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PCR-SSCP and sequence analysis of three Odontotermes spp. (Order: Isoptera; Family: Termitidae) on the basis of partial 16SrRNA gene

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Abstract

Partial 16S gene fragments were amplified by using specific primers in few species/populations of termites of the genus Odontotermes (Isoptera:Termitidae:Macrotermitinae), and the PCR products were subjected to SSCP analysis. Three haplotypes obtained were subjected to sequencing. The sequences obtained were characterized to see the frequencies of each nucleotide bases. High A + T content was observed. The inter-specific pairwise sequence divergence in Odontotermes spp. ranged from 0% to 4.8% across the entire 16S gene fragment. Identical sequences were found between two populations of O. horni. Individuals of different species having Type-I conformational pattern, i.e. O. obesus (-AI) and O. horni (-MI), as well as Type-II of O. obesus (-UII) and O. bhagwatii (-CHII) had no percent diversity. Phylogenetic trees drawn on the basis of distance Neighbour-joining method revealed clustering of individuals according to their genera and families.

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References

  1. Roonwal ML, Chhotani OB (1989) The fauna of India and adjacent countries: Isoptera (Termites). Zoological Survey of India, Calcutta, p 672

    Google Scholar 

  2. Chhotani OB (1997) The fauna of India and the adjacent countries: Isoptera (Termites) (Family: Termitidea). Zoological Survey of India, Calcutta, p 800

    Google Scholar 

  3. Park HC, Orsini JPG, Majer JD, Hobbs RJ (1996) A model of litter harvesting by the Western Australian wheatbelt termite, Drepanotermes tamminensis (Hill), with particular reference to nutrient dynamics. Ecol Res 11(1):69–78. doi:10.1007/BF02347821

    Article  CAS  Google Scholar 

  4. Bignell E (2000) Introduction to symbiosis. In: Abe T, Bignell DE, Higashi M (eds) Termites: evolution, sociality, symbioses. Kluwer, Dordrecht, pp 189–209

    Google Scholar 

  5. Nutting WL (1990) Insecta, Isoptera. In: Dindal DL (ed) Soil biology guide. Wiley, New York, pp 997–1032

    Google Scholar 

  6. Tho TP, Kirton LG (1992) Termites of Peninsular Malaysia. In: Kirton LG (ed) Malayan forest records, vol 36. Forest Research Institute, Kuala Lumpur, Malaysia, p 224

    Google Scholar 

  7. Miura T, Maekawa K, Kitade O, Abe T (1998) Phylogenetic relationships among subfamilies in higher termites (Isoptera: Termitidae) based on mitochondrial COII gene sequences. Ann Entomol Soc Am 91:515–523

    CAS  Google Scholar 

  8. Jenkins TM, Basten CJ, Kresovich S, Forschler BT (1999) Mitochondrial gene sequence questions Reticulitermes species social structure (Isoptera: Rhinotermitidae). Sociobiology 34:161–172

    Google Scholar 

  9. Jenkins TM, Haverty MI, Basten CJ, Nelson LJ, Page M, Forschler BT (2000) Correlation of mitochondrial haplotypes with cuticular hydrocarbon phenotypes of sympatric Reticulitermes species from the southeastern United States. J Chem Ecol 26:1525–1542. doi:10.1023/A:1005548111591

    Article  CAS  Google Scholar 

  10. Jenkins TM, Dean RE, Verkerk R, Forschler BT (2001) Phylogenetic analysis of two mitochondrial genes and one nuclear intron region illuminate European subterranean termite (Isoptera: Rhinotermitidae) gene flow, taxonomy and introduction dynamics. Mol Phylogenet Evol 20:286–293. doi:10.1006/mpev.2001.0966

    Article  PubMed  CAS  Google Scholar 

  11. Marini M, Mantovani B (2002) Molecular relationships among European samples of Reticulitermes (Isoptera: Rhinotermitidae). Mol Phylogenet Evol 22:454–459. doi:10.1006/mpev.2001.1068

    Article  PubMed  CAS  Google Scholar 

  12. Austin JW, Szalanski AL, Uva P, Bagnere`s AG, Kence A (2002) A comparative genetic analysis of the subterranean termite genus Reticulitermes (Isoptera: Rhinotermitidae). Ann Entomol Soc Am 95:753–760. doi:10.1603/0013-8746(2002)095[0753:ACGAOT]2.0.CO;2

    Article  CAS  Google Scholar 

  13. Austin JW, Szalanski AL, Gold RE, Foster BT (2004) Genetic variation and geographical distribution of the subterranean termite genus Reticulitermes in Texas. Ann Entomol Soc Am 97:575–582. doi:10.1603/0013-8746(2004)097[0548:PAOTST]2.0.CO;2

