Invertebrate Systematics Invertebrate Systematics Society
Systematics, phylogeny and biogeography
RESEARCH ARTICLE

Challenging species delimitation in Collembola: cryptic diversity among common springtails unveiled by DNA barcoding

D. Porco A H , A. Bedos B , Penelope Greenslade C , C. Janion D , D. Skarżyński E , M. I. Stevens F , B. Jansen van Vuuren G and L. Deharveng B

A Université de Rouen – Laboratoire ECODIV, Bâtiment IRESE A, Place Emile Blondel, 76821 Mont Saint Aignan Cedex, France.

B Muséum National d’Histoire Naturelle, UMR7205 «Origine, Structure et Evolution de la Biodiversité», 45 rue Buffon, CP50, 75005 Paris, France.

C Environmental Management, School of Science, Information Technology and Engineering, University of Ballarat, Ballarat, Vic. 3353, Australia.

D Centre for Invasion Biology, Department of Botany and Zoology, Stellenbosch University, Private Bag x1, Matieland 7602, South Africa.

E Zoological Institute, Wrocław University, Przybyszewskiego 63/77, 51-148 Wrocław, Poland.

F South Australian Museum, GPO Box 234, Adelaide, SA 5000, and School of Earth and Environmental Sciences, University of Adelaide, SA 5005, Australia.

G Centre for Invasion Biology, Department of Zoology, University of Johannesburg, PO Box 524, Auckland Park 2006, South Africa.

H Corresponding author: Email: david.porco.gm@gmail.com

Invertebrate Systematics 26(6) 470-477 http://dx.doi.org/10.1071/IS12026
Submitted: 15 April 2012  Accepted: 22 September 2012   Published: 19 December 2012

Abstract

Collembola is one of the major functional groups in soil as well as a model taxon in numerous disciplines. Therefore the accurate identification of specimens is critical, but could be jeopardised by cases of cryptic diversity. Several populations of six well characterised species of springtails were sequenced using the COI barcode fragment as a contribution to the global Collembola barcoding campaign. Each species showed high intraspecific divergence, comparable to interspecific sequence divergence values observed in previous studies and in 10 congeneric species barcoded here as a reference. The nuclear marker, 28S, confirmed all the intraspecific lineages found with COI, supporting the potential specific status of these entities. The implications of this finding for taxonomy and for disciplines relying on species names, such as evolution and ecology, are discussed.


References

Barateiro Diogo, J., Natal-Da-Luz, T., Paulo Sousa, J., Vogt, C., and Nowak, C. (2007). Tolerance of genetically characterized Folsomia candida strains to phenmedipham exposure – a comparison between reproduction and avoidance tests. Journal of Soils and Sediments 7, 388–392. open url image1

Busmachiu, G., Deharveng, L., and Weiner, W. M. (2010). A new species of the genus Lathriopyga Caroli, 1912 (Collembola, Neanuridae, Neanurinae) from the Republic of Moldova. Zootaxa 2639, 53–58. open url image1

Carapelli, A., Fanciulli, P. P., Frati, F., and Dallai, R. (1995a). The use of genetic markers for the diagnosis of sibling species in the genus Isotomurus (Insecta, Collembola). Bollettino di Zoologia 62, 71–76.
The use of genetic markers for the diagnosis of sibling species in the genus Isotomurus (Insecta, Collembola).CrossRef | open url image1

Carapelli, A., Frati, F., Fanciulli, P. P., and Dallai, R. (1995b). Genetic differentiation of six sympatric species of Isotomurus (Collembola, Isotomidae) – is there any difference in their microhabitat preference? European Journal of Soil Biology 31, 87–99. open url image1

Cassagnau, P. (1976). Variability of polytene chromosomes in Bilobella aurantiaca Caroli (Collembola, Neanuridae) its relations with biogeography and ecology of species. Archives De Zoologie Experimentale Et Generale 117, 511–572. open url image1

