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Molecular Biology Reports

, Volume 39, Issue 3, pp 2375–2383 | Cite as

DNA barcoding of nymphalid butterflies (Nymphalidae: Lepidoptera) from Western Ghats of India

  • S. S. Gaikwad
  • H. V. Ghate
  • S. S. Ghaskadbi
  • M. S. Patole
  • Y. S. Shouche
Article

Abstract

We have checked the utility of DNA barcoding for species identification of nymphalid butterflies from Western Ghats of India by using 650 bp sequence of mitochondrial gene cytochrome c oxidase subunit I. Distinct DNA barcoding gap (i.e. difference between intraspecies and interspecies nucleotide divergence), exists between species studied here. When our sequences were compared with the sequences of the conspecifics submitted from different geographic regions, nine cases of deep intraspecies nucleotide divergences were observed. In spite of this, NJ (Neighbour Joining) clustering analysis successfully discriminated all species. Observed cases of deep intraspecies nucleotide divergences certainly warrant further study.

Keywords

Western Ghats of India DNA barcoding Butterflies Nymphalidae Cytochrome c oxidase I 

Notes

Acknowledgments

We would like to thank the authorities of Modern college, Pune-411005 and The Director, NCCS, Pune-411007 for the facilities provided. We also thank Mr. Parag Ragnekar and Dr. Amol Patwardhan for providing some specimen. This study was supported by DBT (Department of Biotechnology), Govt. of India, Grant No-BT/PR8351/NDB/51/138/2006. SG also thanks DBT for providing fellowship. We would also like to thank the anonymous referees for their valuable comments.

Supplementary material

11033_2011_988_MOESM1_ESM.doc (322 kb)
Supplementary material 1 (DOC 373 kb)

