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Mitochondrial DNA suggests cryptic speciation in Prodiplosis longifila Gagné (Diptera: Cecidomyiidae) associated with geographic distance and host specialization

  • D.N. Duque-Gamboa (a1), M.F. Castillo-Cárdenas (a1), L.M. Hernández (a2), Y.C. Guzmán (a2), M.R. Manzano (a2) and N. Toro-Perea (a1)...


Prodiplosis longifila is reported as a pest of a wide range of species cultivated in America, including citrus, solanaceous species and asparagus. This species has different behavioural traits that are primarily centred on the oviposition habit and the feeding of larvae, which can change depending on the host. However, scarce information is available on population studies and the natural history of this insect, and uncertainty exists about the taxonomic identity and the geographic distribution of this species. The main objective was to perform a phylogenetic and genetic study of P. longifila populations and to define whether the North American and South American populations belong to the same species or whether a differentiation process had occurred due to geographic distance. A second objective was to determine whether this species showed genetic differentiation by host specialization in South America. The phylogenetic and population analyses based on DNA barcodes (cytochrome oxidase I gene) and a region of the ribosomal DNA (ITS2) revealed divergent clades attributable to geographic distance and host specificity. The North American and South American P. longifila insects were confirmed to be genetically distinct, and the genetic distances exceeded the values expected for intraspecific variation. In South America, the population analysis of P. longifila from tomato, sweet pepper (Solanaceae), Tahiti lime and key lime (Rutaceae) hosts evidenced high genetic differentiation between populations associated with different hosts and an absence of gene flow between these groups, suggesting the corresponding formation of cryptic species.


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Aide, T.M., Clark, M.L., Grau, H.R., López-Carr, D., Levy, M.A., Redo, D., Bonilla-Moheno, M., Riner, G., Andrade-Núñez, M.J. & Muñiz, M. (2013) Deforestation and reforestation of Latin America and the Caribbean (2001–2010). Biotropica 45, 262271.
Berlocher, S.H. & Feder, J.L. (2002) Sympatric speciation in phytophagous insects: moving beyond controversy? Annual Review of Entomology 47, 773815.
Bickford, D., Lohman, D.J., Sodhi, N.S., Ng, P.K.L., Meier, R., Winker, K., Ingram, K.K. & Das, I. (2007) Cryptic species as a window on diversity and conservation. Trends in Ecology & Evolution 22, 148155.
Bolnick, D.I. & Fitzpatrick, B.M. (2007) Sympatric speciation: models and empirical evidence. Annual Review of Ecology, Evolution, and Systematics 38, 459487.
Bush, G.L. (1975) Modes of animal speciation. Annual Review of Ecology and Systematics 6, 339364.
Collins, F.H. & Paskewitz, S.M. (1996) A review of the use of ribosomal DNA (rDNA) to differentiate among cryptic Anopheles species. Insect Molecular Biology 5, 19.
Cook, M.A., Ozeroff, S.N., Fitzpatrick, S.M. & Roitberg, B.D. (2011) Host-associated differentiation in reproductive behaviour of cecidomyiid midges on cranberry and blueberry. Entomologia Experimentalis et Applicata 141, 814.
Darriba, D., Taboada, G.L., Doallo, R. & Posada, D. (2012) Jmodeltest 2: more models, new heuristics and parallel computing. Nature Methods 9, 772772.
Dent, D. (2000) Insect Pest Management. Wallingford, UK, CABI Publishing.
Dorchin, N., Scott, E.R., Clarkin, C.E., Luongo, M.P., Jordan, S. & Abrahamson, W.G. (2009) Behavioural, ecological and genetic evidence confirm the occurrence of host-associated differentiation in goldenrod gall-midges. Journal of Evolutionary Biology 22, 729739.
Drès, M. & Mallet, J. (2002) Host races in plant–feeding insects and their importance in sympatric speciation. Philosophical Transactions of the Royal Society of London B: Biological Sciences 357, 471492.
Edgar, R.C. (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research 32, 17921797.
Excoffier, L., Laval, G. & Schneider, S. (2005) Arlequin (version 3.0): an integrated software package for population genetics data analysis. Evolutionary Bioinformatics Online 1, 4750.
