Biol Invasions (2013) 15:899–910 DOI 10.1007/s10530-012-0338-2 ORIGINAL PAPER Biological invasions in soil: DNA barcoding as a monitoring tool in a multiple taxa survey targeting European earthworms and springtails in North America David Porco • Thibaud Decae¨ns • Louis Deharveng • Samuel W. James • Dariusz Skarzyn_ ´ski • Christer Erse´us • Kevin R. Butt • Benoit Richard • Paul D. N. Hebert Received: 8 October 2011 / Accepted: 31 August 2012 / Published online: 12 September 2012 Ó Springer Science+Business Media B.V. 2012 Abstract Biological invasions are increasingly rec- Collembola and earthworms. Populations of ten spe- ognized as a potent force altering native ecosystems cies of earthworms and five species of Collembola worldwide. Many of the best documented cases were barcoded from both continents. Most of these involve the massive invasions of North America by species exhibited a similar genetic structure of large plant and animal taxa native to Europe. In this study, and stable populations in North America and Europe, a we use DNA barcoding to survey the occurrence and result supporting a scenario of multiple invasions. This genetic structure of two major groups of soil inverte- was expected for earthworm species involved in brates in both their native and introduced ranges: human economic activities, but not foreseen for Collembola species de facto unintentionally intro- duced. This study also establishes that invasive species surveys employing DNA barcoding gain additional Electronic supplementary material The online version of resolution over those based on morphology as they this article (doi:10.1007/s10530-012-0338-2) contains supplementary material, which is available to authorized users. D. Porco (&) Á T. Decae¨ns Á S. W. James Á B. Richard D. Skarzyn_ ´ski Laboratoire d’Ecologie, EA 1293 ECODIV, FED Department of Evolutionary Biology and Ecology, SCALE, Baˆtiment IRESE A, UFR Sciences et Wrocław University, Przybyszewskiego 63/77, Techniques, Universite´ de Rouen, Place Emile Blondel, 51-148 Wrocław, Poland 76821 Mont Saint Aignan, Cedex, France e-mail: [email protected] C. Erse´us Department of Biology and Environmental Sciences, D. Porco Á P. D. N. Hebert University of Gothenburg, Box 463, 405 30 Go¨teborg, Canadian Centre for DNA Barcoding, University Sweden of Guelph, 50 Stone Road East, Guelph, ON N1G 2W1, Canada K. R. Butt School of Built and Natural Environment, University L. Deharveng of Central Lancashire, Preston PR1 2HE, UK Museum National d’Histoire Naturelle, UMR7205 «Origine, Structure et Evolution de la B. Richard Biodiversite´», 45 rue Buffon, CP50, 75005 Paris, France Laboratoire BioSol, Esitpa-Ecole d’Inge´nieurs en Agriculture, 3 rue du Tronquet, 76134 Mont-Saint- Present Address: Aignan, Cedex, France S. W. James Department of Biology, University of Iowa, Iowa City, IA 52242, USA 123 900 D. Porco et al. allow evaluation of cryptic lineages exhibiting differ- horizon mixing, the cycling of carbon, nitrogen ent invasion histories. (Burtelow et al. 1998; Groffman et al. 2004;Sua´rez et al. 2006; Wironen and Moore 2006), phosphorous Keywords Invasion survey Á Underground Á (Sua´rez et al. 2004) and calcium (Holdsworth et al. Molecular taxonomy Á Springtails Á Lumbricidae Á 2008). These changes have major impacts on the plant Anthropic dispersion (Hale et al. 2008), microbial, mesofaunal (Eisenhauer et al. 2007; McLean et al. 2006) and fungal commu- Introduction nities (McLean and Parkinson 2000). By contrast, biological invasions by Collembola Both the pace of non-indigenous species European and their impact on habitats have been much less colonization (Addison 2009), and the success of documented in spite of their ecological significance biological invasions have dramatically increased with (Hopkins 1997). To date, Collembola invasion has the rise of international trade. These biotic exchanges only been documented in a few sub-Antarctic Islands are leading to an increasing homogenization of (Greenslade and Wise 1984; Gabriel et al. 2001; communities (Olden et al. 2004; Winter et al. 2010), Greenslade 2002a; Myburgh et al. 2007; Greenslade and represent one of the main causes of biodiversity 2008; Greenslade and Convey 2012), in Australia decline (Primack 2000; Wilson 2002). Accordingly, (Womersley 1939; King et al. 1985; Greenslade et al. scientific interest in this topic has expanded rapidly 2002; Greenslade 2002b; Oliver et al. 2005; Greens- over the last 30 years. While soil invasions have lade 2008) and in New Zeland (Salmon 1941). Shown generally received little attention, some taxa have to competitively exclude certain native species (Con- recently been studied, e.g. earthworms (Hendrix et al. vey et al. 1999), invasive Collembola display a greater 2008). This neglect is