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Type: Article
Published: 2017-11-24
Page range: 467–484
Abstract views: 68
PDF downloaded: 49

Description of two new species closely related to Doryctobracon areolatus (Szépligeti, 1911) (Hymenoptera, Braconidae), based on morphometric and molecular analyses

Departamento de Entomologia e Acarologia, ESALQ, Universidade de São Paulo (USP), Caixa Postal 9, 13418-900, Piracicaba, SP.
Departamento de Entomologia e Acarologia, ESALQ, Universidade de São Paulo (USP), Caixa Postal 9, 13418-900, Piracicaba, SP.
Universidade Federal de São Carlos UFSCAR, Caixa Postal 676, CEP 13565-905, São Carlos, SP
Departamento de Entomologia e Acarologia, ESALQ, Universidade de São Paulo (USP), Caixa Postal 9, 13418-900, Piracicaba, SP.
Hymenoptera geometric morphometry canonical variate analysis ITS2 28S-D2 rDNA molecular taxonomy


This study clarifies the identity of two morphs that appear similar but not identical to the braconid wasp Doryctobracon areolatus (Szépligeti), by analyses of fore wing morphometry and the ITS2 and D2 region of 28S ribosomal DNA. Wing measurements of D. areolatus and the two morphs, one with a yellow stigma and one with a brown stigma, from several Brazilian localities, were taken and subjected to geometric morphometry. Evaluation of 20 anatomical points on the fore wing by means of multivariate analysis revealed that these morphs consistently differ from D. areolatus. Intraspecific size variation in both molecular markers (ITS2 and 28S-D2 rDNA) was also detected among populations of D. areolatus from the states of Amapá, São Paulo, Goiás, and Tocantins, but no such difference was observed among samples of the two morphs. However, high sequence variability was observed for both markers among D. areolatus and these morphs. Morphometric analysis yielded similar results and produced dendrogram congruent with those based on the molecular markers, also indicating that D. areolatus corresponds to a complex of cryptic species. Therefore, our morphometric and molecular data, in addition to revealing that D. areolatus includes cryptic species, showed that the unidentified morphs actually represent two distinct, previously undescribed species of Doryctobracon. These new taxa are formally described herein, as D. whartoni sp. nov. and D. adaimei sp. nov.



  1. Adaime, R., Nascimento, D.B., Deus, E.G., Souza, G.D. & Oliveira, L.P.S. (2007) Hospedeiros e parasitóides de Anastrepha spp. (Diptera: Tephritidae) em Itaubal do Piririm, Estado do Amapá, Brasil. Ciência Rural, 37, 557–560.

    Aljanabi, S.M. & Martinez, I. (1997) Universal and rapid salt-extraction of high quality genomic DNA for PCR-based techniques. Nucleic Acids Research, 25, 4692–4693.

    Baylac, M., Villemant, C. & Simbolotti, G. (2003) Combining geometric morphometrics with pattern recognition for the investigation of species complexes. Biological Journal of the Linnean Society, 80, 89–98.

    Belshaw, R. & Quicke, D.L.J. (1997) A molecular phylogeny of the Aphidiinae (Hymenoptera: Braconidae). Molecular Phylogenetics and Evolution, 7, 281–293.

    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 and Evolution, 22, 148–155.

    Billah, M.K., Kimani-Njogu, S.W., Wharton, R.A., Wooley, J.B. & Masiga, D. (2008) Comparison of five allopatric fruit fly parasitoid population (Psyttalia species) (Hymenoptera: Braconidae) from coffee fields using morphometric and molecular methods. Bulletin of Entomological Research, 98, 63–75.

    Bomfim, D.A., Uchôa-Fernandes, M.A. & Bragança, M.A.L. (2007) Hosts and Parasitoids of Fruit Flies (Diptera: Tephritoidea) in the State of Tocantins, Brazil. Neotropical Entomology, 36, 984–986.

    Braga Filho, J.R., Veloso, W.R.S., Naves, R.V. & Ferreira, G.A. (2001) Entomofauna associada aos frutos do bacupari, Salacia crassifolia (Mart.) Peyr, nos cerrados do Brasil central. Pesquisa Agropecuária Tropical, 31, 47–54.

