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A DNA Barcode Reference Library and Ecoregion Analysis of North American Wolf Spiders (Araneae: Lycosidae)

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A DNA Barcode Reference Library and Ecoregion Analysis of North American Wolf Spiders (Araneae: Lycosidae) Poster ID: 800 A DNA barcode reference library and ecoregion analysis of North American wolf spiders (Araneae: Lycosidae) Gergin A. Blagoev, Jeremy R. deWaard, and Paul D.N. Hebert Centre for Biodiversity Genomics, University of Guelph Abstract Species diversity on our planet follows complex patterns governed by historical, physical and biological parameters, resulting in the classification of well-defined ecoregions usually dominated by specific plant communities. As spiders are predators, they are not directly dependent on plants for nutrition but may have similar distributional patterns. Until now, comprehensive analysis of these patterns has been prohibited by arduous specimen identification and a high incidence of cryptic species. However, DNA barcoding can circumnavigate many of these barriers by delineating species and accelerating specimen identification. We utilize this method to build a DNA barcode reference library for Nearctic wolf spiders (Araneae: Lycosidae), the fifth largest spider family in North America, and investigate distribution patterns across the Nearctic ecoregions. Introduction Although all BIN splits and interim species need further study to clarify the taxonomic status of the entities With 2486 species from 124 genera, the family Lycosidae is involved, DNA barcodes discriminated 85% of all species. ranked fifth in size among spiders (World Spider Catalog, 2017). 70 of the 75 Nearctic World Wildlife Fund ecoregions were Wolf spiders are abundant in almost all terrestrial ecosystems represented by 1-42 BINs (and eleven represented by >20 (Foelix, 2011), and live mainly on the ground surface. This study BINs), which will permit a thorough exploration of distributional develops a reference barcode library for wolf spiders across patterns (Fig. 2A). Canada and the continental United States, which comprises Table 1. Statistical analysis of North American wolf spiders genera 14% of the Earth's surface (Fig. 1) and 14 different ecozones with intraspecific divergence and BIN information, discriminated by (Commission for Environmental Cooperation, 1997). NJ analysis. BIN BIN Min Mean Max Genus n Taxa BINs Methods sharing splits Dist(%) Dist(%) Dist(%) More than 95% of the spider specimens used in this study were Acantholycosa 2 1 1 - - 0 0 0 Allocosa 18 10 11 - 1 5.8 8.4 10.9 collected by members of CBG through various projects in the Alopecosa 615 8 11 2 1 0.7 6.8 10.1 territories of Canada and the United States. However, collections Arctosa 292 10 17 4 5 0 13.6 50 at the Royal BC Museum (Victoria) and the Lyman Entomological Camptocosa 30 2 2 - - 9 9.7 10.3 Museum (Montreal) provided a small proportion of specimens. Geolycosa 227 9 16 - 3 4.1 10.8 17.3 Gladicosa 62 3 3 - - 4.3 5.9 7.6 All specimens were identified or verified by the first author Hogna 304 15 29 - 7 2.7 11.2 17.1 through the genitalic examination of all adults when they were Melocosa 23 1 2 - 1 0 0 0 available. COI sequence analyses were performed at the Pardosa 3800 52 67 22 16 0 5.8 9.9 Pirata 400 15 15 2 - 3 10.4 13.5 Canadian Centre for DNA Barcoding (CCDB; www.ccdb.ca) Piratula 452 4 6 2 1 0.3 7.3 10.9 employing standard protocols outlined in Blagoev et al. (2016). Rabidosa 126 4 6 - 1 5.5 8 9.8 Schizocosa 333 17 29 - 4 3.3 6.7 11.3 Fig. 1. Locality map of all sequenced wolf spiders in the corresponding Sosippus 11 5 5 - - 5.7 9.6 12.7 North American ecozones. Tigrosa 153 5 10 - 1 5.1 9.8 12.9 Trabeops 23 1 1 - - 0 0 0 Trebacosa 7 1 1 - - 0 0 0 Trochosa 373 4 6 - 2 0.2 6.6 12.8 Varacosa 143 3 3 - - 1.9 3.4 4.5 Total 7394 170 241 32 43 Fig. 2. A. Histogram of BIN distribution by number of ecoregions covered; B. Accumulation curves of all valid species names with BINs. BIN Ecoregion Coverage 120 100 80 60 No. BINs 40 Results and Discussion 20 In total, 7394 DNA barcode sequences were generated from 0 A. 1 2 3 4 5 6 7 8 9 10+ B. 170 morphologically-identified species (70% of the Nearctic No. Ecoregions Recorded Lycosidae), representing 241 Barcode Index Numbers (BINs) (Table 1). Adult specimens were not available for 19 BINs, thus Conclusion interim names were assigned. We present a comprehensive DNA barcode reference library for There was a perfect correspondence between BIN membership over two-thirds of the Nearctic wolf spider fauna. The results and a known species in 109 cases, while 32 species shared confirm that DNA barcodes are an effective tool for the BINs, distinguishable by very shallow sequence divergence identification of Nearctic wolf spiders. With the exception of several (<2%). Another 43 species showed deep (>2%) intraspecific cases of potentially cryptic species, BINs and species correspond divergence (i.e. BIN splits). The sequence diversity in wolf well, suggesting that rapid and accurate estimates of spider diversity spider genera shows a clear barcode gap in most cases with a are possible with this approach. It has also permitted the first mean intraspecific divergence of 3.4% vs. a minimum nearest- attempt to resolve the distributional pattern for this large family of neighbour (NN) distance averaging 17.3% (Table 1). spiders in North America. However, if most BIN splits reflect cryptic References taxa (Fig. 2B), the true species count for North American wolf Blagoev, G.A., deWaard, J.R., Ratnasingham, S., deWaard, S.L., Lu, L., Robertson, J., Telfer, A.C. & Hebert, P.D.N., 2016. Untangling taxonomy: a DNA barcode reference library for spiders could be 40–50% higher than currently recognized. Canadian spiders. Molecular Ecology Resources, 16, 325–341. https://doi.org/10.1111/1755- 0998.12444 Acknowledgements Commission for Environmental Cooperation, 1997. Ecological Regions of North America: Financial support was provided by the Ontario Ministry of Research and Innovation and by the Toward a Common Perspective. Montreal, Quebec. 71 pp. government of Canada through Genome Canada and the Ontario Genomics Institute in support Foelix, R., 2011. Biology of Spiders (Third Edition). Oxford University Press. 432 pp. ISBN: of the International Barcode of Life project. We are extremely grateful to colleagues who 9780199734825 provided specimens for analysis (Charles Dondale, Chris Buddle, Claudia Copley, Darren Copley, World Spider Catalog, 2017. World Spider Catalog. Natural History Museum Bern, online at Pat Miller, Robb Bennett, Tom Mason). We are very grateful to John Waithaka and other staff at http://wsc.nmbe.ch, version 18.5, accessed on {date of access}. doi: 10.24436/2. Parks Canada for supporting collection programs in national parks across Canada..
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