Hostname: page-component-5d59c44645-mrcq8 Total loading time: 0 Render date: 2024-03-01T23:29:29.032Z Has data issue: false hasContentIssue false

Novel microsatellite DNA markers indicate strict parthenogenesis and few genotypes in the invasive willow sawfly Nematus oligospilus

Published online by Cambridge University Press:  29 August 2012

V. Caron*
Australian Centre for Biodiversity, School of Biological Sciences, Monash University, Clayton, Victoria 3800, Australia
M. Norgate
Australian Centre for Biodiversity, School of Biological Sciences, Monash University, Clayton, Victoria 3800, Australia
F.J. Ede
Biosciences Research Division,Department of Primary Industries, PO Box 48, Frankston, Victoria 3199, Australia
T. Nyman
Department of Biology, University of Eastern Finland, PO Box 111, FI-80101, Finland Institute of Systematic Botany, University of Zurich, Zollikerstrasse 107, CH-8008 Zurich, Switzerland
P. Sunnucks
Australian Centre for Biodiversity, School of Biological Sciences, Monash University, Clayton, Victoria 3800, Australia
*Author for correspondence Fax: +61 3 9905 5613 E-mail:


Invasive organisms can have major impacts on the environment. Some invasive organisms are parthenogenetic in their invasive range and, therefore, exist as a number of asexual lineages (=clones). Determining the reproductive mode of invasive species has important implications for understanding the evolutionary genetics of such species, more especially, for management-relevant traits. The willow sawfly Nematus oligospilus Förster (Hymenoptera: Tenthredinidae) has been introduced unintentionally into several countries in the Southern Hemisphere where it has subsequently become invasive. To assess the population expansion, reproductive mode and host-plant relationships of this insect, microsatellite markers were developed and applied to natural populations sampled from the native and expanded range, along with sequencing of the cytochrome-oxidase I mitochondrial DNA (mtDNA) region. Other tenthredinids across a spectrum of taxonomic similarity to N. oligospilus and having a range of life strategies were also tested. Strict parthenogenesis was apparent within invasive N. oligospilus populations throughout the Southern Hemisphere, which comprised only a small number of genotypes. Sequences of mtDNA were identical for all individuals tested in the invasive range. The microsatellite markers were used successfully in several sawfly species, especially Nematus spp. and other genera of the Nematini tribe, with the degree of success inversely related to genetic divergence as estimated from COI sequences. The confirmation of parthenogenetic reproduction in N. oligospilus and the fact that it has a very limited pool of genotypes have important implications for understanding and managing this species and its biology, including in terms of phenotypic diversity, host relationships, implications for spread and future adaptive change. It would appear to be an excellent model study system for understanding evolution of invasive parthenogens that diverge without sexual reproduction and genetic recombination.

Research Paper
Copyright © Cambridge University Press 2012

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)


Atkinson, I.a.E. & Cameron, E.K. (1993) Human influence on the terrestrial biota and biotic communities of New Zealand. Trends in Ecology and Evolution 8, 447451.Google Scholar
Barbarà, T., Palma-Silva, C., Paggi, G.M., Bered, F., Fay, M.F. & Lexer, C. (2007) Cross-species transfer of nuclear microsatellite markers: potential and limitations. Molecular Ecology 16, 37593767.Google Scholar