Cassagnau, P. (1991). Les collemboles Neanurinae de l’Himalaya 2: Paranurini et Paleonurini paucitubercules. Travaux du Laboratoire d’Ecobiologie des Arthropodes Edaphiques Toulouse 6, 1–20. open url image1

Cassagnau, P., Dallai, R., and Deharveng, L. (1979). Polytene chromosomes polymorphism in Lathriopyga longiseta Caroli (Collembola Neanuridae). Caryologia 32, 461–483. open url image1

Chenon, P., Rousset, A., and Crouau, Y. (2000). Genetic polymorphism in nine clones of a parthenogenetic collembolan used in ecotoxicological testing. Applied Soil Ecology 14, 103–110.
Genetic polymorphism in nine clones of a parthenogenetic collembolan used in ecotoxicological testing.CrossRef | open url image1

Cicconardi, F., Nardi, F., Emerson, B. C., Frati, F., and Fanciulli, P. P. (2010). Deep phylogeographic divisions and long-term persistence of forest invertebrates (Hexapoda: Collembola) in the North-Western Mediterranean basin. Molecular Ecology 19, 386–400.
Deep phylogeographic divisions and long-term persistence of forest invertebrates (Hexapoda: Collembola) in the North-Western Mediterranean basin.CrossRef | 1:CAS:528:DC%2BC3cXktlOntLc%3D&md5=1b3a8a76f62f1500e24a9776dd9275deCAS | open url image1

Crouau, Y., Cazes, L., and Bur, T. (2009). The use of the Collembola reproduction test in bioassays of polluted soils. In ‘Progress in Environmental Science and Technology. Vol II, Pts A and B’. (Eds S. C. Li, Y. J. Wang, F. X. Cao, P. Huang and Y. Zhang.) pp. 1843–1845. (Science Press Beijing: Beijing.)

D’Haese, C. A. (2002). Were the first springtails semi-aquatic? A phylogenetic approach by means of 28S rDNA and optimization alignment. Proceedings. Biological Sciences 269, 1143–1151.
Were the first springtails semi-aquatic? A phylogenetic approach by means of 28S rDNA and optimization alignment.CrossRef | 1:CAS:528:DC%2BD38Xls1aqt7w%3D&md5=c7513daf065cf5c812152f95dcb49a38CAS | open url image1

Deharveng, L. (2004). Recent advances in Collembola systematics. Pedobiologia 48, 415–433.
Recent advances in Collembola systematics.CrossRef | open url image1

Fanciulli, P. P., Frati, F., Dallai, R., and Rusek, J. (1991). High genetic divergence among populations of Tetrodontophora bielanensis (Insecta, Collembola) in Europe. Revue D Ecologie Et De Biologie Du Sol 28, 165–173. open url image1

Fanciulli, P. P., Melegari, D., Carapelli, A., Frati, F., and Dallai, R. (2000). Population structure, gene flow and evolutionary relationships in four species of the genera Tomocerus and Pogonognathellus (Collembola, Tomoceridae). Biological Journal of the Linnean Society. Linnean Society of London 70, 221–238. open url image1

Fanciulli, P. P., Carapelli, A., Belloni, M., Dallai, R., and Frati, F. (2009). Allozyme variation in the springtails Allacma fusca and A. gallica (Collembola, Sminthuridae). Pedobiologia 52, 309–324.
Allozyme variation in the springtails Allacma fusca and A. gallica (Collembola, Sminthuridae).CrossRef | open url image1

Folmer, O., Black, M., Hoeh, W., Lutz, R., and Vrijenhoek, R. (1994). DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology 3, 294–299.
| 1:CAS:528:DyaK2MXjt12gtLs%3D&md5=0a6e1d841a9ef8fafdf936427c59141bCAS | open url image1

Frati, F., Fanciulli, P. P., and Dallai, R. (1992). Genetic diversity and taxonomy in soil-dwelling insects – the genus Orchesella. The Journal of Heredity 83, 275–281. open url image1

