Skip to main content
Log in

Characterisation of the complete mitochondrial genome and 13 microsatellite loci through next-generation sequencing for the New Caledonian spider-ant Leptomyrmex pallens

  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

The complete mitochondrial genome and a set of polymorphic microsatellite markers were identified by 454 pyrosequencing (1/16th of a plate) for the New Caledonian rainforest spider-ant Leptomyrmex pallens. De novo genome assembly recovered the entire mitochondrial genome with mean coverage of 8.9-fold (range 1–27). The mitogenome consists of 15,591 base pairs including 13 protein-coding genes, 2 ribosomal subunit genes, 22 transfer RNAs, and a non-coding AT-rich region. The genome arrangement is typical of insect taxa and very similar to the only other published ant mitogenome from the Solenopsis genus, with the main differences consisting of translocations and inversions of tRNAs. A total of 13 polymorphic loci were also characterized using 41 individuals from a single population in the Aoupinié region, corresponding to workers from 21 nests and 16 foraging workers. We observed moderate genetic variation across most loci (mean number of alleles per locus = 4.50; mean expected heterozygosity = 0.53) with evidence of only two loci deviating significantly from Hardy–Weinberg equilibrium due to null alleles. Marker independence was confirmed with tests for linkage disequilibrium. Most loci cross amplified for three additional Leptomyrmex species. The annotation of the mitogenome and characterization of microsatellite markers will provide useful tools for assessing the colony structure, population genetic patterns, and dispersal strategy of L. pallens in the context of rainforest fragmentation in New Caledonia. Furthermore, this paper confirms a recent line of evidence that comprehensive mitochondrial data can be obtained relatively easily from small next-generation sequencing analyses. Greater synthesis of next-generation sequencing data will play a significant role in expanding the taxonomic representation of mitochondrial genome sequences.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Andersen AN, Majer J (2004) Ants show the way Down Under: invertebrates as bioindicators in land management. Front Ecol Environ 2(6):291–298. doi:10.1890/1540-9295(2004)002[0292:astwdu]2.0.co;2

  2. Laurance WF, Lovejoy TE, Vasconcelos HL, Bruna EM, Didham RK, Stouffer PC, Gascon C, Bierregaard RO, Laurance SG, Sampaio E (2002) Ecosystem decay of Amazonian forest fragments: a 22-year investigation. Conserv Biol 16:605–618

    Article  Google Scholar 

  3. Bickel TO, Brühl CA, Gadau JR, Hölldobler B, Linsenmair KE (2006) Influence of habitat fragmentation on the genetic variability in leaf litter ant populations in tropical rainforests of Sabah, Borneo. Biodivers Conserv 15(1):157–175. doi:10.1007/s10531-004-4248-1

    Article  Google Scholar 

  4. Tawato N, Harper NE, Mohamed M, Khen CV, Searle JB, Hill JK (2011) Impacts of forest fragmentation on the genetic diversity and population structure of Pachycondyla obscurans in Sabah, Malaysian Borneo. Asian Myrmecol 4:59–68

    Google Scholar 

  5. Pamilo P, Gertsch P, Thoren P, Seppa P (1997) Molecular population genetics of social insects. Annu Rev Ecol Syst 28:1–25

    Article  Google Scholar 

  6. Crozier RH, Oldroyd BP, Tay WT, Kaufmann BE, Johnson RN, Carew ME, Jennings KM (1997) Molecular advances in understanding social insect population structure. Electrophoresis 18(9):1672–1675. doi:10.1002/elps.1150180934

    Article  CAS  PubMed  Google Scholar 

  7. Gardner MG, Fitch AJ, Bertozzi T, Lowe AJ (2011) Rise of the machines—recommendations for ecologists when using next generation sequencing for microsatellite development. Mol Ecol Resour 11(6):1093–1101. doi:10.1111/j.1755-0998.2011.03037.x

    Article  PubMed  Google Scholar 

  8. Miller A, Good R, Coleman R, Lancaster M, Weeks A (2012) Microsatellite loci and the complete mitochondrial DNA sequence characterized through next generation sequencing and de novo genome assembly for the critically endangered orange-bellied parrot Neophema chrysogaster. Mol Biol Rep 40(1):35–42. doi:10.1007/s11033-012-1950-z

    Article  PubMed  Google Scholar 

  9. Prosdocimi F, Carvalho D, Almeida R, Beheregaray L (2012) The complete mitochondrial genome of two recently derived species of the fish genus Nannoperca (Perciformes, Percichthyidae). Mol Biol Rep 39(3):2767–2772. doi:10.1007/s11033-011-1034-5

    Article  CAS  PubMed  Google Scholar 

  10. Avise JC (2000) Phylogeography: the history and formation of species. President and Fellows of Harvard College, United States of America