Benson, R.B. (1950) An introduction to the natural history of British sawflies (Hymenoptera: Symphyta). Transactions of the Society for British Entomology 10, 45134.Google Scholar
Berry, J.A. (1997) Nematus oligospilus (Hymenoptera:Tenthredinidae), a recently introduced sawfly defoliating willows in New Zealand. New Zealand Entomologist 20, 5154.Google Scholar
Boévé, J.L. (2004) Sawflies (Hymenoptera: Tenthredinidae). pp. 1949–953 in Capinera, J.L. (Ed.) Encyclopedia of Entomology. Dordrecht, The Netherlands, Kluwer Academic Publishers.Google Scholar
Bruzzese, E. & McFadyen, R. (2006) Arrival of the leaf-feeding willow sawfly Nematus oligospilus Förster in Australia – pest or beneficial. Plant Protection Quarterly 21, 4344.Google Scholar
Budde, K.B., Gallo, L., Marchelli, P., Mosner, E., Liepelt, S., Ziegenhagen, B. & Leyer, I. (2011) Wide spread invasion without sexual reproduction? A case study on European willows in Patagonia, Argentina. Biological Invasions 13, 4554.Google Scholar
Carl, K.P. (1972) On the biology, ecology and population dynamics of Caliroa cerasi (L.) (Hym., Tenthredinidae). Zeitschrift für Angewandte Entomologie 71, 5883.Google Scholar
Caron, V. (2011) Ecology and evolution of the invasive willow sawfly Nematus oligospilus Förster. School of Biological Sciences, Clayton, Victoria, Australia, Monash University.Google Scholar
Carr, T.G., Roininen, H. & Price, P.W. (1998) Oviposition preference and larval performance of Nematus oligospilus (Hymenoptera: Tenthredinidae) in relation to host plant vigor. Environmental Entomology 27, 615625.Google Scholar
Chambers, G.K. & MacAvoy, E.S. (2000) Microsatelllites: consensus and controversy. Comparative Biochemistry and Physiology Part B 126, 455476.Google Scholar
Charles, J.G. & Allan, D.J. (2000) Development of the willow sawfly, Nematus oligospilus, at different temperatures, and an estimation of voltinism throughout New Zealand. New Zealand Journal of Zoology 27, 197200.Google Scholar
Charles, J.G., Allan, D.J. & Fung, L. (1998) Susceptibility of willows to oviposition by the willow sawfly, Nematus oligospilus. pp. 230234 in Proceedings of the 51st New Zealand Plant Protection Conference. 11–13 August 1998, Hamilton, New Zealand.Google Scholar
Corrie, A.M., Crozier, R.H., Van Heeswijck, R. & Hoffmann, A.A. (2002) Clonal reproduction and population genetic structure of grape phylloxera, Daktulosphaira vitifoliae, in Australia. Heredity 88, 203211.Google Scholar
Courchamp, F., Chapuis, J.L. & Pascal, M. (2003) Mammal invaders on islands: impact, control and control impact. Biological Reviews 78, 347383.Google Scholar
Cowie, B. (2006) Overcoming the threat posed by willow sawfly Nematus oligospilus. A review of research needs and possible options. Christchurch, New Zealand, Environmental Management Services Ltd.Google Scholar
Craig, T.P. & Mopper, S. (1993) Sex ratio variation in sawflies. pp. 6193in Wagner, M.R. & Raffa, K.F. (Eds) Sawfly Life History Adaptations to Woody Plants. San Diego, CA, USA, Academic Press.Google Scholar
Craig, T.P., Price, P.W. & Itami, J.K. (1992) Facultative sex-ratio shifts by a herbivorous insect in response to variation in host plant quality. Oecologia 92, 153161.Google Scholar
Crawford, K.M. & Whitney, K.D. (2010) Population genetic diversity influences colonization success. Molecular Ecology 19, 12531263.Google Scholar
Dapoto, G. & Giganti, H. (1994) Bioecologia de Nematus desantisi Smith (Hymenoptera: Tenthredinidae: Nematinae) en las provincias de Rio Negro y Neuquen (Argentina). Bosque 15, 2732.Google Scholar
Diamond, J.M. (1989) The present, past and future of human-caused extinctions. Philosophical Transactions of the Royal Society of London, Series B 325, 469477.Google Scholar
