Journal of Insect Conservation

, Volume 20, Issue 5, pp 753–763

A geometric morphometric and microsatellite analyses of Scaptotrigona mexicana and S. pectoralis (Apidae: Meliponini) sheds light on the biodiversity of Mesoamerican stingless bees

  • Miguel Hurtado-Burillo
  • Laura Jara
  • William de Jesús May-Itzá
  • José Javier G. Quezada-Euán
  • Carlos Ruiz
  • Pilar De la Rúa


Geometric morphometrics and molecular methods are effective tools to study the variability of stingless bee populations and species that merit protection given their worldwide decline. Based on previous evidence of cryptic lineages within the Scaptotrigona genus, we tested the existence of multiple evolutionary lineages within the species S. mexicana and we investigated the status of S. pectoralis. By analyzing their population structure, we found differences between the Pacific and Atlantic populations of each of these species, although geometric morphometrics of the wing only confirmed these results in S. mexicana. There was a tendency towards enhanced genetic differentiation over larger distances in the Atlantic populations of both species but not in the Pacific populations. These results revealed a pattern of differentiation among evolutionary units and a specific distribution of genetic diversity within these Scaptotrigona species in Mesoamerica, suggesting the need for future taxonomic revisions, as well as activities aimed at management and conservation.


Scaptotrigona Stingless bees Geometric morphometrics Microsatellites Evolutionary units 


