Geographically separated orange and blue populations of the Amazonian poison frog Adelphobates galactonotus (Anura, Dendrobatidae) do not differ in alkaloid composition or palatability

Abstract

As is typical of chemically defended animals, poison frogs present high variability in their alkaloid-based defenses. Previous studies have shown that geographically separated color morphs of Oophaga and Dendrobates species differ in both alkaloid composition and arthropod palatability. Here, we tested the generality of that finding by studying the alkaloid composition and palatability of geographically separated blue and orange morphs of the splash-backed poison frog, Adelphobates galactonotus. We identified and quantified the alkaloid composition of each individual frog using gas chromatography–mass spectrometry and evaluated the palatability of individual secretions to arthropods conducting feeding trials with Drosophila melanogaster. Despite their conspicuous differences in color and separation on opposite sides of a large aquatic barrier, the two morphs did not differ in alkaloid composition or palatability. This result shows that both color morphs are equally chemically protected and suggests that the color variation is not driven by predator selection.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. Andriamaharavo NR, Garraffo HM, Saporito RA, Daly JW, Razafindrabe CR, Andriantsiferana M, Spande TF (2010) Roughing it: a mantellid poison frog shows greater alkaloid diversity in some disturbed habitats. J Nat Prod 73:322–330. https://doi.org/10.1021/np900721r

    CAS  Article  PubMed  Google Scholar 

  2. Bolton SK, Dickerson K, Saporito RA (2017) Variable alkaloid defense in the dendrobatid poison frog (Oophaga pumilio) are perceived as differences in palatability to arthropods. J Chem Ecol 43:273–289

    CAS  Article  Google Scholar 

  3. Brusa O, Bellati A, Meuche I, Mundy NI, Pröhl H (2013) Divergent evolution in the polymorphic granular poison-dart frog, Oophaga granulifera: genetics, coloration, advertisement calls and morphology. J Biogeogr 40:394–408. https://doi.org/10.1111/j.1365-2699.2012.02786.x

    Article  Google Scholar 

  4. Crothers L, Saporito RA, Yeager J, Lynch K, Friesen C, Richards-Zawacki CL, McGraw K, Cummings M (2016) Warning signal properties covary with toxicity but not testosterone or aggregate carotenoids in a poison frog. Evol Ecol 30:601–621. https://doi.org/10.1007/s10682-016-9830-y

    Article  Google Scholar 

  5. Daly JW, Myers CW (1967) Toxicity of Panamanian poison frogs (Dendrobates): some biological and chemical aspects. Science 156:970–973

    CAS  Article  Google Scholar 

  6. Daly JW, Secunda S, Garraffo HM, Spande TF, Wisnieski A, Cover JF Jr (1994) An uptake system for dietary alkaloids in poison frogs (Dendrobatidae). Toxicon 32:657–663

    CAS  Article  Google Scholar 

  7. Daly JW, Garraffo HM, Spande TF, Clark VC, Ma J, Ziffer H, Cover JF (2003) Evidence for an enantioselective pumiliotoxin 7-hydroxylase in dendrobatid poison frogs of the genus Dendrobates. Proc Natl Acad Sci USA 100:11092–11097. https://doi.org/10.1073/pnas.1834430100

    CAS  Article  PubMed  Google Scholar 

  8. Daly JW, Spande TF, Garraffo HM (2005) Alkaloids from amphibian skin: a tabulation of over eight-hundred compounds. J Nat Prod 68:1556–1575. https://doi.org/10.1021/np0580560

    CAS  Article  PubMed  Google Scholar 

  9. Daly JW, Garraffo HM, Spande TF, Giddings LA, Saporito RA, Vieites DR, Vences M (2008) Individual and geographic variation of skin alkaloids in three species of Madagascan poison frogs (Mantella). J Chem Ecol 34:252–279. https://doi.org/10.1007/s10886-007-9396-9

