Chromosomal rearrangements maintain a polymorphic supergene controlling butterfly mimicry

Journal name:
Nature
Volume:
477,
Pages:
203–206
Date published:
DOI:
doi:10.1038/nature10341
Received
Accepted
Published online

Supergenes are tight clusters of loci that facilitate the co-segregation of adaptive variation, providing integrated control of complex adaptive phenotypes1. Polymorphic supergenes, in which specific combinations of traits are maintained within a single population, were first described for ‘pin’ and ‘thrum’ floral types in Primula1 and Fagopyrum2, but classic examples are also found in insect mimicry3, 4, 5 and snail morphology6. Understanding the evolutionary mechanisms that generate these co-adapted gene sets, as well as the mode of limiting the production of unfit recombinant forms, remains a substantial challenge7, 8, 9, 10. Here we show that individual wing-pattern morphs in the polymorphic mimetic butterfly Heliconius numata are associated with different genomic rearrangements at the supergene locus P. These rearrangements tighten the genetic linkage between at least two colour-pattern loci that are known to recombine in closely related species9, 10, 11, with complete suppression of recombination being observed in experimental crosses across a 400-kilobase interval containing at least 18 genes. In natural populations, notable patterns of linkage disequilibrium (LD) are observed across the entire P region. The resulting divergent haplotype clades and inversion breakpoints are found in complete association with wing-pattern morphs. Our results indicate that allelic combinations at known wing-patterning loci have become locked together in a polymorphic rearrangement at the P locus, forming a supergene that acts as a simple switch between complex adaptive phenotypes found in sympatry. These findings highlight how genomic rearrangements can have a central role in the coexistence of adaptive phenotypes involving several genes acting in concert, by locally limiting recombination and gene flow.

At a glance

Figures

  1. Supergene alleles and mimicry polymorphism in H. numata.
    Figure 1: Supergene alleles and mimicry polymorphism in H. numata.

    Polymorphic forms of H.numata each mimic different models in the distantly related genus Melinaea (Nymphalidae: Danainae). Each form is controlled by a specific allele of the supergene P, with increasing dominance shown from left to right4, 9. Two parapatric regions of northeastern Peru (T, Tarapoto and Andean valleys; Y, Yurimaguas and Amazon lowlands) harbour different mimicry assemblages15; dominance (<) is nearly complete between forms within each region, but is incomplete (~) between certain pairs of alleles from parapatric regions. In all other species studied in the genus Heliconius, wing pattern is controlled by several large-effect loci on different chromosomes. In H.melpomene the HmYbHmSbHmN complex is situated in the orthologous position to the H.numata P supergene9. LG, linkage group.

  2. Fine-scale mapping and nucleotide variation at the P supergene in H.[thinsp]numata.
    Figure 2: Fine-scale mapping and nucleotide variation at the P supergene in H.numata.

    a, Fine-scale linkage mapping of the supergene P (indicated by red arrows) to the interval bounded by genes HN00020 and HN00041. Recombinants were observed in crosses totalling 366 individuals. Blue arrows indicate the position of the recombining loci HmYb and HmSb in H.melpomene11. HmN, which is known to be part of this cluster of loci, is not fine-mapped in H.melpomene9, 11. Coloured blocks represent annotated gene regions on forward and reverse strands (see Supplementary Fig. 1 and Supplementary Table 3 for details). b, Association of SNP variation with mimicry polymorphism in a sample of 25 silvana and 34 aurora/arcuella individuals from a single population. Markers genotyped across rearrangements BP1 (pink) and BP2 (blue) show perfect association of SNP variation with wing pattern. No association was found in the flanking region from markers Fox (Hn00106) to BmSuc (Hn00019), or at 12 unlinked loci (green). The association decays more slowly in the direction of loci Bm5536 (GCP), Bm5586 (NudC) and Bm5593 (Srp68). c, LD heat map. Perfect LD (genotypic correlation coefficient r2 = 1) is found across 580kb spanning the BP1 and BP2 rearrangements (n = 59). LD decays rapidly outside this interval, although strong within-marker LD remains at HN00021. Markers that are unlinked to P show little LD with each other or with P. d, Haplotype network for marker LRR (Hn00024) in the Yurimaguas population, coloured according to wing-pattern phenotype. Haplotype clades separated by seven fixed differences are in complete association with wing pattern, taking into account dominance relationships. Similar haplotype clades were found for all loci genotyped within P, and across the Amazon basin, but not for genes flanking P or in unlinked regions (Supplementary Figs 4 and 6).