    Canal Daza, N.A. & Zucchi, R.A. (2000) Parasitóides - Braconidae. In: Malavasi, A. & Zucchi, R.A. Moscas-das-frutas de importância econômica no Brasil. Conhecimento básico e aplicado, Holos-FAPESP Press, Ribeirão Preto, SP, pp. 119–126.

    Ciociola Jr., A.I., Querino, R.B., Zucchi, R.A. & Stouthamer, R. (2001) Molecular tool for identification of closely related species of Trichogramma (Hymenoptera: Trichogrammatidae): T. rojasi Nagajara & Nagarkatti and T. lasallei Pinto. Neotropical Entomology, 30, 575–578.

    Costa Lima, A. (1937) Vespa do gênero Opius, parasitas de larvas de moscas-das-frutas (Hymenoptera: Braconidae). O Campo, 8, 22–32.

    Deus, E.G. & Adaime, R. (2013) Dez anos de pesquisas sobre moscas-das-frutas (Diptera: Tephritidae) no estado do Amapá: avanços obtidos e desafios futuros. Biota Amazônia, 3, 157–168.

    Deus, E.G., Adaime, R., Nascimento, D.B., Marinho, C.F. & Zucchi, R.A. (2010) Hospedeiros e parasitóides de espécies de Anastrepha (Diptera, Tephritidae) em dois municípios do Estado do Amapá. Revista de Agricultura, 84, 194–203.

    Eitam, T.H., Sivinski, J. & Aluja, M. (2003) Use of Host Fruit Chemical Cues for Laboratory Rearing of Doryctobracon areolatus (Hymenoptera: Braconidae), a Parasitoid of Anastrepha spp. (Diptera: Tephritidae) Avi. The Florida Entomologist, 86, 211–216.[0211:UOHFCC]2.0.CO;2

    Fouly, H.M., Wilkinson, H.T. & Chen, W. (1997) Restriction analysis of internal transcribe spacers and the small subunit gene of ribosomal DNA among four Gaeumannomyces species. Mycologia, 89, 590–597.

    Gallego, D. & Galián, J. (2001) The internal transcribed spacers (ITS1 and ITS2) of the rDNA differentiates the bark beetles forest pests Tomicus destruens and T. piniperda. Insect Molecular Biology, 10, 415–420.

    Greene, E. (1989) A diet-induced developmental polymorphism in a caterpillar. Science, 243, 643–646.

    Jesus, C.R., Pereira, J.D.B., Oliveira, M.N., Silva, R.A., Souza Filho, M.F., Costa Neto, S.V., Marinho, C.F. & Zucchi, R.A. (2008) New records of fruit flies (Diptera: Tephritidae), wild hosts and parasitoids (Hymenoptera: Braconidae) in the Brazilian Amazon. Neotropical Entomology, 37, 733–734.

    Jesus, C.R., Adaime, R., Oliveira, M.N., Silva, W.R., Costa Neto, S.V. & Souza Filho, M.F. (2012) Anastrepha (Diptera: Tephritidae) species, their hosts and parasitoids (Hymenoptera: Braconidae) in five municipalities of the state of Amapá, Brazil. The Florida Entomologist, 95, 694–705.

    Kumar, S., Stecher, G. & Tamura, K. (2016) MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33, 1870–1874.

    Lenormand, T. (2002) Gene flow and the limits to natural selection. Trends in Ecology & Evolution, 17, 183–372.

    Leon, J.H., Logarzo, G.A. & Triapitsyn, S.V. (2006) Preliminary evidence from reproductive compatibility studies suggests that Gonatocerus tuberculifemur as a cryptic species complex, or a new species identified: development and utility of molecular diagnostic markers. In: Pierce’s Disease Research Symposium. San Diego, California, pp. 44–47.

    Leonel Junior, F.L., Zucchi, R.A. & Canal Daza, N.A. (1996) Parasitismo de moscas-das-frutas (Diptera: Tephritidae) por Braconidae (Hymenoptera) em duas localidades do Estado de São Paulo. Anais da Sociedade Entomológica do Brasil, 25, 199–206.

    Monteiro, L.R. & Reis, S.F. (1999) Princípios de morfometria geométrica. Holos Editora, Ribeirão Preto, SP, 188 pp.