  1. Albores-González ML, García Guerra TG, Durán Olguín L, Aguliar Ayón A (2011) Experiencia de la Unión de Cooperativas Tosepan en el fomento a la cría de las abejas nativas Pitsilnejmej (Scaptotrigona mexicana) in Memorias del VII Seminario Mesaomerciano sobre abejas nativas, pp 95–99 Google Scholar
  2. Álvarez ME, Morrone JJ (2004) Propuesta de áreas para la conservación de aves de México, empleando herramientas panbiogeográficas e índices de complementariedad. Interciencia 29:112–120Google Scholar
  3. Anducho-Reyes MA, Cognato AI, Hayes JL, Zuniga G (2008) Phylogeography of the bark beetle Dendroctonus mexicanus Hopkins (Coleoptera: Curculionidae: Scolytinae). Mol Phylogenet Evol 49:930–940CrossRefPubMedGoogle Scholar
  4. Arias MC, Brito RM, Francisco FO, Moretto G, Oliveira FF, Silvestre D, Sheppard WS (2006) Molecular markers as a tool for population and evolutionary studies of stingless bee. Apidologie 37:259–274CrossRefGoogle Scholar
  5. Ayala R (1999) Revisión de las abejas sin aguijón de México (Hymenoptera, Apoidea). Folia Entomol Mex 106:1–123Google Scholar
  6. Ayala R, González VH, Engels MS (2013) Mexican stingless bees (Hymenoptera: Apidae): diversity, distribution, and indigenous knowledge. In: Vit P, Pedro SRM, Roubik DW (eds) Pot-Honey: a legacy of stingless bees. Springer, New York, pp 135–152CrossRefGoogle Scholar
  7. Balloux F, Lugon-Moulin N (2002) The estimation of population differentiation with microsatellite markers. Mol Ecol 11:155–165CrossRefPubMedGoogle Scholar
  8. Barkan NP, Aytekin AM (2013) Systematical studies on the species of the subgenus Bombus (Thoracobombus) (Hymenoptera: Apidae, Bombus Latreille) in Turkey. Zootaxa 3737(2):167–183CrossRefPubMedGoogle Scholar
  9. Bonatti V, Simões ZLP, Franco FF, Francoy TM (2014) Evidence of at least two evolutionary lineages in Melipona subnitida (Apidae, Meliponini) suggested by mtDNA variability and geometric morphometrics of forewings. Naturwissenschaften 101:17–24CrossRefPubMedGoogle Scholar
  10. Bookstein FL (1991) Morphometric tools for landmark data. Cambridge University Press, CambridgeGoogle Scholar
  11. Borges AA, Campos LAO, Salomão TMF, Tavares MG (2010) Genetic variability in five populations of Partamona helleri (Hymenoptera: Apidae) from Minas Gerais State, Brazil. Genet Mol Biol 33:781–784CrossRefPubMedPubMedCentralGoogle Scholar
  12. Bouga M, Alaux C, Bienkowska M, Büchler R, Carreck NL, Cauia E, Chlebo R, Dahle B, Dall’Olio R, De la Rúa P, Gregorc A, Ivanova E, Kence A, Kence M, Kezic N, Kiprijanovska H, Kozmus P, Kryger P, Le Conte Y, Lodesani M, Murilhas AM, Siceanu A, Soland G, Uzunov A, Wilde J (2011) A review of methods for discrimination of honey bee populations as applied to European beekeeping. J Apic Res 50(1):51–84CrossRefGoogle Scholar
  13. Brown JC, Albrecht C (2001) The effect of tropical deforestation on stingless bees of the genus Melipona (Insecta: Hymenoptera: Apidae: Meliponini) in central Rondonia, Brazil. J Biogeogr 28:623–624CrossRefGoogle Scholar
  14. Brown MJF, Paxton RJ (2009) The conservation of bees: a global perspective. Apidologie 40:410–416CrossRefGoogle Scholar
  15. Byatt MA, Chapman NC, Latty T, Oldroyd BP (2016) The genetic consequences of the anthropogenic movement of social bees. Insectes Soc 63(1):15–24CrossRefGoogle Scholar
  16. Camargo JMF, Pedro SRM (2013) Meliponini Lepeletier, 1836. In: Moure JS, Urban D, Melo GAR. (Orgs). Catalogue of bees (Hymenoptera, Apoidea) in the Neotropical Region-online version; [cited 2012 Sep 29]. Accessed 23 Sept 2015
  17. da Silva FL, Sella MLG, Francoy TM, Costa AHR (2015) Evaluating classification and feature selection techniques for honeybee subspecies identification using wing images. Comput Electron Agric 114:68–77CrossRefGoogle Scholar
  18. Dorn PL, Calderon C, Melgar S, Moguel B, Solorzano E, Dumonteil E, Rodas A, De la Rúa N, Garnica R, Monroy C (2009) Two distinct Triatoma dimidiata (Latreille, 1811) taxa are found in sympatry in Guatemala and Mexico. PLoS Negl Trop Dis 3:e393CrossRefPubMedPubMedCentralGoogle Scholar
  19. Duarte OMP, Gaiotto FA, Costa MA (2014) Genetic differentiation in the stingless bee, Scaptotrigona xanthotricha Moure, 1950 (Apidae, Meliponini): a species with wide geographic distribution in the Atlantic rainforest. J Hered 105(4):477–484CrossRefGoogle Scholar