Frati, F., Carapelli, A., Fanciulli, P. P., and Dallai, R. (1995). The genus Isotomurus: where molecular markers help to evaluate the importance of morphological characters for the diagnosis of species. Polskie Pismo Entomologiczne 64, 41–51. open url image1

Frati, F., Negri, I., Fanciulli, P. P., Pellecchia, M., De Paola, V., Scali, V., and Dallai, R. (2004). High levels of genetic differentiation between Wolbachia-infected and non-infected populations of Folsomia candida (Collembola, Isotomidae). Pedobiologia 48, 461–468.
High levels of genetic differentiation between Wolbachia-infected and non-infected populations of Folsomia candida (Collembola, Isotomidae).CrossRef | 1:CAS:528:DC%2BD2MXit1WmsLo%3D&md5=d31c9b1f81a12ce4ccafc4232fdd79adCAS | open url image1

Hajibabaei, M., DeWaard, J. R., Ivanova, N. V., Ratnasingham, S., Dooh, R. T., Kirk, S. L., Mackie, P. M., and Hebert, P. D. N. (2005). Critical factors for assembling a high volume of DNA barcodes. Philosophical Transactions of the Royal Society B-Biological Sciences 360, 1959–1967.
Critical factors for assembling a high volume of DNA barcodes.CrossRef | 1:CAS:528:DC%2BD2MXhtlSjsrbE&md5=c3225e33fb170df0244e088aa8791414CAS | open url image1

Hajibabaei, M., Janzen, D. H., Burns, J. M., Hallwachs, W., and Hebert, P. D. N. (2006). DNA barcodes distinguish species of tropical Lepidoptera. Proceedings of the National Academy of Sciences of the United States of America 103, 968–971.
DNA barcodes distinguish species of tropical Lepidoptera.CrossRef | open url image1

Hall, T. A. (1999). BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41, 95–98.
| 1:CAS:528:DC%2BD3cXhtVyjs7Y%3D&md5=08d747d827542961412adcc224832fc5CAS | open url image1

Hammond, P. M. (1992). Species inventory. In ‘Global Biodiversity: Status of the Earth’s Living Resources. A Report Compiled by the World Conservation Monitoring Centre’. (Ed. B. Groombridge.) pp. 17–39. (Chapman & Hall: London.)

Hausmann, A., Hebert, P. D. N., Mitchell, A., Rougerie, R., Sommerer, M., Edwards, T., and Young, C. J. (2009). Revision of the Australian Oenochroma vinaria Guenee, 1858 species-complex (Lepidoptera: Geometridae, Oenochrominae): DNA barcoding reveals cryptic diversity and assesses status of type specimen without dissection. Zootaxa 2239, 1–21. open url image1

Hebert, P. D. N., Cywinska, A., Ball, S. L., and DeWaard, J. R. (2003). Biological identifications through DNA barcodes. Proceedings of the Royal Society of London. Series B. Biological Sciences 270, 313–321.
Biological identifications through DNA barcodes.CrossRef | 1:CAS:528:DC%2BD3sXktVWiu7g%3D&md5=bacde07336013937477a1e8839a85156CAS | open url image1

Hogg, I. D., and Hebert, P. D. N. (2004). Biological identification of springtails (Hexapoda: Collembola) from the Canadian Arctic, using mitochondrial DNA barcodes. Canadian Journal of Zoology-Revue Canadienne De Zoologie 82, 749–754.
Biological identification of springtails (Hexapoda: Collembola) from the Canadian Arctic, using mitochondrial DNA barcodes.CrossRef | open url image1

Hopkin, S. P. (1997). ‘Biology of the Springtails (Insecta: Collembola).’ (Oxford University Press: New York & Tokyo.)