  3. Chromosomal rearrangements associated with the supergene in natural populations.
    Figure 3: Chromosomal rearrangements associated with the supergene in natural populations.

    a, Comparison of the gene orders found in the H.numata BAC library and wild populations. The rearrangements involve the 400-kb segment from genes HM00023 to HM00040 (ERCC6) (BP1, clones 24I10 and 45B17), and the adjacent 180-kb segment from gene HN00041 (penguin) and HN00053 (lethal (2) giant larvae homologue) (BP2, clone 38G4). Genes closest to the breakpoints are shown. b, Long-range PCR assays across alternative breakpoints (BP0, BP1 and BP2) in wild populations show a perfect association of the polymorphic gene orders with mimicry variation in four morphs from natural populations of Eastern Peru15, following the dominance relationships (Fig. 1). The Yurimaguas population segregates primarily for silvana and aurora/arcuella forms, associated with BP0 and BP2, respectively. The Tarapoto population segregates mainly for tarapotensis and bicoloratus forms, associated with BP2 and BP1, respectively. This population also harbours recessive illustris alleles associated with BP0 (Supplementary Table 6).

References

  1. Mather, K. The genetical architecture of heterostyly in Primula sinensis. Evolution 4, 340352 (1950)
  2. Garber, R. J. & Quisenberry, K. S. The inheritance of length of style in buckwheat. J. Agric. Res. 34, 181183 (1927)
  3. Clarke, C. A., Sheppard, P. M. & Thornton, I. W. B. The genetics of the mimetic butterfly Papilio memnon. Philos. Trans. R. Soc. Lond. B 254, 3789 (1968)
  4. Brown, K. S. & Benson, W. W. Adaptive polymorphism associated with multiple Müllerian mimicry in Heliconius numata. Biotropica 6, 205228 (1974)
  5. Nijhout, H. F. Polymorphic mimicry in Papilio dardanus: mosaic dominance, big effects, and origins. Evol. Dev. 5, 579592 (2003)
  6. Murray, J. & Clarke, B. Supergenes in polymorphic land snails—examples from genus Partula. Genetics 74, S188S189 (1973)
  7. Kirkpatrick, M. & Barton, N. Chromosome inversions, local adaptation and speciation. Genetics 173, 419434 (2006)
  8. Manfield, I. W. et al. Molecular characterization of DNA sequences from the Primula vulgaris S-locus. J. Exp. Bot. 56, 11771188 (2005)
  9. Joron, M. et al. A conserved supergene locus controls colour pattern diversity in Heliconius butterflies. PLoS Biol. 4, e303 (2006)
  10. Baxter, S. W. et al. Genomic hotspots for adaptation: the population genetics of Müllerian mimicry in the Heliconius melpomene clade. PLoS Genet. 6, e1000794 (2010)
  11. Ferguson, L. et al. Characterization of a hotspot for mimicry: Assembly of a butterfly wing transcriptome to genomic sequence at the HmYb/Sb locus. Mol. Ecol. 19, 240254 (2010)
  12. Hoffmann, A. A. & Rieseberg, L. H. Revisiting the impact of inversions in evolution: from population genetic markers to drivers of adaptive shifts and speciation? Annu. Rev. Ecol. Evol. Syst. 39, 2142 (2008)
  13. Pinho, C. & Hey, J. Divergence with gene flow: Models and data. Annu. Rev. Ecol. Evol. Syst. 41, 215230 (2010)
  14. Clark, R. et al. Colour pattern specification in the Mocker swallowtail Papilio dardanus: the transcription factor invected is a candidate for the mimicry locus H. Proc. R. Soc. B 275, 11811188 (2008)
  15. Joron, M., Wynne, I. R., Lamas, G. & Mallet, J. Variable selection and the coexistence of multiple mimetic forms of the butterfly Heliconius numata. Evol. Ecol. 13, 721754 (1999)
  16. Turner, J. R. G. in The biology of butterflies Vol. 11 (eds Vane-Wright, R. I. & Ackery, P. R.) 141161 (Academic, 1984)
  17. Charlesworth, D. & Charlesworth, B. Theoretical genetics of Batesian mimicry. II. Evolution of supergenes. J. Theor. Biol. 55, 305324 (1975)
  18. Alvarez, G. & Zapata, C. Conditions for protected inversion polymorphism under supergene selection. Genetics 146, 717722 (1997)
  19. Hatadani, L. M., Baptista, J. C. R., Souza, W. N. & Klaczko, L. B. Colour polymorphism in Drosophila mediopunctata: genetic (chromosomal) analysis and nonrandom association with chromosome inversions. Heredity 93, 525534 (2004)