  20. Earl DA, vonHoldt BM (2012) STRUCTURE HARVESTER: a website and program for visualizing STRUCTURE output and implementing the Evanno method. Conser Genet Res 4(2):359–361CrossRefGoogle Scholar
  21. Engels W, Imperatriz-Fonseca VL (1990) Caste development, reproductive strategies and control of fertility in honey-bees and stingless bees. In: Engels W (ed) Social insects: an evolutionary approach to castes and reproduction. Springer, Berlin, pp 166–230CrossRefGoogle Scholar
  22. Estoup A, Scholl A, Pouvreau A, Solignac M (1995) Monoandry and polyandry in bumble bees (Hymenoptera: Bombinae) as evidenced by highly variable microsatellites. Mol Ecol 4:89–93CrossRefPubMedGoogle Scholar
  23. Evanno G, Regnaut S, Goudet J (2005) Detecting the number of clusters of individuals using the software structure: a simulation study. Mol Ecol 14:2611–2620CrossRefPubMedGoogle Scholar
  24. Excoffier L, Lischer HE (2010) Arlequin suite ver 3.5: a new series of programs to perform population genetics analyses under Linux and Windows. Mol Ecol Res 10(3):564–567CrossRefGoogle Scholar
  25. Fernandes CRM, Martins CF, Ferreira KM, Del Lama MA (2012) Gene variation, population differentiation and sociogenetic structure of nests of Partamona seridoensis (Hymenoptera: Apidae, Meliponini). Biochem Genet 50:325–335CrossRefPubMedGoogle Scholar
  26. Ferreira VS, Aguiar CML, Costa MA, Silva JG (2011) Morphometric analysis of populations of Centris aenea Lepeletier (Hymenoptera: Apidae) from Northeastern Brazil. Neotrop Entomol 40:97–102CrossRefPubMedGoogle Scholar
  27. Foley JA, DeFries R, Asner GP, Barford C, Bonan G, Carpenter SR, Chapin FS, Coe MT, Daily GC, Gibbs HK, Helkowski JH, Holloway T, Howard EA, Kucharik CJ, Monfreda C, Patz JA, Prentice IC, Ramankutty N, Snyder PK (2005) Global consequences of land use. Science 309:570–574CrossRefPubMedGoogle Scholar
  28. Francisco FO, Brito RM, Arias MC (2006) Allele number of heterozigosity for microsatellite loci in different stingless bee species (Hymenoptera: Apidae: Meliponini). Neotrop Entomol 35:638–643CrossRefGoogle Scholar
  29. Francisco FO, Nunes-Silva P, Francoy TM, Wittmann D, Imperatriz-Fonseca VL, Arias MC, Morgan ED (2008) Morphometrical, biochemical and molecular tools for assessing biodiversity. An example in Plebeia remota (Holmberg, 1903) (Apidae, Meliponini). Insectes Soc 55(3):231–237CrossRefGoogle Scholar
  30. Francoy TM, Grassi ML, Imperatriz-Fonseca VL, May-Itzá WdJ, Quezada-Euán JJG (2011) Geometric morphometrics of the wing as a tool for assigning genetic lineages and geographic origin to Melipona beecheii (Hymenoptera: Meliponini). Apidologie 42:499–507CrossRefGoogle Scholar
  31. Frankham R, Ballou JD, Briscoe DA (2002) Introduction to conservation genetics. Cambridge University Press, CambridgeCrossRefGoogle Scholar
  32. Freitas BM, Imperatriz-Fonseca VL, Medina LM, Kleinert AMP, Galetto L, Nates-Parra G, Quezada-Euán JJG (2009) Diversity, threats and conservation of native bees in the Neotropics. Apidologie 40:332–346CrossRefGoogle Scholar
  33. Gonçalves PHP (2010) Análise da variabilidade genética de uma pequena população de Frieseomelitta varia (Hymenoptera, Apidae, Meliponini) por meio de análise do DNA mitocondrial, microssatélites e morfometria geométrica das asas. Dissertation, University of São PauloGoogle Scholar
  34. González-Acereto JA (2012) La importancia de la meliponicultura en México, con énfasis en la Península de Yucatán. Bioagrociencias 5(1):34–41Google Scholar
  35. González-Acereto JA, Quezada-Euá JJG, Medina-Medina LA (2006) New perspectives for stingless beekeeping in the Yucatan: results of an integral program to rescue and promote the activity. J Apicult Res 45(3):234–239CrossRefGoogle Scholar
  36. Hammer Ø, Harper DAT, Ryan PD (2001) PAST: paleontological statistics software package for education and data analysis. Palaeontol Electron 4:1–9Google Scholar
  37. Hedrick PW (2005) A standardized genetic differentiation measure. Evolution 59:1633–1638CrossRefPubMedGoogle Scholar