Ivanova, N. V., Dewaard, J. R., and Hebert, P. D. N. (2006). An inexpensive, automation-friendly protocol for recovering high-quality DNA. Molecular Ecology Notes 6, 998–1002.
An inexpensive, automation-friendly protocol for recovering high-quality DNA.CrossRef | 1:CAS:528:DC%2BD2sXmsl2ksw%3D%3D&md5=61bcb820336b724ae357909d1bde24b1CAS | open url image1

James, S. W., Porco, D., Decaëns, T., Richard, B., Rougerie, R., and Erséus, C. (2010). DNA barcoding reveals cryptic diversity in Lumbricus terrestris L., 1758 (Clitellata): resurrection of L. herculeus (Savigny, 1826). PLoS ONE 5, e15629.
DNA barcoding reveals cryptic diversity in Lumbricus terrestris L., 1758 (Clitellata): resurrection of L. herculeus (Savigny, 1826).CrossRef | 1:CAS:528:DC%2BC3MXktlSkug%3D%3D&md5=f48d743a697ac6063482ef591aff579bCAS | open url image1

Jordana, R., Schulz, H.-J., and Baquero, E. (2011a). New species of Entomobrya from Germany (Collembola, Entomobryini). Soil Organisms 83, 249–264. open url image1

Jordana, R., Giuga, L., and Baquero, E. (2011b). New species of Entomobrya from Etna mountain, Sicily (Collembola Entomobryomorpha). Redia-Giornale Di Zoologia 94, 35–38. open url image1

Kimura, M. (1980). A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide-sequences. Journal of Molecular Evolution 16, 111–120.
A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide-sequences.CrossRef | 1:CAS:528:DyaL3MXmtFSktg%3D%3D&md5=78555c3dc7e755aed13a5a2a8c5b8747CAS | open url image1

Kovac, L., and Papac, V. (2010). Revision of the genus Neelus Folsom, 1896 (Collembola, Neelida) with the description of two new troglobiotic species from Europe. Zootaxa 2663, 36–52. open url image1

Letunic, I., and Bork, P. (2007). Interactive Tree Of Life (iTOL): an online tool for phylogenetic tree display and annotation. Bioinformatics (Oxford, England) 23, 127–128.
Interactive Tree Of Life (iTOL): an online tool for phylogenetic tree display and annotation.CrossRef | 1:CAS:528:DC%2BD28XhtlGktLzN&md5=c346545c8b00fd40eafd9ef2fafb3e2fCAS | open url image1

Lukic, M., Houssin, C., and Deharveng, L. (2010). A new relictual and highly troglomorphic species of Tomoceridae (Collembola) from a deep Croatian cave. ZooKeys 69, 1–16. open url image1

Lupetti, P., Marsili, L., Focardi, S., and Dallai, R. (1994). Organochlorine compounds in litter-dwelling arthropods: Collembola (Insecta Apterygota) from central Italy. Acta Zoologica Fennica 195, 94–97. open url image1

McGaughran, A., Torricelli, G., Carapelli, A., Frati, F., Stevens, M. I., Convey, P., and Hogg, I. D. (2010). Contrasting phylogeographical patterns for springtails reflect different evolutionary histories between the Antarctic Peninsula and continental Antarctica. Journal of Biogeography 37, 103–119.
Contrasting phylogeographical patterns for springtails reflect different evolutionary histories between the Antarctic Peninsula and continental Antarctica.CrossRef | open url image1

Palacios-Vargas, J. G., and Arbea, J. I. (2009). Neotropical species of Proisotoma (Collembola: Isotomidae), with description of two new cave species from America. Revista Mexicana De Biodiversidad 80, 445–453. open url image1

Palacios-Vargas, J. G., Deharveng, L., and D’Haese, C. A. (2011). The genus Pronura (Collembola: Neanuridae) in South America, with descriptions of two new species and a barcode sequence for one of them. Revue Suisse de Zoologie 118, 197–205. open url image1