Dybdahl, M.F. & Lively, C.M. (1995) Diverse, endemic and polyphyletic clones in mixed populations of a freshwater snail (Potamopyrgus antipodarum). Journal of Evolutionary Biology 8, 385398.Google Scholar
Ede, F., Hunt, T., Clements, D. & Caron, V. (2009) Willow sawfly activity in Victoria: 2007–2009. Frankston, Victoria, Australia, Victorian Department of Primary Industries.Google Scholar
Ede, F.J. (2009) Can international experience help us to predict the potential impacts of willow sawfly (Nematus oligospilus Förster) on willow populations in Australia? Plant Protection Quarterly 24, 6266.Google Scholar
Ede, F.J., Caron, V. & Clements, D. (2007) Willow sawfly activity in Victoria: the 2006/07 season. Frankston, Victoria, Australia, Victorian Department of Primary Industries.Google Scholar
Folmer, O., Black, M., Hoeh, W., Lutz, R. & Vrijenhoek, R. (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology 3, 294297.Google Scholar
Gómez, A. & Carvalho, G.R. (2000) Sex, parthenogenesis and genetic structure of rotifers: microsatellite analysis of contemporary and resting egg bank populations. Molecular ecology 9, 203214.Google Scholar
Grosberg, R.K., Levitan, D.R. & Cameron, B.B. (1996) Evolutionary genetics of allorecognition in the colonial hydroid Hyractinia symbiologicarpus. Evolution 50, 22212240.Google Scholar
Hebert, P.D.N., Cywinska, A., Ball, S.L. & Dewaard, J.R. (2003) Biological identifications through DNA barcodes. Proceedings of the Royal Society of London, Series B: Biological Sciences 270, 313321.Google Scholar
Hebert, P.D.N., Penton, E.H., Burns, J.M., Janzen, D.H. & Hallwachs, W. (2004) Ten species in one: DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator. Proceedings of the National Academy of Sciences of the United States of America 101, 1481214817.Google Scholar
Hjalten, J. & Price, P.W. (1997) Can plants gain protection from herbivory by association with unpalatable neighbours?: A field experiment in a willow-sawfly system. Oikos 78, 317322.Google Scholar
Hobbs, R.J. & Mooney, H.A. (2005) Invasive species in a changing world: the interactions between global change and invasives. pp. 310331in Mooney, H.A., Mack, R.N., Mcneely, J.A., Neville, L.E., Schei, P.J. & Waage, J.K. (Eds) Invasive Alien Species-A new Synthesis. Washington, DC, USA, Island Press.Google Scholar
Hurst, L.D., Hamilton, W.D. & Ladle, R.J. (1992) Covert sex. Trends in Ecology and Evolution 7, 144145.Google Scholar
Jokela, J., Lively, C.M., Dybdahl, M.F. & Fox, J.A. (2003) Genetic variation in sexual and clonal lineages of a freshwater snail. Biological Journal of the Linnean Society 79, 165181.Google Scholar
Kawecki, T.J. & Ebert, D. (2004) Conceptual issues in local adaptation. Ecology Letters 7, 12251241.Google Scholar
Kellner, K. & Heinze, J. (2011) Mechanism of facultative parthenogenesis in the ant Platythyrea punctata. Evolutionary Ecology 25, 7789.Google Scholar
Knerer, G. (1993) Life history diversity in sawflies. pp. 3360in Wagner, M.R. & Raffa, K.F. (Eds) Sawfly Life History Adaptations to Woody Plants. San Diego, CA, USA, Academic Press.Google Scholar
Koch, F. & Smith, D.R. (2000) Nematus oligospilus Förster (Hymenoptera : Tenthredinidae), an introduced willow sawfly in the Southern Hemisphere. Proceedings of the Entomological Society of Washington 102, 292300.Google Scholar
Lambrinos, J.G. (2004) How interactions between ecology and evolution influence contemporary invasion dynamics. Ecology 85, 20612070.CrossRefGoogle Scholar
Lee, C.E. (2002) Evolutionary genetics of invasive species. Trends in Ecology and Evolution 17, 386391.CrossRefGoogle Scholar
Levine, J.M., Vila, M., D'antonio, C.M., Dukes, J.S., Grigulis, K. & Lavorel, S. (2003) Mechanisms underlying the impacts of exotic plant invasions. Proceedings of the Royal Society, Series B: Biological Sciences 270, 775781.Google Scholar
