Journal of Molecular Evolution

, Volume 58, Issue 6, pp 754–761

Genetic Divergence in the cacophony IVS6 Intron Among Five BrazilianPopulations of Lutzomyia longipalpis

  • M. Bottecchia
  • S. G. Oliveira
  • L. G. S. R. Bauzer
  • N. A. Souza
  • R. D. Ward
  • K. J. Garner
  • C. P. Kyriacou
  • A. A. Peixoto


Genes involved in the reproductive isolation are particularly useful as molecular markers in speciation studies. Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae), a putative species complex, is a vector of visceral leishmaniasis in Latin America. We isolated from this species a fragment homologous to cacophony, a Drosophila gene that encodes features of the lovesong, an acoustic signal that is important in the sexual isolation of closely related species and known to vary considerably among L. longipalpis putative siblings species. Using an intron of the sandfly cacophony as a marker, we analyzed the molecular variation and sequence divergence among five populations of L. longipalpis from Brazil, three allopatric (Jacobina, Lapinha and Natal) and two putative sympatric sibling species from the locality of Sobral. A high level of polymorphism was found and analysis of the data indicates that very little gene flow is occurring among the populations of Jacobina, Lapinha, and Natal. A high level of differentiation was also observed between the two putative sympatric species of Sobral, one of which seems to be the same sibling species found in Natal, while the other is somewhat more related to Jacobina and Lapinha. However, the amount of estimated gene flow among the Sobral siblings is about seven times higher than the previously estimated for period, another lovesong gene, perhaps indicating that introgression might be affecting cacophony more than period. The results suggest that L. longipalpis is not a single species in Brazil, but it is yet not clear whether the different populations studied deserve species status rather than representing an incipient speciation process.