    Article  Google Scholar 

  14. Austin JW, Szalanski AL, Scheffrahn RH, Messenger MT, Dronnet S, Bagnères AG (2005) Genetic evidence for the synonymy of two Reticulitermes species: Reticulitermes flavipes (Kollar) and Reticulitermes santonensis (Feytaud). Ann Entomol Soc Am 98:395–401. doi:10.1603/0013-8746(2005)098[0395:GEFTSO]2.0.CO;2

    Article  CAS  Google Scholar 

  15. Holland MM, Parsons TJ (1999) Mitochondrial DNA sequence analysis, validation and use for forensic casework. Forensic Sci Rev 11:21–50

    Google Scholar 

  16. Avise JC, Arnold J, Ball RM, Bermingham E, Lamb T, Neigel JE, Reeb CA, Saunders NC (1987) Intraspecific phylogeography: the mitochondrial DNA bridge between population genetics and systematics. Annu Rev Ecol Syst 18:489–522

    Google Scholar 

  17. Simon C, Frati F, Beckenback A, Crespi B, Liu H, Flook P (1994) Evolution, weighting, and phylogenetic utility of mitochondrial gene sequences and a compilation of conserved polymerase chain reaction primers. Ann Entomol Soc Am 87:651–701

    CAS  Google Scholar 

  18. Raghava GP, Solanki RJ, Soni V, Agrawal P (2000) Fingerprinting method for phylogenetic classification and identification of microorganisms based on variation in 16S rRNA gene sequences. Biotechniques 29:108–116

    PubMed  CAS  Google Scholar 

  19. Szalanski AL, Austin JW, Nixon J (2004) Establishment of the African termite, Coptotermes sjostedti (Isoptera: Rhinotermitidae), on the Island of Guadeloupe, French West Indies. Ann Entomol Soc Am 97:872–876. doi:10.1603/0013-8746(2004)097[0556:MPABOH]2.0.CO;2

    Article  Google Scholar 

  20. Sambrook J, Fritsh EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbour Laboratory Press, New York

    Google Scholar 

  21. Yukuhiro K, Sezutsu H, Itoh M, Shimizu K, Banno Y (2002) Significant levels of sequence divergence and gene rearrangements have occurred between the mitochondrial genomes of the wild mulberry silkmoth, Bombyx mandarina and its close relative, the domesticated silkmoth, Bombyx mori. Mol Biol Evol 19(8):1385–1389

    PubMed  CAS  Google Scholar 

  22. Vega FJ, Iniesta P, Caldes T, Sanchez A, Lopez JA, de Juan C, Diaz-Rubio E, Torres A, Balibrea JL, Benito M (1997) p53 exon 5 mutations as a prognostic indicator of shortened survival in non-small-cell lung cancer. Br J Cancer 76:44–51

    PubMed  CAS  Google Scholar 

  23. Girman D (1996) The use of PCR-based single-stranded conformation polymorphism analysis (SSCP-PCR) in conservation genetics. In: Smith TB, Wayne RK (eds) Molecular genetic approaches in conservation. Oxford University Press, Oxford, UK, pp 167–182

    Google Scholar 

  24. Ortí G, Hare MP, Avise JC (1997) Detection and isolation of nuclear haplotypes by PCR-SSCP. Mol Ecol 6:575–580. doi:10.1046/j.1365-294X.1997.00212.x

    Article  PubMed  Google Scholar 

  25. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiples sequence alignments through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680. doi:10.1093/nar/22.22.4673

    Article  PubMed  CAS  Google Scholar 

  26. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402. doi:10.1093/nar/25.17.3389

    Article  PubMed  CAS  Google Scholar 

  27. Scheffrahn RH, Krecek J, Szalanski AL, Austin JW (2005) Synonymy of the Neotropical arboreal termites, Nasutitermes corniger and N. costalis (Isoptera: Termitidae), with evidence from morphology, genetics, and biogeography. Ann Entomol Soc Am 98:273–281. doi:10.1603/0013-8746(2005)098[0273:SONATN]2.0.CO;2

    Article  Google Scholar 

  28. Austin JW, Szalanski AL, Messenger MT, McKern JA, Gold RE (2006) Genetic variation and phylogenetics of Reticulitermes (Isoptera: Rhinotermitidae) from the American great plains. Sociobiology 48(2):142–149

    Google Scholar 

  29. Lo N, Kitade O, Miura T, Constantino R, Matsumota T (2004) Molecular phylogeny of Rhinotermitidae. Insectes Soc 51:365–371. doi:10.1007/s00040-004-0759-8