Liston, A.D. (1995) Compendium of European Aawflies. Gottfrieding, Germany, Chalastos Forestry.Google Scholar
Lonsdale, W.M. (1999) Global patterns of plant invasions and the concept of invasibility. Ecology 80, 15221536.Google Scholar
Lowe, A., Harris, S. & Ashton, P. (2004) Ecological Genetics: Design, Analysis and Application. Carlton, Victoria, Australia, Blackwell Publishing.Google Scholar
Loxdale, H.D. (2008) The nature and reality of the aphid clone: genetic variation, adaptation and evolution. Agricultural and Forest Entomology 10, 8190.CrossRefGoogle Scholar
Loxdale, H.D. (2009) What's in a clone: the rapid evolution of aphid asexual lineages in relation to geography, host plant adaptation and resistance to pesticides. pp. 535557in Schön, I., Martens, K. & Van Dijk, P.J. (Eds) Lost Sex: The Evolutionary Biology of Parthenogenesis. Berlin, Germany, Springer-Verlag.Google Scholar
Loxdale, H.D. (2010) Rapid genetic changes in natural insect populations. Ecological Entomology 35, 155164.Google Scholar
Lucek, K., Roy, D., Bezault, E., Sivasundar, A. & Seehausen, O. (2010) Hybridization between distant lineages increases adaptive variation during a biological invasion: stickleback in Switzerland. Molecular Ecology 19, 39954011.Google Scholar
Lynch, M. (1985) Spontaneous mutations for life-history characters in an obligate parthenogen. Evolution 39, 804818.Google Scholar
Mack, R.N., Simberloff, D., Lonsdale, W.M., Evans, H., Clout, M. & Bazzaz, F.A. (2000) Biotic invasions: causes, epidemiology, global consequences, and control. Ecological Applications 10, 689710.Google Scholar
MacQuarrie, C.J.K., Langor, D.W. & Sperling, F.a.H. (2007) Mitochondrial DNA variation in two invasive birch leaf-mining sawflies in North America. The Canadian Entomologist 139, 545553.Google Scholar
Maynard Smith, J. (1978) The Evolution of Sex. Oxford, UK, Cambridge University Press.Google Scholar
McGeoch, M.A. & Price, P.W. (2005) Scale-dependent mechanisms in the population dynamics of an insect herbivore. Oecologia 144, 278288.Google Scholar
Mergeay, J., Verschuren, D. & De Meester, L. (2006) Invasion of an asexual American water flea clone throughout Africa and rapid displacement of a native sibling species. Proceedings of the Royal Society, Series B: Biological Sciences 273, 28392844.Google Scholar
Moran, N.A. (1992) The evolution of aphid life cycles. Annual Review of Entomology 37, 321348.Google Scholar
Müller, C., Zwaan, B.J., De Vos, H. & Brakefield, P.M. (2003) Chemical defence in a sawfly: genetic components of variation in relevant life-history traits. Heredity 90, 468475.Google Scholar
Müller, C., Barker, A., Boeve, J.L., De Jong, P.W., De Vos, H. & Brakefield, P.M. (2004) Phylogeography of two parthenogenetic sawfly species (Hymenoptera: Tenthredinidae): relationship of population genetic differentiation to host plant distribution. Biological Journal of the Linnean Society 83, 219227.Google Scholar
Naumann, I.D., Williams, M.A. & Schmidt, S. (2002) Synopsis of the Tenthredinidae (Hymenoptera) in Australia, including two newly recorded, introduced sawfly species associated with willows (Salix spp.). Australian Journal of Entomology 41, 16.Google Scholar
Normark, B.B. (2003) The evolution of alternative genetic systems in insects. Annual Review of Entomology 48, 397423.Google Scholar
Nyman, T. (2002) The willow bud galler Euura mucronata Hartig (Hymenoptera: Tenthredinidae): one polyphage or many monophages? Heredity 88, 288295.Google Scholar
Nyman, T., Farrell, B.D., Zinovjev, A.G. & Vikberg, V. (2006a) Larval habits, host-plant associations, and speciation in nematine sawflies (Hymenoptera: Tenthredinidae). Evolution 60, 16221637.Google Scholar