References

  1. 1.
    Blaxter ML (2004) The promise of DNA taxonomy. Philos Trans R Soc Lond Biol Sci 359:669–679CrossRefGoogle Scholar
  2. 2.
    Ratnasingham S, Hebert PDN (2007) BOLD: the barcode of life data system (wwwbarcodinglifeorg). Mol Ecol Notes 7:355–364PubMedCrossRefGoogle Scholar
  3. 3.
    Hebert PDN, Cywinska A, Ball SL, de Waard JR (2003) Biological identification through DNA barcodes. Proc R Soc Lond B Biol Sci 270:313–322CrossRefGoogle Scholar
  4. 4.
    Hebert PDN, Ratnasingham S, deWaard JR (2003b) Barcoding animal life: cytochrome c oxidase subunit 1 divergence among closely related species. Proc R Soc Lond B Biol Sci. doi: 10.1098/rsbl20030025
  5. 5.
    Scheffer SJ, Lewis ML, Joshi RC (2006) DNA barcoding applied to invasive Leafminers (Diptera: Agromyzidae) in the Philippines. Ann Entomol Soc Am 99(2):204–210CrossRefGoogle Scholar
  6. 6.
    Kumar NP, Rajavel AR, Natrajan R, Jambulingam P (2007) DNA barcodes can distinguish species of Indian mosquitoes (Diptera: Culicidae). J Med Entomol 44(1):1–7PubMedCrossRefGoogle Scholar
  7. 7.
    Hastings JM, Schultheis PJ, Whitson M, Holliday CW, Coelho JR, Mendell AM (2008) DNA barcoding of new World cicada killers (Hymenoptera: Crabronidae). Zootaxa 1713:27–38Google Scholar
  8. 8.
    Sinclair CS, Gresens SE (2008) Discrimination of Cricotopus species (Diptera: Chironomidae) by DNA barcoding. Bull Entomol Res 98:555–563PubMedCrossRefGoogle Scholar
  9. 9.
    Ball SL, Hebert PDN, Burian SK, Webb JM (2005) Biological identification of mayflies (Ephemeroptera) using DNA barcodes. J North Am Benthol Soc 24(3):508–524Google Scholar
  10. 10.
    Hogg ID, Hebert PDN (2004) Biological identification of springtails (Hexapoda: Collembola) from the Canadian arctic using mitochondrial DNA barcodes. Can J Zool 82:749–754CrossRefGoogle Scholar
  11. 11.
    Smith MA, Fisher BL, Hebert PDN (2005) DNA barcoding for effective biodiversity assessment of a hyperdiverse arthropod group: the ants of Madagascar. Philos Trans R Soc Lond Biol Sci 360:1825–1834CrossRefGoogle Scholar
  12. 12.
    Johnson SB, Vrijenhoek RC, Waren A (2008) DNA barcoding of Lepetodrilus limpets reveals cryptic species. J Shellfish Res 27(1):43–51CrossRefGoogle Scholar
  13. 13.
    Hou ZE, Li Z, Li SQ (2009) Identifying Chinese species of Gammarus (Crustacea: Amphipoda) using DNA barcoding. Curr Zool 55(2):158–164Google Scholar
  14. 14.
    Robinson EA, Blagoev GA, Hebert PDN, Adamowicz SJ (2009) Prospects for using DNA barcoding to identify spiders in species-rich genera. ZooKeys 16:27–46Google Scholar
  15. 15.
    Hebert PDN, Penton EH, Burns JM, Janzen DH, Hallwachs W (2004) Ten species in one: DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator. Proc Natl Acad Sci USA 101(41):14812–14817PubMedCrossRefGoogle Scholar
  16. 16.
    Hajibabaei M, Janzen DH, Burns JM, Hallwachs W, Remijio ED, Hebert PDN (2006) Wedding biodiversity inventory of a large and complex Lepitodpetra fauna with DNA barcoding. Philos Trans R Soc Lond B Biol Sci 360:1835–1845Google Scholar
  17. 17.
    Monaghan MT, Balke M, Pons J, Vogler AP (2006) Beyond barcodes: complex DNA taxonomy of a South pacific island radition. Proc R Soc Lond B Biol Sci 273:887–893CrossRefGoogle Scholar
  18. 18.
    Monaghan MT et al (2005) DNA based species delineation in tropical beetles using mitochondrial and nuclear markers. Philos Trans R Soc Lond B Biol Sci 360:1925–1933PubMedCrossRefGoogle Scholar
  19. 19.
    Hebert PDN, Stoeckele MY, Lemalak TS, Francis CM (2004) Identification of birds through DNA barcodes. PLoS Biol 2:1657–1663CrossRefGoogle Scholar
  20. 20.
    Kerr CR, Stoeckle MY, Dove CJ, Weight LA, Francis CM, Herbert PDN (2007) Comprehensive DNA barcode coverage of North American birds. Mol Ecol Notes. doi: 10.1111/j.147-8286.2006.01670.x
  21. 21.