  11. Li J, Heinz KM (2000) Genome complexity and organization in the red imported fire ant Solenopsis invicta Buren. Genet Res 75(02):129–135

    Article  CAS  PubMed  Google Scholar 

  12. Wurm Y, Wang J, Riba-Grognuz O, Corona M, Nygaard S, Hunt BG, Ingram KK, Falquet L, Nipitwattanaphon M, Gotzek D, Dijkstra MB, Oettler J, Comtesse F, Shih C-J, Wu W-J, Yang C–C, Thomas J, Beaudoing E, Pradervand S, Flegel V, Cook ED, Fabbretti R, Stockinger H, Long L, Farmerie WG, Oakey J, Boomsma JJ, Pamilo P, Yi SV, Heinze J, Goodisman MAD, Farinelli L, Harshman K, Hulo N, Cerutti L, Xenarios I, Shoemaker D, Keller L (2011) The genome of the fire ant Solenopsis invicta. Proc Natl Acad Sci 108(14):5679–5684. doi:10.1073/pnas.1009690108

    Article  CAS  PubMed  Google Scholar 

  13. Smith CD, Zimin A, Holt C, Abouheif E, Benton R, Cash E, Croset V, Currie CR, Elhaik E, Elsik CG, Fave M-J, Fernandes V, Gadau J, Gibson JD, Graur D, Grubbs KJ, Hagen DE, Helmkampf M, Holley J-A, Hu H, Viniegra ASI, Johnson BR, Johnson RM, Khila A, Kim JW, Laird J, Mathis KA, Moeller JA, Muñoz-Torres MC, Murphy MC, Nakamura R, Nigam S, Overson RP, Placek JE, Rajakumar R, Reese JT, Robertson HM, Smith CR, Suarez AV, Suen G, Suhr EL, Tao S, Torres CW, van Wilgenburg E, Viljakainen L, Walden KKO, Wild AL, Yandell M, Yorke JA, Tsutsui ND (2011) Draft genome of the globally widespread and invasive Argentine ant (Linepithema humile). Proc Natl Acad Sci 108(14):5673–5678. doi:10.1073/pnas.1008617108

    Article  CAS  PubMed  Google Scholar 

  14. Smith CR, Smith CD, Robertson HM, Helmkampf M, Zimin A, Yandell M, Holt C, Hu H, Abouheif E, Benton R, Cash E, Croset V, Currie CR, Elhaik E, Elsik CG, Favé M-J, Fernandes V, Gibson JD, Graur D, Gronenberg W, Grubbs KJ, Hagen DE, Viniegra ASI, Johnson BR, Johnson RM, Khila A, Kim JW, Mathis KA, Munoz-Torres MC, Murphy MC, Mustard JA, Nakamura R, Niehuis O, Nigam S, Overson RP, Placek JE, Rajakumar R, Reese JT, Suen G, Tao S, Torres CW, Tsutsui ND, Viljakainen L, Wolschin F, Gadau J (2011) Draft genome of the red harvester ant Pogonomyrmex barbatus. Proc Natl Acad Sci 108(14):5667–5672. doi:10.1073/pnas.1007901108

    Article  CAS  PubMed  Google Scholar 

  15. Suen G, Teiling C, Li L, Holt C, Abouheif E, Bornberg-Bauer E, Bouffard P, Caldera EJ, Cash E, Cavanaugh A, Denas O, Elhaik E, Favé M-J, Gadau J, Gibson JD, Graur D, Grubbs KJ, Hagen DE, Harkins TT, Helmkampf M, Hu H, Johnson BR, Kim J, Marsh SE, Moeller JA, Muñoz-Torres MC, Murphy MC, Naughton MC, Nigam S, Overson R, Rajakumar R, Reese JT, Scott JJ, Smith CR, Tao S, Tsutsui ND, Viljakainen L, Wissler L, Yandell MD, Zimmer F, Taylor J, Slater SC, Clifton SW, Warren WC, Elsik CG, Smith CD, Weinstock GM, Gerardo NM, Currie CR (2011) The genome sequence of the leaf-cutter ant Atta cephalotes reveals insights into its obligate symbiotic lifestyle. PLoS Genet 7(2):e1002007. doi:10.1371/journal.pgen.1002007

    Article  PubMed Central  PubMed  Google Scholar 

  16. Nygaard S, Zhang G, Schiøtt M, Li C, Wurm Y, Hu H, Zhou J, Ji L, Qiu F, Rasmussen M, Pan H, Hauser F, Krogh A, Grimmelikhuijzen CJP, Wang J, Boomsma JJ (2011) The genome of the leaf-cutting ant Acromyrmex echinatior suggests key adaptations to advanced social life and fungus farming. Genome Res 21(8):1339–1348. doi:10.1101/gr.121392.111