  38. Hurtado-Burillo M, Ruiz C, May-Itzá WdJ, Quezada-Eúan JJG, De la Rúa P (2013) Barcoding stingless bees: genetic diversity of the economically important genus Scaptotrigona in Mesoamerica. Apidologie 44:1–10CrossRefGoogle Scholar
  39. Ish-Am G, Barrientos-Priego F, Castañeda-Vildozola A, Gazit S (1999) Avocado (Persea americana Mill) pollinators in its region of origin. Rev Chapingo Ser Hortic 5:137–143Google Scholar
  40. Jaffé R, Pope N, Giannini TC, Acosta AL, Alves DA, Arias MC, Francisco FO, González A, Imperatriz-Fonseca VL, Jha S, De la Rúa P, Tavares MG, Carvalheiro LG (2016) Gene flow in stingless bees is determined by geographic distance and beekeeping practices. Mol Ecol (in press)Google Scholar
  41. Jakobsson M, Rosenberg NA (2007) CLUMPP: a cluster matching and permutation program for dealing with label switching and multimodality in analysis of population structure. Bioinformatics 23:1801–1806CrossRefPubMedGoogle Scholar
  42. Kalinowski ST (2005) HP-Rare: a computer program for performing rarefaction on measures of allelic diversity. Mol Ecol Notes 5:187–189CrossRefGoogle Scholar
  43. Klingenberg CP (2011) MorphoJ: an integrated software package for geometric morphometrics. Mol Ecol Resour 11:353–357CrossRefPubMedGoogle Scholar
  44. Kraus FB, Weinhold S, Moritz RFA (2008) Genetic structure of drone congregations of the stingless bee Scaptotrigona mexicana. Insectes Soc 55:22–27CrossRefGoogle Scholar
  45. Leberg PL (2002) Estimating allelic richness: effects of sample size and bottlenecks. Mol Ecol 11:2445–2449CrossRefPubMedGoogle Scholar
  46. Lima Junior CA, Carvalho CAL, Nunes LA, Francoy TM (2012) Population divergence of Melipona scutellaris Latreille (Hymenoptera: Meliponina) in two Restricted Areas in Bahia, Brazil. Sociobiology 59:107–122CrossRefGoogle Scholar
  47. Luikart G, England PR (1999) Statistical analysis of microsatellite DNA data. Trends Ecol Evol 14:253–255CrossRefPubMedGoogle Scholar
  48. Mann HB, Whitney DR (1947) On a test of whether one of two random variables is stochastically larger than the other. Ann Math Stat 18(1):50–60CrossRefGoogle Scholar
  49. Manzo C (2009) Informe Final de Actividades de la Huasteca. Módulo de Abejas sin Aguijón (Scaptotrigona mexicana) Municipio de Cocoxtlán S.L.PGoogle Scholar
  50. May-Itzá WdJ, Quezada-Eúan JJG, Medina-Medina LA, Enriquez E, De la Rúa P (2010) Morphometric and genetic differentiation in isolated populations of the endangered Mesoamerican stingless bee Melipona yucatanica (Hymenoptera: Apoidea) suggest the existence of a two species complex. Conserv Genet 11:2079–2084CrossRefGoogle Scholar
  51. May-Itzá WdJ, Quezada-Eúan JJG, Ayala R, De la Rúa P (2012) Morphometric and genetic analyses differentiate Mesoamerican populations of the endangered stingless bee Melipona beecheii (Hymenoptera: Meliponidae) and support their conservation as two separate units. J Insect Conserv 16(5):723–731CrossRefGoogle Scholar
  52. Mendes MFM, Francoy TM, Nunes-Silva P, Menezes C, Imperatriz-Fonseca VL (2007) Intra-populational variability of Nannotrigona testaceicornis Lepeletier, 1836 (Hymenoptera, Meliponini) using relative warp analysis. Biosci J 23:147–152Google Scholar
  53. Michener CD (2007) The bees of the world, 2nd edn. The Johns Hopkins University Press, BaltimoreGoogle Scholar
  54. Morandin LA, Winston ML (2006) Pollinators provide economic incentive to preserve natural land in agroecosystems. Agric Ecosyst Environ 116:289–292CrossRefGoogle Scholar
  55. Mueller MY, Moritz RFA, Kraus FB (2012) Outbreeding and lack of temporal genetic structure in a drone congregation of the Neotropical stingless bee Scaptotrigona mexicana. Ecol Evol 2:1304–1311CrossRefPubMedPubMedCentralGoogle Scholar
  56. Nunes LA, Passos GB, Carvalho CAL, Araújo ED (2013) Spatial variation of size and shape of the wing in Melipona quadrifasciata anthidioides Lepeletier, 1836 (Hymenoptera; Meliponini) assessed by geometric morphometrics. Braz J Biol 73:887–893CrossRefPubMedGoogle Scholar
  57. Oleksa A, Tofilski A (2015) Wing geometric morphometrics and microsatellite analysis provide similar discrimination of honey bee subspecies. Apidologie 46(1):49–60CrossRefGoogle Scholar
  58. Paxton RJ, Weibschuh N, Quezada-Euán JJG (1999) Characterization of dinucleotide microsatellite loci for stingless bees. Mol Ecol 8:685–702CrossRefGoogle Scholar