    Kerr CR, Lijtmaer DA, Barreira AS, Hebert PDN, Tubaro PL (2009) Probing evolutionary patterns in neotropical birds through DNA barcodes. PLoS ONE 4(1):e4379PubMedCrossRefGoogle Scholar
  22. 22.
    Hubert N, Hanner R, Holm E, Mandrak NE, Taylor E, Burridge M, Watkinson D, Dumont P, Curry A, Bentzen P, Zhang J, April J, Bernatchez L (2008) Identifying Canadian fresh water fishes through DNA barcodes. PLoS ONE 3(6):e2490CrossRefGoogle Scholar
  23. 23.
    Dasmahapatra KK (2006) DNA barcodes: recent success and future prospect. Heredity 97:254–255PubMedCrossRefGoogle Scholar
  24. 24.
    Moritz C, Cicero C (2004) DNA barcoding promise and pitfalls. PLos Biol 2:e354PubMedCrossRefGoogle Scholar
  25. 25.
    Meyer CP, Paulay G (2005) DNA barcoding: error rates based on comprehensive sampling. PloS Biol 3:e422PubMedCrossRefGoogle Scholar
  26. 26.
    Elias M, Hill RI, Willmott KR et al (2007) Limited performance of DNA barcoding in a diverse community of tropical butterflies. Proc R Soc Lond B Biol Sci 274:2881–2889CrossRefGoogle Scholar
  27. 27.
    Wiemers M, Fiedler K (2007) Does the DNA barcoding gap exist?—a case study in blue butterflies (Lepidoptera: Lycaenidae). Front Zool 4:8PubMedCrossRefGoogle Scholar
  28. 28.
    Lukhtanov VA et al (2009) DNA barcoding Central Asian butterflies: increasing geographical dimension does not significantly reduces the success of species identification. Mol Ecol Resour 9:1302–1310PubMedCrossRefGoogle Scholar
  29. 29.
    Hebert PDN, deWaard JR, Landry JF (2008) DNA barcodes for 1/1000 of the animal kingdom. Biol Lett. doi: 10.1098/rsl20090848
  30. 30.
    Kerr KCR, Birks SM, Kalyakin MV, Red’kin YA, Koblik EA, Hebert PDN (2009) Filling the gap-COI barcode resolution in eastern Palearctic birds. Front Zool 6:29PubMedCrossRefGoogle Scholar
  31. 31.
    Bossuyt F, Bocxlaer IV, Biju SD, Loader SP (2009) Toad radiation reveals into-India dispersal as a source of endemism in the Western Ghats-Srilanka biodiversity hotspots. BMC Evol Biol 9:131PubMedCrossRefGoogle Scholar
  32. 32.
    Bossuyt F et al. (2004) Local endemism within the Western Ghats-Srilanka Biodiversity Hotspots. ScienceGoogle Scholar
  33. 33.
    Robin VV, Sinha A, Ramakrishnan U (2010) Ancient geographical gaps and paleoclimate shape the phylogeography of an endemic bird in the sky island of southern India. PloS ONE 5(10):e13321PubMedCrossRefGoogle Scholar
  34. 34.
    Ragupathy S, Newmaster SG, Murugesan M, Balasubramaniam V (2009) DNA barcoding discriminates a new cryptic grass species revealed in an ethanobotany study by the hill tribes of the Western Ghats in Southern India. Mol Ecol Res 9:164–171CrossRefGoogle Scholar
  35. 35.
    Biju SD, Bocxlaer IV, Giri VB, Loader SP, Bossuyt F (2009) Two new endemic genera and a new species of toad (anura: bufonidae) from Western Ghats of India. BMC Res Notes 2:241PubMedCrossRefGoogle Scholar
  36. 36.
    Biju SD, Bossuyt F (2003) New frog family from India reveals an ancient biogeographical link with the Seychelles. Nature 425:711–714PubMedCrossRefGoogle Scholar
  37. 37.
    Prasad V, Farooqui A, Tripathi SKM, Garg R, Thakur B (2009) Evidence of late palaeocene-early eocene equatorial rain forest refugia in southern Western Ghats. India J Biosci 34:771–797Google Scholar
  38. 38.
    Karanth KP (2006) Out of India Gondwana origin of some of tropical Asian biota. Curr Sci 6(90):789–792Google Scholar
  39. 39.
    Datta Roy A, Karanth KP (2009) The Out-of-India hypothesis: what do molecules suggest? J Biosci 34:687–697PubMedCrossRefGoogle Scholar
  40. 40.
    Sechrest W et al (2002) Hotspots and the conservartion of evolutionary history. Proc Natl Acad Sci USA 99(4):2067–2071PubMedCrossRefGoogle Scholar
  41. 41.
    Bickford D et al (2006) Cryptic species as a window on diversity and conservation. Trends Ecol Evol 22(3):148–155PubMedCrossRefGoogle Scholar
  42. 42.
