P. Hebert,et al.  The front‐end logistics of DNA barcoding: challenges and prospects , 2009, Molecular ecology resources.

[13]  B. Fontaine,et al.  21 years of shelf life between discovery and description of new species , 2012, Current Biology.

[14]  Michael Bunce,et al.  Highly skewed sex ratios and biased fossil deposition of moa: ancient DNA provides new insight on New Zealand's extinct megafauna , 2010 .

[15]  Jeremy R. deWaard,et al.  An inexpensive, automation-friendly protocol for recovering high-quality DNA , 2006 .

[16]  N. Rohland,et al.  Comparison and optimization of ancient DNA extraction. , 2007, BioTechniques.

[17]  Sujeevan Ratnasingham,et al.  Critical factors for assembling a high volume of DNA barcodes , 2005, Philosophical Transactions of the Royal Society B: Biological Sciences.

[18]  Sujeevan Ratnasingham,et al.  A DNA-Based Registry for All Animal Species: The Barcode Index Number (BIN) System , 2013, PloS one.

[19]  Arturo H. Ariño APPROACHES TO ESTIMATING THE UNIVERSE OF NATURAL HISTORY COLLECTIONS DATA , 2010 .

[20]  M J Scoble,et al.  The web and the structure of taxonomy. , 2007, Systematic biology.

[21]  James Haile,et al.  Non-Destructive Sampling of Ancient Insect DNA , 2009, PloS one.

[22]  I. Guarniero How Many Species Are There on Earth and in the Ocean? (PLOS Biology) , 2014 .