    Article  CAS  PubMed  Google Scholar 

  17. Bonasio R, Zhang G, Ye C, Mutti NS, Fang X, Qin N, Donahue G, Yang P, Li Q, Li C, Zhang P, Huang Z, Berger SL, Reinberg D, Wang J, Liebig J (2010) Genomic comparison of the ants Camponotus floridanus and Harpegnathos saltator. Science 329(5995):1068–1071. doi:10.1126/science.1192428

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  18. Gotzek D, Clarke J, Shoemaker D (2010) Mitochondrial genome evolution in fire ants (Hymenoptera: Formicidae). BMC Evol Biol 10(1):300

    Article  PubMed Central  PubMed  Google Scholar 

  19. Lucky A, Ward PS (2010) Taxonomic revision of the ant genus Leptomyrmex Mayr (Hymenoptera: Formicidae). Zootaxa 2688:1–67

    Google Scholar 

  20. Jaffré T, Bouchet P, Veillon J-M (1998) Threatened plants of New Caledonia: is the system of protected areas adequate? Biodivers Conserv 7:109–135

    Article  Google Scholar 

  21. Margulies M, Egholm M, Altman WE, Attiya S, Bader JS, Bemben LA, Berka J, Braverman MS, Chen Y-J, Chen Z, Dewell SB, Du L, Fierro JM, Gomes XV, Godwin BC, He W, Helgesen S, Ho CH, Irzyk GP, Jando SC, Alenquer MLI, Jarvie TP, Jirage KB, Kim J-B, Knight JR, Lanza JR, Leamon JH, Lefkowitz SM, Lei M, Li J, Lohman KL, Lu H, Makhijani VB, McDade KE, McKenna MP, Myers EW, Nickerson E, Nobile JR, Plant R, Puc BP, Ronan MT, Roth GT, Sarkis GJ, Simons JF, Simpson JW, Srinivasan M, Tartaro KR, Tomasz A, Vogt KA, Volkmer GA, Wang SH, Wang Y, Weiner MP, Yu P, Begley RF, Rothberg JM (2005) Genome sequencing in microfabricated high-density picolitre reactors. Nature 437(7057):376–380. doi:http://www.nature.com/nature/journal/v437/n7057/suppinfo/nature03959_S1.html

    Google Scholar 

  22. Meglécz E, Costedoat C, Dubut V, Gilles A, Malausa T, Pech N, Martin J-F (2010) QDD: a user-friendly program to select microsatellite markers and design primers from large sequencing projects. Bioinformatics 26(3):403–404. doi:10.1093/bioinformatics/btp670

    Article  PubMed  Google Scholar 

  23. Rozen S, Skaletsky H (1999) Primer3 on the WWW for general users and for biologist programmers. In: Misener S, Krawetz S (eds) Bioinformatics methods and protocols. Methods in molecular biology, vol 132. Humana Press, Totowa, pp 365–386. doi:10.1385/1-59259-192-2:365

  24. Blacket MJ, Robin C, Good RT, Lee SF, Miller AD (2012) Universal primers for fluorescent labelling of PCR fragments—an efficient and cost-effective approach to genotyping by fluorescence. Mol Ecol Resour 12(3):456–463. doi:10.1111/j.1755-0998.2011.03104.x

    Article  CAS  PubMed  Google Scholar 

  25. Park S (2001) The Excel microsatellite toolkit. Website http://animalgenomics.ucd.ie/sdepark/ms-toolkit/. Accessed Aug 2007

  26. Raymond M, Rousset F (1995) Genepop (version 1.2): population genetics software for exact tests and ecumenicism. J Hered 86:248–249

    Google Scholar 

  27. Rice WR (1989) Analyzing tables of statistical tests. Evolution 43(1):223–225

    Article  Google Scholar 

  28. Van Oosterhout C, Hutchinson WF, Wills DPM, Shipley P (2004) Micro-checker: software for identifying and correcting genotyping errors in microsatellite data. Mol Ecol Notes 4(3):535–538. doi:10.1111/j.1471-8286.2004.00684.x

    Article  Google Scholar 

  29. Brookfield J (1996) A simple new method for estimating null allele frequency from heterozygote deficiency. Mol Ecol 5(3):453–455

    Article  CAS  PubMed  Google Scholar 

  30. Giardine B, Riemer C, Hardison RC, Burhans R, Elnitski L, Shah P, Zhang Y, Blankenberg D, Albert I, Taylor J, Miller W, Kent WJ, Nekrutenko A (2005) Galaxy: a platform for interactive large-scale genome analysis. Genome Res 15(10):1451–1455. doi:10.1101/gr.4086505