    Gunawardene NR et al (2007) A brief overview of the Western Ghats-Srilanka biodiversity hotspots. Curr Sci 93(11):1567–1572Google Scholar
  43. 43.
    Antram CB (1924) Butterflies of India. Mittal Publication, New DelhiGoogle Scholar
  44. 44.
    Peile HD (1937) A guide to collecting butterflies of India. John Bale Sons & Danielsson Ltd, LondonGoogle Scholar
  45. 45.
    Gunthilagaraj K, Perumal TNA, Jayaram K, Ganesh kumar M (1998) Some South Indian Butterflies Field Guide Published under project lifescape. Indian Academy of Sciences, BangaloreGoogle Scholar
  46. 46.
    Kunte K (2000) Butterflies of peninsular India. Universities Press (India) Limited, HyderabadGoogle Scholar
  47. 47.
    Rangnekar P (2007) The Butterflies of Goa. Mineral Foundation of Goa, GoaGoogle Scholar
  48. 48.
    Casiraghi M, Labra M, Ferri E, Galimberti A, Mattia ED (2010) DNA barcoding: a six-question tour to improve users’ awareness about the method. Brief Bioinform. doi: 10.1093/bib/bbq003
  49. 49.
    Papadopoulou A et al (2008) Speciation and DNA barcodes: testing the effects of dispersal on the formation of discrete sequence clusters. Phil Trans R Soc B 363:2987–2996PubMedCrossRefGoogle Scholar
  50. 50.
    Tyagi A, Bag SK, Shukla V, Roy S, Tuli R (2010) Oligonucleotide frequencies of barcoding loci can discriminate species across kingdoms. PLoS ONE 5(8):e12330PubMedCrossRefGoogle Scholar
  51. 51.
    Frezal L, Leblois R (2008) Four years of DNA barcoding: current advances and prospects. Infect Genet Evol. doi: 10.1016/j.meegid.2008.05.2005
  52. 52.
    Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599PubMedCrossRefGoogle Scholar
  53. 53.
    Sambrook J, Fritsch EF, Manniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold spring Harbor laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  54. 54.
    Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp Ser 41:95–98Google Scholar
  55. 55.
    Xia X, Xie Z (2001) DAMBE: data analysis in molecular biology and evolution. J Hered 92:371–373PubMedCrossRefGoogle Scholar
  56. 56.
    Librado P, Rozas J (2009) DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25:1451–1452PubMedCrossRefGoogle Scholar
  57. 57.
    Kodandaramaiah U, Wahlberg N (2007) Out of Africa origin and dispersal mediated diversification of the butterfly genus Junonia (Nymphalidae: Nymphalinae). J Evol Biol 20:2182–2191CrossRefGoogle Scholar
  58. 58.
    Craft KJ, Pauls SU, Darrow K, Miller SE, Hebert PDN, Helgen LE, Novotny V, Weilen GD (2010) Population genetics of ecological communities with DNA barcodes: an example from New Guinea Lepidoptera. Proc Natl Acad Sci USA 107(11):5041–5046PubMedCrossRefGoogle Scholar
  59. 59.
    Burns JM et al (2008) DNA barcodes and cryptic species of skipper butterflies in the genus Perichares in Area de Conservacion Guanacaste Costarica. Proc Natl Acad Sci USA 105(17):6350–6355PubMedCrossRefGoogle Scholar
  60. 60.
    Pfenninger M, Schwenk K (2007) Cryptic animal species are homogeneously distributed among taxa and biogeographical regions. BMC Evol Biol 7:121–126PubMedCrossRefGoogle Scholar
  61. 61.
    Wahlberg N, Leneven J, Kodandaramaiah U, Pena C, Nylin S, Freitas AVL, Brower AVZ (2009) Nymphalid butterflies diversify following near demise at the Cretaceous/tertiary boundary. Proc Biol Sci 267(1677):4295–4302CrossRefGoogle Scholar
  62. 62.
    Velzen RV (2009) A new hidden species of the Cymothoe caenis—complex (Lepidopetra: Nymphalidae) from Western Africa. Zootaxa 2197:53–63Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • S. S. Gaikwad
    • 1
  • H. V. Ghate
    • 1
  • S. S. Ghaskadbi
    • 2
  • M. S. Patole
    • 3
  • Y. S. Shouche
    • 3
  1. 1.Department of ZoologyModern CollegePuneIndia
  2. 2.Department of ZoologyUniversity of PunePuneIndia
  3. 3.National Center for Cell SciencesPuneIndia

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