  20. Lowry, D. B. & Willis, J. H. A widespread chromosomal inversion polymorphism contributes to a major life-history transition, local adaptation, and reproductive isolation. PLoS Biol. 8, e1000500 (2010)
  21. Jones, R. T., Salazar, P., ffrench-Constant, R. H., Jiggins, C. D. & Joron, M. Evolution of a mimicry supergene from a multilocus architecture. Proc. R. Soc. B doi:10.1098/rspb.2011.0882 (2011)
  22. Papa, R. et al. Highly conserved gene order and numerous novel repetitive elements in genomic regions linked to wing pattern variation in Heliconius butterflies. BMC Genomics 9, 345 (2008)
  23. Pringle, E. G. et al. Synteny and chromosome evolution in the lepidoptera: Evidence from mapping in Heliconius melpomene. Genetics 177, 417426 (2007)
  24. Counterman, B. A. et al. Genomic hotspots for adaptation: the population genetics of Müllerian mimicry in Heliconius erato. PLoS Genet. 6, e1000796 (2010)
  25. Dobzhansky, T. Genetics of natural populations. XIV. A response of certain gene arrangements in the third chromosome of Drosophila pseudoobscura to natural selection. Genetics 32, 142160 (1947)
  26. Saenko, S. V., Brakefield, P. M. & Beldade, P. Single locus affects embryonic segment polarity and multiple aspects of an adult evolutionary novelty. BMC Biol. 8, 111 (2010)
  27. van't Hof, A. E., Edmonds, N., Dalíková, M., Marec, F. & Saccheri, I. J. Industrial melanism in British peppered moths has a singular and recent mutational origin. Science 332, 958960 (2011)
  28. Kronforst, M. R. et al. Linkage of butterfly mate preference and wing color preference cue at the genomic location of wingless. Proc. Natl Acad. Sci. USA 103, 65756580 (2006)
  29. Chamberlain, N. L., Hill, R. I., Kapan, D. D., Gilbert, L. E. & Kronforst, M. R. Polymorphic butterfly reveals the missing link in ecological speciation. Science 326, 847850 (2009)
  30. Merrill, R. M., Van Schooten, B., Scott, J. A. & Jiggins, C. D. Pervasive genetic associations between traits causing reproductive isolation in Heliconius butterflies. Proc. R. Soc. B 278, 511518 (2010)
  31. Frazer, K. A., Pachter, L., Poliakov, A., Rubin, E. M. & Dubchak, I. VISTA: computational tools for comparative genomics. Nucleic Acids Res. 32, W273W279 (2004)
  32. Mayor, C. et al. VISTA: visualizing global DNA sequence alignments of arbitrary length. Bioinformatics 16, 1046 (2000)
  33. Cantarel, B. L. et al. MAKER: An easy-to-use annotation pipeline designed for emerging model organism genomes. Genome Res. 18, 188196 (2008)
  34. Chevreux, B. et al. Using the miraEST assembler for reliable and automated mRNA transcript assembly and SNP detection in sequenced ESTs. Genome Res. 14, 11471159 (2004)
  35. Salazar, C., Jiggins, C. D., Taylor, J. E., Kronforst, M. R. & Linares, M. Gene flow and the genealogical history of Heliconius heurippa. BMC Evol. Biol. 8, 132 (2008)
  36. Brown, K. S. An illustrated key to the silvaniform Heliconius (Lepidoptera: Nymphalidae) with descriptions of new subspecies. Trans. Am. Entomol. Soc. 102, 373484 (1976)
  37. Zaykin, D. V., Pudovkin, A. & Weir, B. S. Correlation-based inference for linkage disequilibrium with multiple alleles. Genetics 180, 533545 (2008)
  38. Stephens, M. & Donnelly, P. A comparison of Bayesian methods for haplotype reconstruction from population genotype data. Am. J. Hum. Genet. 73, 11621169 (2003)
  39. Polzin, T. & Daneshmand, S. V. On Steiner trees and minimum spanning trees in hypergraphs. Oper. Res. Lett. 31, 1220 (2003)
  40. Forster, P., Torroni, A., Renfrew, C. & Rohl, A. Phylogenetic star contraction applied to Asian and Papuan mtDNA evolution. Mol. Biol. Evol. 18, 18641881 (2001)
  41. Librado, P. & Rozas, J. DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25, 14511452 (2009)
  42. Hudson, R. R., Slatkin, M. & Maddison, W. P. Estimation of levels of gene flow from DNA-sequence data. Genetics 132, 583589 (1992)