    Article  Google Scholar 

  30. Black WC, Piesman J (1994) Phylogeny of hard- and soft-tick taxa (Acari: Ixodida) based on mitochondrial 16S r DNA sequences. Proc Natl Acad Sci USA 91(7):1034–1038

    Google Scholar 

  31. Kanazawa H, Nouni T, Futai M (1986) Analysis of Escherichia coli mutations of the H+-transporting ATPase: determination of altered sites of the structural gene. Methods Enzymol 126:595–603. doi:10.1016/S0076-6879(86)26062-0

    Article  PubMed  CAS  Google Scholar 

  32. Orita M, Iwahana H, Kanazawa H, Hayashi K, Sekiya T (1989) Detection of polymorphisms of human DNA by gel electrophoresis as single strand conformation polymorphism. Proc Natl Acad Sci USA 86:2766–2770. doi:10.1073/pnas.86.8.2766

    Article  PubMed  CAS  Google Scholar 

  33. Orita M, Suzuki Y, Sekiya T, Hayashi K (1989) Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics 5:874–879. doi:10.1016/0888-7543(89)90129-8

    Article  PubMed  CAS  Google Scholar 

  34. Hayashi K, Yandell DW (1993) How sensitive is PCR-SSCP? Hum Mutat 2:338–346. doi:10.1002/humu.1380020503

    Article  PubMed  CAS  Google Scholar 

  35. Antolin MF, Bosio CF, Cotton J (1999) Intensive linkage mapping in a wasp (Bracon hebetor) and a mosquito (Aedes aegypti) with single-strand conformation polymorphism analysis of random amplified polymorphic DNA markers. Genetics 143:1727–1738

    Google Scholar 

  36. Burt A, Carter DA, Koenig GL, White TJ, Taylor JW (1996) Molecular markers reveal cryptic sex in the human pathogen Coccidioides immitis. Proc Natl Acad Sci USA 93:770–773. doi:10.1073/pnas.93.2.770

    Article  PubMed  CAS  Google Scholar 

  37. Bagley MJ, Medrano JF, Gall GAE (1997) Polymorphic molecular markers from anonymous nuclear DNA for genetic analysis of populations. Mol Ecol 6:309–320. doi:10.1046/j.1365-294X.1997.00192.x

    Article  PubMed  CAS  Google Scholar 

  38. Friesen VL, Congdon BC, Walsh HE, Birt TP (1997) Intron variation in marbled murrelets detected using analysis of single-stranded conformational polymorphism. Mol Ecol 6:1047–1058. doi:10.1046/j.1365-294X.1997.00277.x

    Article  PubMed  CAS  Google Scholar 

  39. Bagley MJ, Gall GAE (1998) Mitochondrial and nuclear sequence variability among populations of rainbow trout (Oncorhynchus mykiss). Mol Ecol 7:945–962. doi:10.1046/j.1365-294x.1998.00413.x

    Article  PubMed  CAS  Google Scholar 

  40. Jaeckel S, Epplen JT, Kauth M (1998) Polymerase chain reaction–single strand conformation polymorphism or how to detect reliably and efficiently each sequence variation in many samples. Electrophoresis 19:3055–3061. doi:10.1002/elps.1150191802

    Article  PubMed  CAS  Google Scholar 

  41. Gasser RB, Woods WG, Blotkamp C (1999) Screening for nucleotide variations in ribosomal DNA arrays of Oesophagostomum bifurcum by polymerase chain reaction-coupled single-strand conformation polymorphism. Electrophoresis 20:1486–1491. doi:10.1002/(SICI)1522-2683(19990601)20:7<1486::AID-ELPS1486>3.0.CO;2-9

    Article  PubMed  CAS  Google Scholar 

  42. Congdon BC, Piatt JF, Martin K, Friesen VL (2000) Mechanisms of population differentiation in marbled murrelets: historical versus contemporary evolutionary processes. Evolution Int J Org Evolution 54:974–986

    CAS  Google Scholar 

  43. Steel DJ, Trewick SA, Wallis GP (2000) Heteroplasmy of mitochondrial DNA in the ophiuroid, Astrobranchion contrictum. J Hered 91:146–149. doi:10.1093/jhered/91.2.146

    Article  PubMed  CAS  Google Scholar 

  44. Trewick SA, Wallis GP, Morgan-Richards M (2000) Sequence data suggest that phylogeographic pattern in the alpine scree weta (Orthoptera, Anostostomatidae) results from late Pliocene vicariance. Mol Ecol 9:657–682. doi:10.1046/j.1365-294x.2000.00905.x