Porco, D., Bedos, A., and Deharveng, L. (2010a). Description and DNA barcoding assessment of the new species Deutonura gibbosa (Collembola: Neanuridae: Neanurinae), a common springtail of Alps and Jura. Zootaxa 2639, 59–68. open url image1

Porco, D., Rougerie, R., Deharveng, L., and Hebert, P. (2010b). Coupling non-destructive DNA extraction and voucher retrieval for small soft-bodied arthropods in a high-throughput context: the example of Collembola. Molecular Ecology Resources 10, 942–945.
Coupling non-destructive DNA extraction and voucher retrieval for small soft-bodied arthropods in a high-throughput context: the example of Collembola.CrossRef | 1:CAS:528:DC%2BC3cXhsVGmtrjP&md5=b3ebfab201623305abac19a2e865b6d7CAS | open url image1

Porco, D., Potapov, M., Bedos, A., Busmachiu, G., Weiner, W. M., Hamra-Kroua, S., and Deharveng, L. (2012a). Cryptic diversity in the ubiquist species Parisotoma notabilis (Collembola, Isotomidae): a long used chimeric species? PLoS ONE 7, e46056.
| 1:CAS:528:DC%2BC38XhsVOlsr%2FP&md5=3c6627d89a3442e93834cef9df257126CAS | open url image1

Porco, D., Decaëns, T., Deharveng, L., James, S., Skarżyński, D., Erséus, C., Butt, K., Richard, B., and Hebert, P. N. (2012b). Biological invasions in soil: DNA barcoding as a monitoring tool in a multiple taxa survey targeting European earthworms and springtails in North America. Biological Invasions , .
Biological invasions in soil: DNA barcoding as a monitoring tool in a multiple taxa survey targeting European earthworms and springtails in North America.CrossRef | open url image1

Potapov, M. (2001). Synopses on Palaearctic Collembola. Volume 3. Isotomidae. Abhandlungen und Berichte des Naturkundemuseums Goerlitz 73, 1–603. open url image1

Riepert, F., and Kula, C. (1996). Development of laboratory methods for testing effects of chemicals and pesticides on Collembola and earthworms. In ‘Communications from the Federal Biological Institute for Agriculture and Forestry’. (Ed. Berlin-Dahlem.) pp. 82.

Rocha-Ramírez, A., Ramirez-Rojas, A., Chavez-Lopez, R., and Alcocer, J. (2007). Invertebrate assemblages associated with root masses of Eichhornia crassipes (Mart.) Solms-Laubach 1883 in the Alvarado Lagoonal System, Veracruz, Mexico. Aquatic Ecology 41, 319–333.
Invertebrate assemblages associated with root masses of Eichhornia crassipes (Mart.) Solms-Laubach 1883 in the Alvarado Lagoonal System, Veracruz, Mexico.CrossRef | open url image1

Rost, M. M. (2006). Ultrastructural changes in the midgut epithelium in Podura aquatica L. (Insecta, Collembola, Arthropleona) during regeneration. Arthropod Structure & Development 35, 69–76.
Ultrastructural changes in the midgut epithelium in Podura aquatica L. (Insecta, Collembola, Arthropleona) during regeneration.CrossRef | open url image1

Rougerie, R., Decaens, T., Deharveng, L., Porco, D., James, S. W., Chang, C. H., Richard, B., Potapov, M., Suhardjono, Y., and Hebert, P. D. N. (2009). DNA barcodes for soil animal taxonomy. Pesquisa Agropecuaria Brasileira 44, 789–802.
DNA barcodes for soil animal taxonomy.CrossRef | open url image1

Saitou, N., and Nei, M. (1987). The neighbor-joining method – a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution 4, 406–425.
| 1:STN:280:DyaL1c7ovFSjsA%3D%3D&md5=9007f6a38c575e135323ec5b129ae4d3CAS | open url image1