Lutzomyia longipalpis cacophony Speciation Courtship song Leishmaniasis 


  1. Alt, S, Ringo, J, Talyn, B, Bray, W, Dowse, H 1998The period gene controls courtship song cycles in Drosophila melanogasterAnim Behav568797CrossRefPubMedGoogle Scholar
  2. Arrivillaga, JC, Feliciangeli, MD 2001Lutzomyia pseudolongipalpis: The first new species within the longipalpis (Diptera: Psychodidae: Phlebotominae) complex from La Rinconada, Curarigua, Lara State, VenezuelaJ Med Entomol38783790PubMedGoogle Scholar
  3. Arrivillaga, JC, Norris, DE, Feliciangeli, MD, Lanzaro, GC 2002Phylogeography of the neotropical sand fly Lutzomyia longipalpis inferred from mitochondrial DNA sequencesInf Gen Evol28395CrossRefGoogle Scholar
  4. Azevedo, ACR, Monteiro, FA, Cabello, PH, Souza, NA, Rosa-Freitas, MG, Rangel, EF 2000Studies on populations of Lutzomyia longipalpis (Lutz. & Neiva, 1912) (Diptera: Psychodidae: Phlebotominae) in BrazilMem Inst Oswaldo Cruz95305322CrossRefPubMedGoogle Scholar
  5. Bauzer, LGSR, Souza, NA, Ward, RD, Kyriacou, CP, Peixoto, AA 2002aThe period gene and genetic differentiation between three Brazilian populations of Lutzomyia longipalpisInsect Mol Biol11315323CrossRefGoogle Scholar
  6. Bauzer, LGSR, Gesto, JSM, Souza, NA, Ward, RD, Hamilton, JGC, Kyriacou, CP, Peixoto, AA 2002bMolecular divergence in the period gene between two putative sympatric species of the Lutzomyia longipalpis complexMol Biol Evol1916241627Google Scholar
  7. Black, WC, Lanzaro, GC 2001Distribution of genetic variation among chromosomal forms of Anopheles gambiae s.s: Introgressive hybridization, adaptive inversions, or recent reproductive isolation?Insect Mol Biol1037CrossRefPubMedGoogle Scholar
  8. Dujardin, JP, Torrez, EM, Le Pont, F, Hervas, D, Sossa, D 1997Isozymic and metric variation in the Lutzomyia longipalpis complexMed Vet Entomol11394400PubMedGoogle Scholar
  9. Excoffier, L, Smouse, PE, Quattro, JM 1992Analysis of molecular variance inferred from metric distances among DNA haplotypes: Application to human mitochondrial DNA restriction dataGenetics131479491PubMedGoogle Scholar
  10. Filatov, DA, Charlesworth, D 1999DNA polymorphism, haplotype structure and balancing selection in the Leavenworthia PgiC locusGenetics15514231434Google Scholar
  11. Fu, YX, Li, WH 1993Statistical tests of neutrality of mutationsGenetics133693709PubMedGoogle Scholar
  12. Hamilton, JGC, Hooper, AM, Mori, K, Pickett, JA, Sano, S 1999a3-Methyl-α-himachalene confirmed, and the relative stereochemistry defined, by synthesis as the sex pheromone of the sandfly Lutzomyia longipalpis from Jacobina, BrazilChem Commpp 355356CrossRefGoogle Scholar
  13. Hamilton, JGC, Ibbotson, HC, Hooper, AM, Mori, K, Pickett, JA, Sano, S 1999b9-Methylgermacrene-B confirmed by synthesis as the sex pheromone of the sandfly Lutzomyia longipalpis from Lapinha, Brazil, and the absolute stereochemistry defined as 9SChem Comm.23352336CrossRefGoogle Scholar
  14. Hey, J, Wakeley, J 1997A coalescent estimator of the population recombination rateGenetics145833846PubMedGoogle Scholar
  15. Hudson, RR, Kreitman, M, Aguade, M 1987A test of neutral molecular evolution based on nucleotide dataGenetics116153159PubMedGoogle Scholar
  16. Hudson, RR, Slatkin, M, Maddison, WP 1992Estimation of levels of gene flow from DNA sequence dataGenetics132583589PubMedGoogle Scholar
  17. Krzywinski, J, Besansky, NJ 2003Molecular systematics of Anopheles: From subgenera to subpopulationsAnnu Rev Entomol48111139CrossRefPubMedGoogle Scholar
  18. Kumar, S, Tamura, K, Jakobsen, IB, Nei, M 2001MEGA2: Molecular Evolutionary Genetics Analysis softwareBioinformatics1712441245CrossRefPubMedGoogle Scholar
  19. Kyriacou, CP, Hall, JC 1989Spectral-analysis of Drosophila courtship song rhythmsAnim Behav37850859CrossRefGoogle Scholar
  20. Lampo, M, Torgerson, D, Márquez, LM, Rinaldi, M, Garcia, CZ, Arab, A 1999Occurrence of sibling species of Lutzomyia longipalpis (Diptera: Psychodidae) in Venezuela: First evidence from reproductively isolated sympatric populationsAm J Trop Med Hyg6110041009PubMedGoogle Scholar
  21. Lanzaro, GC, Ostrovska, K, Herrero, MV, Lawyer, PG, Warburg, A 1993Lutzomyia longipalpis is a species complex: Genetic divergence and interspecific hybrid sterility among three populationsAm J Trop Med Hyg48839847PubMedGoogle Scholar
  22. Lins, RMMA, Oliveira, SG, Souza, NA, Queiroz, RG, Justiniano, SCB, Ward, RD, Kyriacou, CP, Peixoto, AA 2002Molecular evolution of the cacophony IVS6 region in sandfliesInsect Mol Biol11117122CrossRefPubMedGoogle Scholar
  23. Machado, CA, Kliman, RM, Market, J, Hey, J 2002Inferring the history of speciation from multilocus DNA sequence data: The case of Drosophila pseudoobscura and close relativesMol Biol Evol19472488PubMedGoogle Scholar
  24. Maingon, RDC, Ward, RD, Hamilton, JGC, Noyes, HA, Souza, N, Kemp, SJ, Watts, PC 2003Genetic identification of two sibling species of Lutzomyia longipalpis (Diptera: Psychodidae) that produce distinct male sex pheromones in Sobral, Ceará State, BrazilMol Ecol1218791894CrossRefPubMedGoogle Scholar