Nyman, T., Zinovjev, A.G., Vikberg, V. & Farrell, B.D. (2006b) Molecular phylogeny of the sawfly subfamily Nematinae (Hymenoptera : Tenthredinidae). Systematic Entomology 31, 569583.Google Scholar
Nyman, T., Vikberg, V., Smith, D.R. & Boevé, J.-L. (2010) How common is ecological speciation in plant-feeding insects? A ‘Higher’ Nematinae perspective. BMC Evolutionary Biology 10, 266.CrossRefGoogle ScholarPubMed
Palma-Silva, C., Cavallari, M.M., Barbará, T., Lexer, C., Gimenes, M.A., Bered, F. & Bodanese-Zanetti, M.H. (2007) A set of polymorphic microsatellite loci for Vriesea gigantea and Alcantarea imperialis (Bromeliaceae) and cross-amplification in other bromeliad species. Molecular Ecology Notes 7, 654657.Google Scholar
Park, D.-S., Foottit, R., Maw, E. & Hebert, P.D.N. (2011) Barcoding Bugs: DNA-Based Identification of the True Bugs (Insecta: Hemiptera: Heteroptera). Plos one 6, e18749.Google Scholar
Peakall, R. & Smouse, P.E. (2006) GENALEX 6: Genetic analysis in Excel. Population genetic software for teaching and research. Molecular Ecology Notes 6, 288295.CrossRefGoogle Scholar
Peccoud, J., Figueroa, C.C., Silva, A.X., Ramirez, C.C., Mieuzet, L., Bonhomme, J., Stoeckel, S., Plantegenest, M. & Simon, J.C. (2008) Host range expansion of an introduced insect pest through multiple colonizations of specialized clones. Molecular Ecology 17, 46084618.Google Scholar
Price, P.W. & Hunter, M.D. (2005) Long-term population dynamics of a sawfly show strong bottom-up effects. Journal of Animal Ecology 74, 917925.CrossRefGoogle Scholar
Price, P.W., Roininen, H. & Ohgushi, T. (2005) Adaptive radiation into ecological niches with eruptive dynamics: a comparison of tenthredinid and diprionid sawflies. Journal of Animal Ecology 74, 397408.Google Scholar
Primmer, C.R., Møller, A.P. & Ellegren, H. (1996) A wide-range survey of cross-species microsatellite amplification in birds. Molecular Ecology 5, 365378.Google Scholar
Primmer, C.R., Painter, J.N., Koskinen, M.T., Palo, J.U. & Merilä, J. (2005) Factors affecting cross-species microsatellite amplification. Journal of Avian Biology 36, 348360.Google Scholar
Pschorn-Walcher, H. (1982) Unterordnung Symphyta, Pflanzenwespen. pp. 4234in Schwenke, W. (Ed.) Die Forstschadlinge Europas, Bd. 4. Hautflugler und Zweiflugler. Hamburg, Germany, P. Parey.Google Scholar
Rabeling, C., Lino-Neto, J., Cappellari, S.C., Dos-Santos, I.A., Mueller, U.G. & Bacci, M. (2009) Thelytokous parthenogenesis in the fungus-gardening ant Mycocepurus smithii (Hymenoptera: Formicidae). Plos one 4, e6781.Google Scholar
Raven, P.H. & Yeates, D.K. (2007) Australian biodiversity: threats for the present, opportunities for the future. Australian Journal of Entomology 46, 177187.Google Scholar
Roberts, J.M.K. & Weeks, A.R. (2009) Permanent genetic resources added to Molecular Ecology Resources Database 1 May 2009–31 July 2009. Molecular Ecology Resources 9, 14601466.Google Scholar
Roininen, H., Price, P.W. & Tahvanainen, J. (1996) Bottom-up and top-down influences in the trophic system of a willow, a galling sawfly, parasitoids and inquilines. Oikos 77, 4450.Google Scholar
Roman, J. & Darling, J.A. (2007) Paradox lost: genetic diversity and the success of aquatic invasions. Trends in Ecology and Evolution 22, 454464.Google Scholar
Rozen, S. & Skaletsky, H.J. (2000) Primer3 on the WWW for general users and for biologist programmers. pp. 365386in Krawetz, S. & Misener, S. (Eds) Bioinformatics Methods and Protocols: Methods in Molecular Biology. Totowa, NJ, USA, Humana Press.Google Scholar