    Article  CAS  PubMed  Google Scholar 

  31. Chevreux B, Pfisterer T, Drescher B, Driesel AJ, Müller WEG, Wetter T, Suhai S (2004) Using the miraEST assembler for reliable and automated mRNA transcript assembly and SNP detection in sequenced ESTs. Genome Res 14(6):1147–1159. doi:10.1101/gr.1917404

    Article  CAS  PubMed  Google Scholar 

  32. Drummond A, Ashton B, Buxton S, Cheung M, Cooper A, Duran C, Field M, Heled J, Kearse M, Markowitz S, Moir R, Stones-Havas S, Sturrock S, Thierer T, Wilson A (2011) Geneious. 5.5.6 edn. Available from http://www.geneious.com/

  33. Eddy SR, Durbin R (1994) RNA sequence analysis using covariance models. Nucleic Acids Res 22(11):2079–2088. doi:10.1093/nar/22.11.2079

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  34. Wyman SK, Jansen RK, Boore JL (2004) Automatic annotation of organellar genomes with DOGMA. Bioinformatics (Oxford, England) 20(17):3252–3255. doi:10.1093/bioinformatics/bth352

  35. Cameron SL, Dowton M, Castro LR, Ruberu K, Whiting MF, Austin AD, Diement K, Stevens J (2008) Mitochondrial genome organization and phylogeny of two vespid wasps. Genome 51(10):800–808. doi:10.1139/g08-066

    Article  CAS  PubMed  Google Scholar 

  36. Crozier RH, Crozier YC (1993) The mitochondrial genome of the honeybee Apis mellifera: complete sequence and genome organization. Genetics 133(1):97–117

    CAS  PubMed  Google Scholar 

  37. Lewis OL, Farr CL, Kaguni LS (1995) Drosophila melanogaster mitochondrial DNA: completion of the nucleotide sequence and evolutionary comparisons. Insect Mol Biol 4(4):263–278. doi:10.1111/j.1365-2583.1995.tb00032.x

    Article  CAS  PubMed  Google Scholar 

  38. Martins J, Solomon SE, Mikheyev AS, Mueller UG, Ortiz A, Bacci M (2007) Nuclear mitochondrial-like sequences in ants: evidence from Atta cephalotes (Formicidae: Attini). Insect Mol Biol 16(6):777–784. doi:10.1111/j.1365-2583.2007.00771.x

    Article  CAS  PubMed  Google Scholar 

  39. Clary D, Wolstenholme D (1985) The mitochondrial DNA molecule of Drosophila yakuba: nucleotide sequence, gene organization, and genetic code. J Mol Evol 22(3):252–271. doi:10.1007/bf02099755

    Article  CAS  PubMed  Google Scholar 

  40. Bae JS, Kim I, Sohn HD, Jin BR (2004) The mitochondrial genome of the firefly, Pyrocoelia rufa: complete DNA sequence, genome organization, and phylogenetic analysis with other insects. Mol Phylogenet Evol 32(3):978–985. doi:10.1016/j.ympev.2004.03.009

    Article  CAS  PubMed  Google Scholar 

  41. Wolstenholme DR (1992) Animal mitochondrial DNA: structure and evolution. Int Rev Cytol 141:173–216

    Article  CAS  PubMed  Google Scholar 

  42. Dowton M, Cameron SL, Dowavic JI, Austin AD, Whiting MF (2009) Characterization of 67 mitochondrial tRNA gene rearrangements in the Hymenoptera suggests that mitochondrial tRNA gene position is selectively neutral. Mol Biol Evol 26(7):1607–1617. doi:10.1093/molbev/msp072

    Article  CAS  PubMed  Google Scholar 

  43. Mardis ER (2008) The impact of next-generation sequencing technology on genetics. Trends Genet 24(3):133–141. doi:10.1016/j.tig.2007.12.007

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work has been partly funded by a competitive funding grant from Charles Darwin University to MB as part as her PhD thesis, and by the Agence Nationale de la Recherche BDIV-07-008, project ‘Incendies Nouvelle Calédonie’. We are very grateful to the New Caledonian Gohapin tribe who has allowed us to access and sample on their land, and to Jasmin Packer, Quentin Auriac, Barbara Pianu and Viviane Degret for their help in the field. We thank Alan Andersen for providing the Leptomyrmex nigriventris specimen from the CSIRO-TERC collection.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Maïa Berman.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Berman, M., Austin, C.M. & Miller, A.D. Characterisation of the complete mitochondrial genome and 13 microsatellite loci through next-generation sequencing for the New Caledonian spider-ant Leptomyrmex pallens . Mol Biol Rep 41, 1179–1187 (2014). https://doi.org/10.1007/s11033-013-2657-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-013-2657-5

Keywords

Navigation