  59. Peakall R, Smouse PE (2006) GENEALEX 6: genetic analysis in Excel, Population genetic software for teaching and research. Mol Ecol Notes 6:288–295CrossRefGoogle Scholar
  60. Peters JM, Queller DC, Imperatriz-Fonseca VL, Strassmann JE (1998) Microsatellite loci for stingless bees. Mol Ecol 7:783–792CrossRefGoogle Scholar
  61. Pritchard JK, Stephens M, Donnelly P (2000) Inference of population structure using multilocus genotype data. Genetics 155:945–959PubMedPubMedCentralGoogle Scholar
  62. Quezada-Euán JJG, May-Itzá WdJ, Rincón M, De la Rúa P, Paxton RJ (2012) Genetic and phenotypic differentiation in endemic Scaptotrigona hellwegeri (Apidae: Meliponini): implications for the conservation of stingless bee populations in contrasting environments. Insect Conserv Divers 5(6):433–443CrossRefGoogle Scholar
  63. Raymond M, Rousset F (1995a) GENEPOP (version 1.2): population genetics software for exact tests and ecumenicism. J Hered 86:248–249Google Scholar
  64. Raymond M, Rousset F (1995b) An exact test for population differentiation. Evolution 49:1283–1286CrossRefGoogle Scholar
  65. Reed DH, Frankham R (2001) How closely correlated are molecular and quantitative measures of genetic variation? A meta-analysis. Evolution 55:1095–1103CrossRefPubMedGoogle Scholar
  66. Rice W (1989) Analysing tables of statistical tests. Evolution 43:223–225CrossRefGoogle Scholar
  67. Rohlf FJ (2013) TPSdig v. 2.17. NY State University at Stony Brook, Stony BrookGoogle Scholar
  68. Rosenberg NA (2004) Distruct: a program for the graphical display of population structure. Mol Ecol Notes 4:137–138CrossRefGoogle Scholar
  69. Russell KN, Ikerd H, Droege S (2005) The potential conservation value of unmowed powerline strips for native bees. Biol Conserv 124:133–148CrossRefGoogle Scholar
  70. Santiago LR, Francisco FO, Jaffé R, Arias MC (2016) Genetic variability in captive populations of the stingless bee Tetragonisca angustula. Genetica 144:397–405CrossRefPubMedGoogle Scholar
  71. Schlick-Steiner BC, Steiner FM, Seifert B, Stauffer C, Christian E, Crozier RH (2010) Integrative taxonomy: a multisource approach to exploring biodiversity. Annu Rev Entomol 55:421–438CrossRefPubMedGoogle Scholar
  72. Slaa EJ, Sánchez LA, Malagodi-Braga KS, Hofstede FE (2006) Stingless bees in applied pollination: practice and perspectives. Apidologie 37:293–315CrossRefGoogle Scholar
  73. Stout JC, Morales CL (2009) Ecological impacts of invasive alien species on bees. Apidologie 40:388–409CrossRefGoogle Scholar
  74. Tavares MG, Dias LAD, Borges AA, Lopes DM, Busse AHP, Costa RG, Salomão TMF, Campos LAO (2007) Genetic divergence between populations of the stingless bee uruçu amarela (Melipona rufiventris group, Hymenoptera, Meliponini): is there a new Melipona species in the Brazilian state of Minas Gerais? Gen Mol Biol 30:667–675CrossRefGoogle Scholar
  75. Valdovinos-Núñez GR, Quezada-Euán JJG, Ancona-Xiu P, Moo-Valle H, Carmona A, Ruiz Sánchez E (2009) Comparative toxicity of pesticides to stingless bees (Hymenoptera: Apidae: Meliponini). J Econ Entomol 102:1737–1742CrossRefPubMedGoogle Scholar
  76. Van Oosterhout C, Hutchinson WF, Wills DP, Shipley P (2004) MICRO-CHECKER: software for identifying and correcting genotyping errors in microsatellite data. Mol Ecol Notes 4:535–538CrossRefGoogle Scholar
  77. Venturieri GC (2009) The impact of forest exploitation on Amazonian stingless bees (Apidae, Meliponini). Genet Mol Res 8:684–689CrossRefPubMedGoogle Scholar
  78. Yáñez-Ordóñez O, Trujano Ortega A, Llorente Bousquets J (2008) Patrones de distribución de las especies de la tribu Meliponini (Hymenoptera: Apoidea: Apidae) en México. Interciencia 33(1):41–45Google Scholar
  79. Zayed A (2009) Bee genetics and conservation. Apidologie 40:237–262CrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2016

Authors and Affiliations

  1. 1.Departamento de Zoología y Antropología Física, Facultad de VeterinariaUniversidad de MurciaMurciaSpain
  2. 2.Departamento de Apicultura, Facultad de Medicina Veterinaria y ZootecniaUniversidad Autónoma de YucatánMéridaMexico

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