    CAS  Article  PubMed  Google Scholar 

  10. Daly JW, Ware N, Saporito RA, Spande TF, Garraffo HM (2009) N-methyldecahydroquinolines: an unexpected class of alkaloids from Amazonian poison frogs (Dendrobatidae). J Nat Prod 72:1110–1114. https://doi.org/10.1021/np900094v

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  11. Devambez I, Ali Agha M, Mitri C, Bockaert J, Parmentier ML, Marion-Poll F, Grau Y, Soustelle L (2013) Gαo is required for L-Canavanine detection in Drosophila. PLoS One. https://doi.org/10.1371/journal.pone.0063484

    Article  PubMed  PubMed Central  Google Scholar 

  12. Dyer LA, Dodson CD, Gentry G (2003) A bioassay for insect deterrent compounds found in plant and animal tissues. Phytochem Analysis 14:381–388. https://doi.org/10.1002/pca.734

    CAS  Article  Google Scholar 

  13. Fritz G, Rand AS, dePamphilis CW (1981) The aposematically colored frog, Dendrobates pumilio, is distasteful to the large, predatory ant, Paraponera clavata. Biotropica 13:158–159

    Article  Google Scholar 

  14. Frost DR (2019) Amphibian species of the world: an online reference. Version 6.0. American Museum of Natural History http://research.amnh.org/herpetology/amphibia/index.html. Accessed 12 Aug 2019

  15. Garraffo HM, Andriamaharavo NR, Vaira M, Quiroga MF, Heit C, Spande TF (2012) Alkaloids from single skins of the Argentinian toad Melanophryniscus rubriventris (Anura, Bufonidae): an unexpected variability in alkaloid profiles and a profusion of new structures. SpringerPlus 1:51. https://doi.org/10.1186/2193-1801-1-51

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  16. Grant T (2019) Outgroup sampling in phylogenetics: severity of test and successive outgroup expansion. J Zool Syst Evol Res. https://doi.org/10.1111/jzs.12317

    Article  Google Scholar 

  17. Grant T, Colombo P, Verrastro L, Saporito RA (2012) The occurrence of defensive alkaloids in non-integumentary tissues of the Brazilian red-belly toad Melanophryniscus simplex (Bufonidae). Chemoecology 22:169–178. https://doi.org/10.1007/s00049-012-0107-9

    CAS  Article  Google Scholar 

  18. Grant T, Rada M, Anganoy-Criollo M, Batista A, Dias PH, Jeckel AM, Machado DJ, Rueda-Almonacid JV (2017) Phylogenetic systematics of Dart-Poison frogs and their relatives revisited (Anura: Dendrobatoidea). S Am J Herpetol 12:S1–S90. https://doi.org/10.2994/SAJH-D-17-00017.1

    Article  Google Scholar 

  19. Gray HM, Kaiser H, Green DM (2010) Does alkaloid sequestration protect the green poison frog, Dendrobates auratus, from predator attacks? Salamandra 46:235–238

    Google Scholar 

  20. Hantak MM, Grant T, Reinsch S, Mcginnity D, Loring M, Toyooka N, Saporito RA (2013) Dietary alkaloid sequestration in a poison frog: an experimental test of alkaloid uptake in Melanophryniscus stelzneri (Bufonidae). J Chem Ecol 39:1400–1406. https://doi.org/10.1007/s10886-013-0361-5

    CAS  Article  PubMed  Google Scholar 

  21. Hegna RH, Saporito RA, Gerow KG, Donnelly MA (2011) Contrasting colors of an aposematic poison frog do not affect predation. Ann Zool Fenn 48:29–38

    Article  Google Scholar 

  22. Hoogmoed MS, Ávila-Pires TCS (2012) Inventory of color polymorphism in populations of Dendrobates galactonotus (Anura: Dendrobatidae), a poison frog endemic to Brazil. Phyllomedusa 11:95–115