  25. Mukhopadhyay, J, Ghosh, K, Rangel, EF, Munstermann, LE 1998aGenetic variability in biochemical characters of Brazilian field populations of the Leishmania vector, Lutzomyia longipalpis (Diptera: Psychodidae)Am J Trop Med Hyg59893901Google Scholar
  26. Mukhopadhyay, J, Ghosh, K, Azevedo, AC, Rangel, EF, Munstermann, LE 1998bGenetic polymorphism of morphological and biochemical characters in a Natal, Brazil, population of Lutzomyia longipalpis (Diptera: Psychodidae)J Am Mosq Control Assoc14277282Google Scholar
  27. Mutebi, JP, Alexander, B, Sherlock, I, Wellington, J, Souza, AA, Shaw, J, Rangel, EF, Lanzaro, GC 1999Breeding structure of the sand fly Lutzomyia longipalpis (Lutz & Neiva) in BrazilAm J Trop Med Hyg61149157Google Scholar
  28. Oliveira, SG, Bottecchia, M, Bauzer, LGSR, Souza, NA, Ward, RD, Kyriacou, CP, Peixoto, AA 2001Courtship song genes and speciation in sandfliesMem Inst Oswaldo Cruz96403405PubMedGoogle Scholar
  29. Peixoto, AA, Hall, JC 1998Analysis of temperature-sensitive mutants reveals new genes involved in the courtship song of DrosophilaGenetics148827838PubMedGoogle Scholar
  30. Peixoto, AA, Smith, LA, Hall, JC 1997Genomic organization and evolution of alternative exons in a Drosophila calcium channel geneGenetics14510031013PubMedGoogle Scholar
  31. Peixoto, AA, Gomes, CA, Amoretty, PR, Lins, RMMA, Meireles-Filho, ACA, Souza, NA, Kyriacou, CP 2001New molecular markers for Phlebotomine sand fliesInt J Parasitol31635639PubMedGoogle Scholar
  32. Rozas, J, Rozas, R 1999DnaSP version 3: An integrated program for population genetics and molecular evolution analysisBioinformatics15174175CrossRefPubMedGoogle Scholar
  33. Saitou, N, Nei, M 1987The neighbor-joining method: A new method for reconstructing phylogenetic treesMol Biol Evol4406425PubMedGoogle Scholar
  34. Schneider, S, Roessli, D, Excoffier, L 2000Arlequin ver. 2.000: A software for population genetics data analysisGenetics and Biometry Laboratory, University of GenevaGeneva, SwitzerlandGoogle Scholar
  35. Sene, FM, Carson, HL 1977Genetic variation in Hawaiian Drosophila. IV. Allozymic similarity between D. silvestris and D. heteroneura from the island of HawaiiGenetics86187198PubMedGoogle Scholar
  36. Smith, LA, Peixoto, AA, Kramer, EM, Villella, A, Hall, JC 1998Courtship and visual defects of cacophony mutants reveal functional complexity of a calcium-channel α-1 subunit in DrosophilaGenetics14914071426PubMedGoogle Scholar
  37. Soto, SI, Lehmann, T, Rowton, ED, Velez, BID, Porter, CH 2001Speciation and population structure in the morphospecies Lutzomyia longipalpis (Lutz & Neiva) as derived from the mitochondrial ND4 geneMol Phylogenet Evol188493PubMedGoogle Scholar
  38. Souza, NA, Ward, RD, Hamilton, JGC, Kyriacou, CP, Peixoto, AA 2002Copulation songs in three siblings of Lutzomyia longipalpis (Diptera:Psychodidae)Trans RR Soc Trop Med Hyg96102103Google Scholar
  39. Tajima, F 1989Statistical method for testing the neutral mutation hypothesis by DNA polymorphismGenetics123585595PubMedGoogle Scholar
  40. Thompson, JD, Gibson, TJ, Plewniak, F, Jeanmougin, F, Higgins, DG 1997The ClustalX windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis toolsNucleic Acids Res2448764882CrossRefGoogle Scholar
  41. Ting, CT, Tsaur, SC, Wu, CI 2000The phylogeny of closely related species as revealed by the genealogy of a speciation gene, OdysseusProc. Natl. Acad. Sci. USA9753135316PubMedGoogle Scholar
  42. Uribe, S 1999The status of the Lutzomyia longipalpis species complex and possible implications for Leishmania transmissionMem Inst Oswaldo Cruz94729734PubMedGoogle Scholar
  43. Wakeley, J, Hey, J 1997Estimating ancestral population parametersGenetics145847855PubMedGoogle Scholar
  44. Wang, RL, Wakeley, J, Hey, J 1997Gene flow and natural selection in the origin of Drosophilapseudoobscura and close relativesGenetics14710911106PubMedGoogle Scholar
  45. Ward, RD, Ribeiro, AL, Ready, PR, Murtagh, A 1983Reproductive isolation between different forms of Lutzomyia longipalpis (Lutz & Neiva), (Diptera: Psychodidae), the vector of Leishmania donovani chagasi Cunha & Chagas and its significance to kala-azar distribution in South AmericaMem Inst Oswaldo Cruz78269280Google Scholar
  46. Ward, RD, Phillips, A, Burnet, B, Marcondes, CB 1988

    The Lutzomyia longipalpis complex: Reproduction and distribution

    Service, MW eds. Biosystematics of hematophagous insectsOxford University PressOxford258269
    Google Scholar
  47. Yin, H, Mutebi, JP, Marriott, S, Lanzaro, GC 1999Metaphase karyotypes and G-banding in sandflies of the Lutzomyia longipalpis complexMed Vet Entomol137277PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • M. Bottecchia
    • 1
  • S. G. Oliveira
    • 1
  • L. G. S. R. Bauzer
    • 1
  • N. A. Souza
    • 2
  • R. D. Ward
    • 3
  • K. J. Garner
    • 4
  • C. P. Kyriacou
    • 4
  • A. A. Peixoto
    • 1
  1. 1.Departamento de Bioquímica e Biologia MolecularFundacão Oswaldo CruzRio de JaneiroBrazil
  2. 2.Departamento de EntomologiaFundação Oswaldo CruzRio de JaneiroBrazil
  3. 3.The Centre for Applied Entomology and ParasitologySchool of Life Sciences, University of KeeleKeeleUK
  4. 4.Department of GeneticsUniversity of LeicesterLeicesterUK

Personalised recommendations