Folmer, O., Black, M., Hoeh, W. & Lutz, R. (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology 3, 294299.
Gagné, R.J. (1986 a) Revision of Prodiplosis (Diptera: Cecidomyiidae) with descriptions of three new species. Annals of the Entomological Society of America 79, 235245.
Gagné, R.J. (1986 b) A new Nearctic species of Contarinia (Diptera: Cecidomyiidae) recently introduced into Hawaii on Cupressus (Cupressaceae). Proceedings of the Entomological Society of Washington 88, 127130.
Gagné, R.J. (1994) The Gall Midges of the Neotropical Region. Ithaca, New York, Cornell University Press.
Gagné, R.J. & Jaschhof, M. (2014) A Catalog of the Cecidomyiidae (Diptera) of the World. 3rd edn. Digital version 2. Washington, USA, USDA.
Guillemaud, T., Ciosi, M., Lombaert, E. & Estoup, A. (2011) Biological invasions in agricultural settings: insights from evolutionary biology and population genetics. Comptes Rendus Biologies 334, 237246.
Guindon, S. & Gascuel, O. (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Systematic Biology 52, 696704.
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, 9598.
Hebert, P.D.N., Cywinska, A. & Ball, S.L. (2003) Biological identifications through DNA barcodes. Proceedings of the Royal Society of London B: Biological Sciences 270, 313321.
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.
Hernandez, L.M., Guzman, Y.C., Martínez-Arias, A., Manzano, M.R. & Selvaraj, J.J. (2015) The bud midge Prodiplosis longifila: damage characteristics, potential distribution and presence on a new crop host in Colombia. SpringerPlus 4, 205.
Ji, Y., Zhang, D. & He, L. (2003) Evolutionary conservation and versatility of a new set of primers for amplifying the ribosomal internal transcribed spacer regions in insects and other invertebrates. Molecular Ecology Notes 3, 581585.
Jinbo, U., Kato, T. & Ito, M. (2011) Current progress in DNA barcoding and future implications for entomology. Entomological Science 14, 107124.
Kanno, H. & Harris, M.O. (2000) Both chemical and physical features of grass leaves influence host selection by the Hessian fly. Journal of Chemical Ecology 26, 23352354.
Kikkert, J.R., Hoepting, C.A., Wu, Q., Wang, P., Baur, R. & Shelton, A.M. (2006) Detection of Contarinia nasturtii (Diptera: Cecidomyiidae) in New York, a new pest of cruciferous plants in the United States. Journal of Economic Entomology 99, 13101315.
Lampo, M. & Medialdea, V. (1994) Dispersal pattern of the sorghum midge (Diptera: Cecidomyiidae) in sorghum plots. Environmental Entomology 23, 551555.
Librado, P. & Rozas, J. (2009) DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25, 14511452.
Mathur, S., Cook, M.A., Sinclair, B.J. & Fitzpatrick, S.M. (2012) DNA barcodes suggest cryptic speciation in Dasineura oxycoccana (Diptera: Cecidomyiidae) on cranberry, Vaccinium macrocarpon, and blueberry, V. corymbosum. Florida Entomologist 95, 387394.
Miao, J., Wu, Y.Q., Gong, Z.J., He, Y.Z., Duan, Y. & Jiang, Y.L. (2013) Long-distance wind-borne dispersal of Sitodiplosis mosellana Géhin (Diptera:Cecidomyiidae) in Northern China. Journal of Insect Behavior 26, 120129.
Olson, D.M., Dinerstein, E., Wikramanayake, E.D., Burgess, N.D., Powell, G.V.N., Underwood, E.C., D'amico, J.A., Itoua, I., Strand, H.E. & Morrison, J.C. (2001) Terrestrial ecoregions of the world: a new map of life on earth: a new global map of terrestrial ecoregions provides an innovative tool for conserving biodiversity. BioScience 51, 933938.
Peakall, R.O.D. & Smouse, P.E. (2006) GENALEX 6: genetic analysis in excel. Population genetic software for teaching and research. Molecular Ecology Notes 6, 288295.
Pendleton, B.B. & Teetes, G.L. (1994) Sorghum midge dispersal from sorghum. Southwestern Entomologist 19, 110.