Sanders, N.J., Gotelli, N.J., Heller, N.E. & Gordon, D.M. (2003) Community disassembly by an invasive species. Proceedings of the National Academy of Sciences of the United States of America 100, 24742477.Google Scholar
Schlotterer, C. & Tautz, D. (1992) Slippage synthesis of simple sequence DNA. Nucleic Acid Research 20, 211215.CrossRefGoogle ScholarPubMed
Schmidt, S. & Smith, D.R. (2009) Selandriinae, a subfamily of Tenthredinidae new to Australia, and a review of other Australian Tenthredinidae (Hymenoptera: Symphyta). Australian Journal of Entomology 48, 305309.Google Scholar
Selkoe, K.A. & Toonen, R.J. (2006) Microsatellites for ecologists: a practical guide to using and evaluating microsatellite markers. Ecology Letters 9, 615629.Google Scholar
Smith, D.R. (1979) Suborder Symphyta. pp. 3137in Krombein, K.V., Hurd, P.D.J., Smith, D.R. & Burks, B.D. (Eds) Catalog of Hymenoptera in America North of Mexico. Washington, DC, USA, Smithsonian Institution Press.Google Scholar
Sunnucks, P. (2000) Efficient genetic markers for population biology. Trends in Ecology and Evolution 15, 199203.Google Scholar
Sunnucks, P. & Hales, D.F. (1996) Numerous transposed sequences of mitochondrial cytochrome oxidase I–II in aphids of the genus Sitobion (Hemiptera: Aphididae). Molecular Biology and Evolution 13, 510524.Google Scholar
Sunnucks, P., England, P.R., Taylor, A.C. & Hales, D.F. (1996) Microsatellite and chromosome evolution of parthenogenetic Sitobion aphids in Australia. Genetics 144, 747756.Google Scholar
Sunnucks, P., Chisholm, D., Turak, E. & Hales, D.F. (1998) Evolution of an ecological trait in parthenogenetic Sitobion aphids. Heredity 81, 638647.Google Scholar
Suomalainen, E. (1962) Significance of parthenogenesis in the evolution of insects. Annual Review of Entomology 7, 349366.Google Scholar
Tagg, N., Innes, D.J. & Doncaster, C.P. (2005) Outcomes of reciprocal invasions between genetically diverse and genetically uniform populations of Daphnia obtusa (Kurz). Oecologia 143, 527536.Google Scholar
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. (2011) MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution 28, 27312739.Google Scholar
Urban, A.J. & Eardley, C.D. (1995) A recently introduced sawfly, Nematus oligospilus Forster (Hymenoptera: Tenthredinidae), that defoliates willows in southern Africa. African Entomology 3, 2327.Google Scholar
Vorburger, C., Lancaster, M. & Sunnucks, P. (2003) Environmentally related patterns of reproductive modes in the aphid Myzus persicae and the predominance of two ‘superclones’ in Victoria, Australia. Molecular Ecology 12, 34933504.Google Scholar
Wilson, A.C.C., Sunnucks, P., Blackman, R.L. & Hales, D.F. (2002) Microsatellite variation in cyclically parthenogenetic populations of Myzus persicae in south-eastern Australia. Heredity 88, 258266.Google Scholar
Wilson, A.C.C., Sunnucks, P. & Hales, D.F. (2003) Heritable genetic variation and potential for adaptive evolution in asexual aphids (Aphidoidea). Biological Journal of the Linnean Society 79, 115135.Google Scholar
Wilson, A.C.C., Massonnet, B., Simon, J.C., Prunier-Leterme, N., Dolatti, L., Llewellyn, K.S., Figueroa, C.C., Ramirez, C.C., Blackman, R.L., Estoup, A. & Sunnucks, P. (2004) Cross-species amplification of microsatellite loci in aphids: assessment and application. Molecular Ecology Notes 4, 104109.Google Scholar
Zane, L., Bargelloni, L. & Patarnello, T. (2002) Strategies for microsatellite isolation: a review. Molecular Ecology 11, 116.Google Scholar
Zenger, K.R., Eldridge, M.D.B., Pope, L.C. & Cooper, D.W. (2003) Characterisation and cross-species utility of microsatellite markers within kangaroos, wallabies and rat kangaroos (Macropodoidea: Marsupialia). Australian Journal of Zoology 51, 587596.Google Scholar