    Moraes, D.A. (2003) A Morfometria Geométrica e a “Revolução na Morfometria”: localizando mudanças na forma dos organismos URL. Available from:
    metria_geometrica.pdf (accessed 29 March 2012)

    Mound, L.A. (2005) Fighting, flight and fecundity: Behavioural determinants of Thysanoptera structural diversity. In: Ananthakrishnan, T.N. & Whitman, D.W. (Eds.), Insects and Phenotypic Plasticity. Science Publishers Inc., Enfield, NH, pp. 81–105.

    Müller, T., Philippi, N., Dandekar, T., Schultz, J. & Wolf, M. (2007) Distinguishing species. RNA 13, 1469–1472.

    Murillo, F.D., Cabrera-Mireles H., Barrera, J.F., Liedo, P. & Montoya, P. (2015) Doryctobracon areolatus (Hymenoptera, Braconidae) a parasitoid of early developmental stages of Anastrepha obliqua (Diptera, Tephritidae). Journal of Hymenoptera Research, 46, 91–105.

    Nunes, A.M., Nava, D.E., Müller F.A., Gonçalves, R.S. & Garcia, M.S. (2011) Biology and parasitic potential of Doryctobracon areolatus on Anastrepha fraterculus larvae. Pesquisa Agropecuária Brasileira, 46, 669–671.

    Ovruski, S.M. (2003) Nuevos aportes a la taxonomía de las especies de Opiinae (Hymenoptera: Braconidae), parasitoides de Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae) en la provincia de Tucumán, Argentina. Acta Zoológica Lilloana, 47, 15–44.

    Parra, J.R.P., Botelho, P.S.M., Correa-Ferreira, B.S. & Bento, J.M.S. (2002) Controle biológico no Brasil: parasitóides e predadores. Manole Press, São Paulo, SP, 609 pp.

    Pigliucci, M. (2001) Phenotypic plasticity. In: Fox, C.W., Roff, D.A. & Fairbairn, D.J. Evolutionary Ecology: Concepts and Case Studies. Oxford University Press, New York, NY, pp. 58–69.

    Porter, C.H. & Collins, F.H. (1991) Species-diagnostic differences in a ribosomal DNA Internal Transcribe Spacer from the sibling species Anopheles freeborni and Anopheles hermsi (Diptera: Culicidae). The American Journal of Tropical Medicine and Hygiene, 45, 271–279.

    Rohlf, F.J. (2004) tpsDig Version 1.40. Department of Ecology and Evolution, Stony Brook University, Stone Brook, New York.

    Rohlf, F.J. (2007) tpsRewl Version 1.45. Department of Ecology and Evolution, Stony Brook University. Stony Brook, New York.

    Schlichting, C.D. & Pigliucci, M. (1998) Phenotypic Evolution: A Reaction Norm Perspective. Sinauer Associates Press, Sunderland, MA, 387 pp.

    Sharkey, M.J. & Wharton, R.A. (1997) Morphology and Terminology. In: Wharton, R.A., Marsh, P.M. & Shakey, M.J. (Eds.), Manual of the New World Genera of the Family Braconidae (Hymenoptera), Special Publication of the International Society of Hymenopterists. Vol. l. Allen Press, New York, pp. 19–37.

    Silva, I.M.M.S., Honda, J., Van Kan, F., Hu, J., Neto, L., Pintureau, B. & Stouthamer, R. (1999) Molecular differentiation of five Trichogramma species occurring in Portugal. Biological Control, 16, 177–184.

    Silva, W.R. & Adaime, R. (2007) Levantamento de moscas-das-frutas e de seus parasitóides no município de Ferreira Gomes, Estado do Amapá. Ciência Rural 37, 265–268.

    Silva, R.A., R., Lima, A.L., Xavier, S.L.O., Silva, W.R., Marinho, C.F. & Zucchi, R.A. (2011) Anastrepha species (Diptera: Tephritidae), their hosts and parasitoids in southern Amapá State, Brazil. Biota Neotropica, 11, 431–436.

    Stouthamer, R., Jochemsen, P., Platner, G. & Pinto, J.D. (2000) Crossing incompatibility between Trichogramma minutum and T. platneri (Hymenoptera: Trichogrammatidae): Implications for application in biological control. Environmental Entomology, 29, 832–837.