    Article  Google Scholar 

  23. Jeckel AM, Grant T, Saporito RA (2015a) Sequestered and synthesized chemical defenses in the poison frog Melanophryniscus moreirae. J Chem Ecol 41:505–512. https://doi.org/10.1007/s10886-015-0578-6

    CAS  Article  PubMed  Google Scholar 

  24. Jeckel AM, Saporito RA, Grant T (2015b) The relationship between poison frog chemical defenses and age, body size, and sex. Front Zool 12:27. https://doi.org/10.1186/s12983-015-0120-2

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  25. Lawrence JP, Rojas B, Fouquet A, Mappes J, Blanchette A, Saporito RA, Bosque RJ, Courtois EA, Noonan BP (2019) Weak warning signals can persist in the absence of gene flow. Proc Nat Acad Sci USA. https://doi.org/10.1073/pnas.1901872116

    Article  PubMed  Google Scholar 

  26. Lee Y, Moon SJ, Wang Y, Montell C (2015) A drosophila gustatory receptor required for strychnine sensation. Chem Senses 40:525–533. https://doi.org/10.1093/chemse/bjv038

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  27. Lillywhite HB, Shine R, Jacobson E, Denardo DF, Gordon MS, Navas CA, Wang T, Seymour RS, Storey KB, Heatwole H, Heard D, Brattstrom B, Burghardt GM (2017) Anesthesia and euthanasia of amphibians and reptiles used in scientific research: should hypothermia and freezing be prohibited? Bioscience 67:53–61. https://doi.org/10.1093/biosci/biw143

    Article  Google Scholar 

  28. Maan ME, Cummings ME (2008) Female preferences for aposematic signal components in a polymorphic poison frog. Evolution 62:2334–2345. https://doi.org/10.1111/j.1558-5646.2008.00454.x

    Article  PubMed  Google Scholar 

  29. Maan ME, Cummings ME (2012) Poison frog colors are honest signals of toxicity, particularly for bird predators. Am Nat 179:1–14. https://doi.org/10.1086/663197

    Article  Google Scholar 

  30. McGugan JR, Byrd GD, Roland AB, Caty SN, Kabir N, Tapia EE, Trauger SA, Coloma LA, Connell LAO (2016) Ant and mite diversity drives toxin variation in the little devil poison frog. J Chem Ecol 42:537–551. https://doi.org/10.1007/s10886-016-0715-x

    CAS  Article  PubMed  Google Scholar 

  31. Meunier N, Marion-Poll F, Rospars JP, Tanimura T (2003) Peripheral coding of bitter taste in Drosophila. J Neurobiol 56:139–152. https://doi.org/10.1002/neu.10235

    Article  PubMed  Google Scholar 

  32. Murray EM, Bolton SK, Berg T, Saporito RA (2016) Arthropod predation in a dendrobatid poison frog: does frog life stage matter? Zoology 119:169–174

    Article  Google Scholar 

  33. Myers CW, Daly JW (1976) Preliminary evaluation of skin toxins and vocalizations in taxonomic and evolutionary studies of poison-dart frogs (Dendrobatidae). B Am Mus Nat Hist 157:173–262

    Google Scholar 

  34. Noonan BP, Comeault AA (2008) The role of predator selection on polymorphic aposematic poison frogs. Biol Lett 5:51–54. https://doi.org/10.1098/rsbl.2008.0586

    Article  PubMed Central  Google Scholar 

  35. Patrick LD, Sasa M (2009) Phenotypic and molecular variation in the green and black poison-dart frog Dendrobates auratus (Anura: Dendrobatidae) from Costa Rica. Rev Biol Trop 57:313–321

    Google Scholar 

  36. Rojas D, Stow A, Amézquita A, Simões PI, Lima AP (2015) No predatory bias with respect to colour familiarity for the aposematic Adelphobates galactonotus (Anura: Dendrobatidae). Behaviour 152:1–21. https://doi.org/10.1163/1568539X-00003297