Salmon, S. (2001). Earthworm excreta (mucus and urine) affect the distribution of springtails in forest soils. Biology and Fertility of Soils 34, 304–310.
Earthworm excreta (mucus and urine) affect the distribution of springtails in forest soils.CrossRef | 1:CAS:528:DC%2BD3MXnvFOltb8%3D&md5=fc0deaa2cf5bf5d34fd59996db7f15fbCAS | open url image1

Soto-Adames, F. N. (2002). Molecular phylogeny of the Puerto Rican Lepidocyrtus and Pseudosinella (Hexapoda: Collembola), a validation of Yoshii’s “color pattern species”. Molecular Phylogenetics and Evolution 25, 27–42.
Molecular phylogeny of the Puerto Rican Lepidocyrtus and Pseudosinella (Hexapoda: Collembola), a validation of Yoshii’s “color pattern species”.CrossRef | 1:CAS:528:DC%2BD38XnsVeqs74%3D&md5=2ebb78778d61a3aa7d0a8c947ccb39f9CAS | open url image1

Stevens, M. I., and Hogg, I. D. (2006). Contrasting levels of mitochondrial DNA variability between mites (Penthalodidae) and springtails (Hypogastruridae) from the Trans-Antarctic Mountains suggest long-term effects of glaciation and life history on substitution rates, and speciation processes. Soil Biology & Biochemistry 38, 3171–3180.
Contrasting levels of mitochondrial DNA variability between mites (Penthalodidae) and springtails (Hypogastruridae) from the Trans-Antarctic Mountains suggest long-term effects of glaciation and life history on substitution rates, and speciation processes.CrossRef | 1:CAS:528:DC%2BD28XpvFens7s%3D&md5=d67678824613ee96eec1295dc239a198CAS | open url image1

Stevens, M. I., Greenslade, P., Hogg, I. D., and Sunnucks, P. (2006). Southern Hemisphere springtails: could any have survived glaciation of Antarctica? Molecular Biology and Evolution 23, 874–882.
Southern Hemisphere springtails: could any have survived glaciation of Antarctica?CrossRef | 1:CAS:528:DC%2BD28XkvVeqtLg%3D&md5=e8d7cae2d8f2032d6b654c6f12c88ee5CAS | open url image1

Stevens, M. I., Porco, D., D’Haese, C. A., and Deharveng, L. (2011). Comment on “Taxonomy and the DNA Barcoding Enterprise” by Ebach (2011). Zootaxa 2839, 85–88. open url image1

Tamura, K., Dudley, J., Nei, M., and Kumar, S. (2007). MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biology and Evolution 24, 1596–1599.
MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0.CrossRef | 1:CAS:528:DC%2BD2sXpsVGrsL8%3D&md5=d2738b052fcd210827e9d4fc638cc7d5CAS | open url image1

Torricelli, G., Carapelli, A., Convey, P., Nardi, F., Boore, J. L., and Frati, F. (2010). High divergence across the whole mitochondrial genome in the “pan-Antarctic” springtail Friesea grisea: evidence for cryptic species? Gene (Amsterdam) 449, 30–40.
High divergence across the whole mitochondrial genome in the “pan-Antarctic” springtail Friesea grisea: evidence for cryptic species?CrossRef | 1:CAS:528:DC%2BD1MXhtleku7jK&md5=fae2222c7dd1daf8329ba6eabeb17dfbCAS | open url image1

Tully, T., D’Haese, C. A., Richard, M., and Ferriere, R. (2006). Two major evolutionary lineages revealed by molecular phylogeny in the parthenogenetic collembola species Folsomia candida. Pedobiologia 50, 95–104.
Two major evolutionary lineages revealed by molecular phylogeny in the parthenogenetic collembola species Folsomia candida.CrossRef | 1:CAS:528:DC%2BD28XntlWhtLk%3D&md5=fdff06273fbacc874ca8d9d8c81a3d70CAS | open url image1



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