    Article  PubMed  CAS  Google Scholar 

  45. Jenkins TM, Dean RE, Forschle BT (2002) DNA technology, Interstate commerce, and the likely origin of Formosan subterranean termite (Isoptera: Rhinotermitidae) infestation in Atlanta, Georgia. J Econ Entomol 95:381–389

    Article  PubMed  CAS  Google Scholar 

  46. Austin JW, Szalanski AL, Cabrera BJ (2004) A phylogenetic analysis of the subterranean termite family Rhinotermitidae (Isoptera) using the mitochondrial cytochromeoxidase (COII) gene. Ann Entomol Soc Am 97:548–555. doi:10.1603/0013-8746(2004)097[0548:PAOTST]2.0.CO;2

    Article  CAS  Google Scholar 

  47. Austin JW, Szalanski AL, Kard BM (2004) Distribution and genetic variation of Reticulitermes (Isoptera: Rhinotermitidae) in Oklahoma. Fla Entomol 87(2):152–158. doi:10.1653/0015-4040(2004)087[0152:DAGVOR]2.0.CO;2

    Article  Google Scholar 

  48. Luchetti A, Trenta M, Mantovani B, Marini M (2004) Taxonomy and phylogeny of north Mediterranean Reticulitermes termites (Isoptera: Rhinotermitidae): a new insight. Insectes Soc 51:117–122. doi:10.1007/s00040-003-0715-z

    Article  Google Scholar 

  49. Davison DE, Darlington JPEC, Cook CE (2002) Molecular systematics of some Kenyan termites of the genus Odontotermes (Termitidae: Macrotermitinae). Insectes Soc 42(2):138–143

    Google Scholar 

  50. Szalanski AL, Austin JW, Owens CB (2003) Identification of Reticulitermes spp. (Isoptera: Rhinotermitidae) by polymerase chain reaction-restriction fragment length polymorphism. J Econ Entomol 96(5):1514–1519

    Article  PubMed  CAS  Google Scholar 

  51. Austin JW, Szalanski AL, Scheffrahn RH, Messenger MT (2005) Genetic variation of Reticulitermes flavipes (Isoptera: Rhinotermitidae) in North America using the mitochondrial rRNA 16S gene. Ann Entomol Soc Am 98:980–988. doi:10.1603/0013-8746(2005)098[0980:GVORFI]2.0.CO;2

    Article  CAS  Google Scholar 

  52. King SW, Austin JW, Szalanski AL (2007) Use of soldier pronotal width and mitochondrial DNA sequencing to distinguish the subterranean termites, R. flavipes (kollar) and R. Virginicus (banks) Isoptera: Rhinotermitidae), on the Delmarva Peninsula: Delaware, Maryland and Virginia, USA. Entomol News 118:41–48. doi:10.3157/0013-872X(2007)118[41:UOSPWA]2.0.CO;2

    Article  Google Scholar 

  53. Szalanski AL, Austin JW, McKern JA (2008) Genetic diversity of Reticulitermes termites (Isoptera: Rhinotermitidae) from Lake Wedington, Arkansas. Sociobiology 52(1):95–106

    Google Scholar 

  54. Szalanski AL, Austin JWJ, McKern JA, Scheffrahn RH, Owens CB, Messenger MT (2008) Molecular Phylogeography of Reticulitermes (Isoptera: Rhinotermitidae) Termites from Florida. Sociobiology 52(3):619–632

    Google Scholar 

  55. Xiang B, Kochar TD (1991) Comparison of mitochondrial DNA sequences of seven morphospecies of black flies (Diptera). Genome 34:306–311

    Google Scholar 

  56. Kambhampati S (1995) A phylogeny of cockroaches and related insects based on DNA sequence of mitochondrial ribosomal RNA genes. Proc Natl Acad Sci USA 92:2017–2020

    Article  PubMed  CAS  Google Scholar 

  57. Vargol EL, Husseneder C (2009) Biology of subterranean termites: insights from molecular studies of Reticulitermes and Coptotermes. Ann Rev Entomol 54:379–403

    Article  CAS  Google Scholar 

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Acknowledgements

We acknowledge The Council of Scientific and Industrial research (CSIR), New Delhi, for financial support and Zoological Survey of India (ZSI), Kolkata, for species identification.

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Correspondence to Ranbir Chander Sobti.

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Kumari, M., Sharma, V.L., Sodhi, M. et al. PCR-SSCP and sequence analysis of three Odontotermes spp. (Order: Isoptera; Family: Termitidae) on the basis of partial 16SrRNA gene. Mol Cell Biochem 330, 153–162 (2009). https://doi.org/10.1007/s11010-009-0128-z

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