    Stouthamer, R., Rugman-Jones, P., Thu, P.Q., Eskalen, A., Thibault, T., Hulcr, J. Wang, L.-J, Jordal, B.H., Chen, C.-Y., Cooperband, M., Lin, C.-S., Kamata, N., Lu, S.-S., Masuya, H., Mendel, Z., Rabaglia, R., Sanguansub, S., Shinh, H.-H., Sittichaya, W. & Zong, S. (2017) Tracing the origin of a cryptic invader: phylogeography of the Euwallacea fornicates (Coleoptera: Curculionidae: Scolytinae) species complex. Agricultural and Forest Entomology.

    Slatkin, M. (1973) Gene flow and selection in a cline. Genetics, 75, 733–756.

    Szépligeti, G. (1911) Zwei neue Braconiden aus Brasilien. Bollettino del Laboratorio di Zoologia Generale e Agraria della R. Scuola Superiore d'Agricoltura in Portici, 5, 285–286.

    Taylor, D.B., Moon, R., Gibson, G. & Szalanski, A. (2006) Genetic and morphological comparisons of new and world populations of Spalangia Species (Hymenoptera: Pteromalidade). Annals of the Entomological Society of America, 99, 799–808.[799:GAMCON]2.0.CO;2

    Uvarov, B. (1966) Grasshoppers and Locusts. Vol. I. Cambridge University Press, Cambridge, pp.

    Veloso, V.R.S., Ferreira, G.A., Fernandes, P.M., Canal Daza, N.A. & Zucchi, R.A. (1996) Ocorrência e índice de infestação de Anastrepha spp. (Dip., Tephritidae) em Pouteria gardneriana Radlk. e Pouteria ramiflora (Mart.) Radlk. (Sapotaceae), nos cerrados de Goiás. Anais das escolas de agronomia e veterinaria, 26, 109–120.

    Viereck, H.L. (1911) Descriptions of six new genera and thirty-one new species of Ichneumon flies. Proceedings of the United States National Museum, 40, 173–196.

    Villemant, C., Simbolotti, G. & Kenis, M. (2007) Discrimination of Eubazus (Hymenoptera, Braconidae) sibling species using geometric morphometrics analysis of wing venation. Systematic Entomology, 32, 625–634.

    Walker, A.K. & Wharton, R.A. (2011) A review of New World Eurytenes s. str. (Hymenoptera, Braconidae, Opiinae). Journal of Hymenoptera Research, 20, 23–46.

    Weekers, P.H.H., De Jonckheere, J.F. & Dumont, H. (2001) Phylogenetic relationships inferred from Ribosomal ITS Sequences and Biogeographic patterns in Representatives of the genus Calopteryx (Insecta: Odonata) of the West Mediterranean and adjacent West European Zone. Molecular Phylogenetics and Evolution, 20, 88–99.

    Wharton, R.A. (1997a) Subfamily Opiinae. In: Wharton, R.A., Marsh, P.M. & Sharkey, M.J. (Eds.), Manual of the New World Genera of the Family Braconidae (Hymenoptera). Allen Press, Lawrence, KS, pp. 379–390.

    Wharton, R.A. (1997b) Generic relationships of Opiinae Braconidae (Hymenoptera) parasitic on fruit-infesting Tephritidae (Diptera). Contributions of the American Entomological Institute, Ann Arbor, 30, 1–53.

    Whitman, D.W. & Agrawal, A.A. (2009) What is phenotypic plasticity and why is it important? In: Whitman, D.W. & Ananthakrishnan, T.N. (Eds.), Phenotypic Plasticity of Insects: Mechanisms and Consequences. Science Publishers, Enfield, NH, pp. 1–63.

    Yoder, M.J., Mikó, I., Seltmann, K., Bertone, M.A. & Deans, A.R. (2010) A gross anatomy ontology for Hymenoptera. PLos ONE, 5 (12), e15991.

    Zucchi, R.A. (2002) A taxonomia e o controle biológico de pragas. In: Parra, J.P., Botelho, P.S., Corrêa-Ferreira, B.S. & Bento, J.M.S. (Eds.), Controle Biológico no Brasil: Parasitóides e predadores. Manole Press, São Paulo, SP, pp. 17–24.

    Zucchi, R.A., Marinho, C.F. & Adaime, R. (2011) First record of the fruit fly parasitoid Doryctobracon crawfordi (Viereck) (Hymenoptera: Braconidae) in Brazil. Neotropical Entomology, 40, 711–712.