    Article  Google Scholar 

  37. R Core Team (2019) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. https://www.R-project.org/

  38. Santos JC, Cannatella DC (2011) Phenotypic integration emerges from aposematism and scale in poison frogs. Proc Natl Acad Sci USA 108:6175–6180. https://doi.org/10.1073/pnas.1010952108

    Article  PubMed  Google Scholar 

  39. Saporito RA, Donnelly MA, Garraffo HM, Spande TF, Daly JW (2006) Geographic and seasonal variation in alkaloid-based chemical defenses of Dendrobates pumilio from Bocas del Toro, Panama. J Chem Ecol 32:795–814. https://doi.org/10.1007/s10886-006-9034-y

    CAS  Article  PubMed  Google Scholar 

  40. Saporito RA, Donnelly MA, Jain P, Garraffo HM, Spande TF, Daly JW (2007a) Spatial and temporal patterns of alkaloid variation in the poison frog Oophaga pumilio in Costa Rica and Panama over 30 years. Toxicon 50:757–778. https://doi.org/10.1016/j.toxicon.2007.06.022

    CAS  Article  PubMed  Google Scholar 

  41. Saporito RA, Zuercher R, Roberts M, Gerow KG, Donnelly MA (2007b) Experimental evidence for aposematism in the dendrobatid poison frog Oophaga pumilio. Copeia 2007:1006–1011

    Article  Google Scholar 

  42. Saporito RA, Spande TF, Garraffo HM, Donnelly MA (2009) Arthropod alkaloids in poison frogs: a review of the ‘Dietary Hypothesis’. Heterocycles 79:277–297. https://doi.org/10.3987/REV-08-SR(D)11

    CAS  Article  Google Scholar 

  43. Saporito RA, Donnelly MA, Madden AA, Garraffo HM, Spande TF (2010a) Sex-related differences in alkaloid chemical defenses of the dendrobatid frog Oophaga pumilio from Cayo Nancy, Bocas del Toro, Panama. J Nat Prod 73:317–321. https://doi.org/10.1021/np900702d

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  44. Saporito RA, Isola M, Maccachero VC, Condon K, Donnelly MA (2010b) Ontogenetic scaling of poison glands in a dendrobatid poison frog. J Zool 282:238–245. https://doi.org/10.1111/j.1469-7998.2010.00732.x

    Article  Google Scholar 

  45. Saporito RA, Donnelly MA, Spande TF, Garraffo HM (2012) A review of chemical ecology in poison frogs. Chemoecology 22:159–168. https://doi.org/10.1007/s00049-011-0088-0

    CAS  Article  Google Scholar 

  46. Saporito RA, Grant T (2018) Comment on Amézquita et al. (2017) “Conspicuousness, color resemblance, and toxicity in geographically diverging mimicry: the pan-Amazonian frog Allobates femoralis”. Evolution 72:1009–1014. https://doi.org/10.1111/evo.13468

    CAS  Article  PubMed  Google Scholar 

  47. Schulte LM, Saporito RA, Davison I, Summers K (2017) The palatability of Neotropical poison frogs in predator-prey systems: do alkaloids make the difference? Biotropica 49:23–26. https://doi.org/10.1111/btp.12404

    Article  Google Scholar 

  48. Sellier MJ, Reeb P, Marion-Poll F (2011) Consumption of bitter alkaloids in Drosophila melanogaster in multiple-choice test conditions. Chem Senses 36:323–334. https://doi.org/10.1093/chemse/bjq133

    CAS  Article  PubMed  Google Scholar 

  49. Silverstone PA (1975) A revision of the poison-arrow frogs of the genus Dendrobates Wagler. Nat Hist Mus L A Sci Bull 21:1–55

    Google Scholar 

  50. Speed MP, Ruxton GD, Mappes J, Sherratt TN (2012) Why are defensive toxins so variable? An evolutionary perspective. Biol Rev 87:874–884. https://doi.org/10.1111/j.1469-185X.2012.00228.x