Pérez-Portela, R., Arranz, V., Rius, M. & Turon, X. (2013) Cryptic speciation or global spread? The case of a cosmopolitan marine invertebrate with limited dispersal capabilities. Scientific Reports 3, 3197.
Pizano, C. & Garcia, H. (2014) El bosque seco tropical en Colombia. Bogotá, DC, Colombia, Instituto de Investigación de Recursos Biológicos Alexander von Humboldt.
Posada, D. & Crandall, K.A. (2001) Intraspecific gene genealogies: trees grafting into networks. Trends in Ecology & Evolution 16, 3745.
QGIS Development Team (2017) QGIS Geographic Information System. Open Source Geospatial Foundation Project. Available online at
Richards, T.J., Shanafelt, D.W. & Fenichel, E.P. (2014) Foreclosures and invasive insect spread: the case of Asian citrus psyllid. American Journal of Agricultural Economics 96, 615630.
Stireman, J.O. III, Nason, J.D. & Heard, S.B. (2005) Host-associated genetic differentiation in phytophagous insects: general phenomenon or isolated exceptions? Evidence from a goldenrod-insect community. Evolution 59, 25732587.
Stokes, K., Stiling, P., Gilg, M.R. & Rossi, A.M. (2012) The gall midge Asphondylia borrichiae (Diptera: Cecidomyiidae): an indigenous example of host-associated genetic divergence in sympatry. Environmental Entomology 41, 12461254.
Sylven, E. & Lövgren, L. (1995) Dasineura ingeris sp. n.(Diptera: Cecidomyiidae) on Salix viminalis in Sweden, including comparisons with some other Dasineura species on Salix. Systematic Entomology 20, 5971.
Tamura, K., Stecher, G., Peterson, D., Filipski, A. & Kumar, S. (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution 30, 27252729.
Thomas, Y., Bethenod, M-T., Pelozuelo, L., Frérot, B., Bourguet, D. & Pitnick, S. (2003) Genetic isolation between two sympatric host-plant races of the European corn borer, Ostrinia nubilalis Hübner. I. Sex pheromone, moth emergence timing, and parasitism. Evolution 57, 261273.
Toews, D.P.L. & Brelsford, A. (2012) The biogeography of mitochondrial and nuclear discordance in animals. Molecular Ecology 21, 39073930.
Turelli, M., Barton, N.H. & Coyne, J.A. (2001) Theory and speciation. Trends in Ecology & Evolution 16, 330343.
Valarezo, O., Canarte, E., Navarrete, B. & Arias de Lopez, M. (2003) Prodiplosis longifilia Diptera:(Cecidomyiidae), principal plaga del tomate en el Ecuador. Diagnóstico, Bioecología y Manejo. Iniap, Promsa y Cedege. Manual no. 51. Ecuador.
Via, S. (1999) Reproductive isolation between sympatric races of pea aphids. I. Gene flow restriction and habitat choice. Evolution 53, 14461457.
Via, S. (2001) Sympatric speciation in animals: the ugly duckling grows up. Trends in Ecology & Evolution 16, 381390.
Walker, T.J. (1964) Cryptic species among sound-producing ensiferan Orthoptera (Gryllidae and Tettigoniidae). The Quarterly Review of Biology 39, 345355.
Watterson, G.A. & Guess, H.A. (1977) Is the most frequent allele the oldest? Theoretical Population Biology 11, 141160.
Withers, T.M., Harris, M.O. & Madie, C. (1997) Dispersal of mated female Hessian Flies (Diptera: Cecidomyiidae) in field arrays of host and nonhost plants. Environmental Entomology 26, 12471257.
Yao, H., Song, J., Liu, C., Luo, K., Han, J., Li, Y., Pang, X., Xu, H., Zhu, Y. & Xiao, P. (2010) Use of ITS2 region as the universal DNA barcode for plants and animals. PLoS ONE 5, e13102.
Zhang, D.X. & Hewitt, G.M. (2003) Nuclear DNA analyses in genetic studies of populations: practice, problems and prospects. Molecular Ecology 12, 563584.
Zhang, G.F., Meng, X.Q., Min, L., Qiao, W.N. & Wan, F.H. (2012) Rapid diagnosis of the invasive species, Frankliniella occidentalis (Pergande): a species-specific COI marker. Journal of Applied Entomology 136, 410420.



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