largely due to the ‘taxonomic ability to adapt to rising temperature and so could be impediment’ (Rodman and Cody 2003): in most soil favored by climate changes (Slabber et al. 2007; groups, species identification is a difficult task and Janion et al. 2009). Disturbance, land use changes specialist expertise is often lacking (Decae¨ns et al. (Yeates 1991; Yeates et al. 2000), the use of fertilizers, 2006). In the past, earthworms, among other soil and high grazing intensity (Oliver et al. 2005) also invertebrates, received more attention because they favors invasive species. represent the most conspicuous element of the soil The lack of taxonomic expertise and data on biological invasions due to their large body size and existing invasive taxa in soil biota, emphasize the their role as soil engineers. However, it is undoubt- need for new tools to rapidly assess both local and edly the case that smaller-bodied invaders, having no global situations. In this context, molecular system- such perceivable activity, also occur in soil. In this atic, especially DNA barcoding (Hebert et al. 2003), is study, the invasive patterns in two model taxa of valuable for detecting or identifying invasive species different size are compared: earthworms and Collem- of plants (Van de Wiel et al. 2009) and animals bola from their native (European) and invasive (North (Darling and Blum 2007). Successfully tested in American) ranges. earthworms and Collembola (Rougerie et al. 2009, Over past decades, there has been increasing Decae¨ns et al. 2012), this approach also allows the documentation of the occurrence and consequences identification of juvenile specimens (Richard et al. of the invasions of European earthworms in North 2010; Chown et al. 2008), preventing an underesti- America (Addison 2009). After Pleistocene glacia- mation of the propagule pressure for invasive species. tions, the extinction of native North American earth- This study represents the first multi-taxa applica- worms, except in southern refuges, left many regions tion of DNA barcoding to the investigation of of North America devoid of native species at the time biological invasions in soil. The effectiveness of this of European colonization (Addison 2009), and tool for detection and confirmation of invasive status allowed non-native species spreading (Reynolds in these two target groups is tested. In addition, we aim 1973). Various human activities have led to recurrent to use the DNA barcoding data to compare genetic invasions (Cameron et al. 2007; Cameron et al. 2008). structures between native and invaded areas as a proxy The effects on soil ecosystem properties and func- of invasion history (number of introductions, propa- tioning include major impacts on litter decomposition, gule pressure). 123 Biological invasions in soil 901 Materials and methods Eastern Russia is the main center of diversification for Parisotoma with the highest number of species Sampling (Potapov 2001). Moreover P. notabilis was not found in the most oriental part of the genus’ distribution in In total, 941 specimens belonging to 10 species of Eastern Asia (Potapov 2001; Yosii 1977), while it is Lumbricidae and 275 specimens from 5 species of very abundant in Western Europe. From these choro- Collembola were collected from several localities in logical data, it can be assumed that P. notabilis was Europe and North America (Supplementary Material introduced from Europe to North America; a phylo- Table 1). In addition, 94 sequences of Lumbricidae genetic framework would nevertheless be useful to (AM774234–AM774293, EU035477–EU035492, FJ2 investigate this hypothesis further. 14209–FJ214213, FJ214215, FJ214218, FJ214221, Orchesella villosa and O. cincta were already FJ214222, FJ214227, FJ214230–FJ214232, FJ2142 claimed to be introduced species in North America 34, FJ214235, FJ214237, FJ374776, FJ374777) and (Maynard 1951; Christiansen and Bellinger 1998). one sequence of Collembola (EU869805) were down- Like Parisotoma, Orchesella exhibits its highest loaded from GenBank. All the other specimens were diversity in the old world, with a majority of species processed in the context of the International Barcode found in Europe (Bellinger et al. 2011), by contrast, it of life project (iBOL) for the campaigns ‘Barcoding is only represented in the western regions. So, these Collembola’ and ‘Barcoding Earthworms’ (http:// two species are also likely to originate from Europe. www.earthwormbol.org). Specimen data are avail- Sminthurinus could be another old world genus able in the project ‘DATASET-INVSOIL1’ on the with its highest species diversity in Europe (Bellinger Barcode of Life Data System (www.boldsystems. et al. 2011). This pattern suggests that the species S. org). Each sequence in these libraries