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Author information

  1. These authors contributed equally to this work.

    • Lise Frezal &
    • Robert T. Jones

Affiliations

  1. CNRS UMR 7205, Muséum National d’Histoire Naturelle, CP50, 45 Rue Buffon, 75005 Paris, France

    • Mathieu Joron,
    • Lise Frezal &
    • Annabel Whibley
  2. Institute of Evolutionary Biology, University of Edinburgh, Ashworth Laboratories, King’s Buildings, West Mains Road, Edinburgh EH9 3JT, UK

    • Mathieu Joron &
    • Michel Becuwe
  3. Institute of Biology, Leiden University, Postbus 9505, 2300 RA Leiden, The Netherlands

    • Mathieu Joron
  4. Centre for Ecology and Conservation, School of Biosciences, University of Exeter, Cornwall Campus, Penryn, Cornwall TR10 9EZ, UK

    • Robert T. Jones,
    • Nicola L. Chamberlain,
    • Paul A. Wilkinson &
    • Richard H. ffrench-Constant
  5. Department of Genetics, Bio21 Institute, University of Melbourne, 30 Flemington Road, Parkville, 3010 Victoria, Australia

    • Siu F. Lee
  6. Department of Biology, Ecology and Evolution, University of Fribourg, Chemin du Musée 10, CH-1700 Fribourg, Switzerland

    • Christoph R. Haag
  7. Department of Zoology, University of Cambridge, Downing Street, Cambridge CB2 3EJ, UK

    • Simon W. Baxter,
    • Laura Ferguson &
    • Chris D. Jiggins
  8. Smithsonian Tropical Research Institute, NAOS island, Causeway Amador, Panamá, República de Panamá

    • Camilo Salazar
  9. The Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1HH, UK

    • Claire Davidson,
    • Richard Clark,
    • Michael A. Quail,
    • Helen Beasley,
    • Rebecca Glithero,
    • Christine Lloyd,
    • Sarah Sims,
    • Matthew C. Jones &
    • Jane Rogers

Contributions

M.J., C.D.J. and R.H.ff.-C. designed the study and contributed to all stages of the project. M.J., L. Frezal and R.T.J. performed the principal experiments and data analysis, with assistance from N.L.C., S.W.B., S.F.L., M.B., C.S., L. Ferguson, C.R.H., A.W. and P.A.W. BAC clone sequencing was carried out by C.D., R.G., C.L., R.C., H.B., S.S., J.R., M.C.J. and M.A.Q. M.J., A.W., C.D.J. and R.H.ff.-C. co-wrote the manuscript with input from all authors.

Competing financial interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to:

GenBank accessions for BAC clone sequences: FP885863, FP476061, FP565803, FP476023, CU856181, FP885878, FP476047, FP885857, CU856182, CU655868, FP885879, FP885861, FP885880, FP885855, CU914733, FP475989, CU655869, CU914734, CU633161, CU638865, CU856175, FP884220 and FP236755. Accessions for 1364 marker sequences: JN173798–JN175161.

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Supplementary information

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  1. Supplementary Information (2.6M)

    The file contains Supplementary Figures 1-6 with legends, Supplementary Tables 1-9 and additional references.

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