    Article  PubMed  Google Scholar 

  51. Stuckert AM, Saporito RA, Venegas PJ, Summers K (2014) Alkaloid defenses of co-mimics in a putative Müllerian mimetic radiation. BMC Evol Biol 14:1–8. https://doi.org/10.1186/1471-2148-14-76

    CAS  Article  Google Scholar 

  52. Stuckert AM, Saporito RA, Summers K (2018) An empirical test indicates only qualitatively honest aposematic signaling within a population of vertebrates. J Herpetol 52:201–208. https://doi.org/10.1670/17-047

    Article  Google Scholar 

  53. Stynoski JL, Shelton G, Stynoski P (2014a) Maternally derived chemical defences are an effective deterrent against some predators of poison frog tadpoles (Oophaga pumilio). Biol Lett 10:20140187. https://doi.org/10.1098/rsbl.2014.0187

    Article  PubMed  PubMed Central  Google Scholar 

  54. Stynoski J, Torres-Mendoza Y, Sasa-Marin M, Saporito RA (2014b) Evidence of maternal provisioning of alkaloid-based chemical defenses in the strawberry poison frog Oophaga pumilio. Ecology 95:587–593

    Article  Google Scholar 

  55. Szelistowski WA (1985) Unpalatability of the poison arrow frog Dendrobates pumilio to the ctenid spider Cupiennius coccineus. Biotropica 17:345–346

    Article  Google Scholar 

  56. Wang IJ (2011) Inversely related aposematic traits: reduced conspicuousness evolves with increased toxicity in a polymorphic poison-dart frog. Evolution 65:1637–1649. https://doi.org/10.1111/j.1558-5646.2011.01257.x

    Article  PubMed  Google Scholar 

  57. Yang Y, Servedio MR, Richards-Zawacki CL (2019) Imprinting sets the stage for speciation. Nature 574:99–102. https://doi.org/10.1038/s41586-019-1599-z

    CAS  Article  PubMed  Google Scholar 

Download references

Acknowledgements

We thank M. Rada, J.J. Ospina-Sarria, C.A. Lopes, S. Andrade, the São Sebastião and Vila do Bravo communities, and the crew of Estação Científica Ferreira Penna (Museu Paraense Emílio Goeldi, Universidade Federal do Pará) for assistance during fieldwork. Specimen collection (license numbers 13173-2 and 54640-1) and export (authorization number 17BR025049/DF) permits were issued by Instituto Chico Mendes de Conservação da Biodiversidade (ICMBIO)/Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis (IBAMA). This study was approved by the Ethics Committee on Animal Use of the Institute of Biosciences, University of São Paulo (CEUA Protocol 268/2016). We thank M. Nichols for his assistance in maintaining the GC–MS and Kresge Foundation and Colleran-Weaver for funding. This research was supported by the Brazilian Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq Proc. 306823/2017-9) and Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP Proc. 2012/10000-5, 2016/09999-9, 2018/15425-0).

Author information

Affiliations

Authors

Contributions

AMJ, RAS, and TG contributed to the study conception and design. Specimens were collected by AMJ and TG. Chemical analysis was performed by AMJ and RAS. Palatability tests were performed by SK and RAS. The first draft of the manuscript was written by AMJ and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Taran Grant.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Handling Editor: Marko Rohlfs.

Electronic supplementary material

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Jeckel, A.M., Kocheff, S., Saporito, R.A. et al. Geographically separated orange and blue populations of the Amazonian poison frog Adelphobates galactonotus (Anura, Dendrobatidae) do not differ in alkaloid composition or palatability. Chemoecology 29, 225–234 (2019). https://doi.org/10.1007/s00049-019-00291-3

Download citation

Keywords

  • Aposematism
  • Chemical defense
  • GC–MS
  • Polychromatism
  • Polytypism