Advertisement

Journal of Chemical Ecology

, Volume 39, Issue 11–12, pp 1373–1384 | Cite as

Patterns of Secondary Metabolite Allocation to Fruits and Seeds in Piper reticulatum

  • S. R. WhiteheadEmail author
  • C. S. Jeffrey
  • M. D. Leonard
  • C. D. Dodson
  • L. A. Dyer
  • M. D. Bowers
Article

Abstract

Little is known about the evolution, diversity, and functional significance of secondary metabolites in reproductive plant parts, particularly fruits and seeds of plants in natural ecosystems. We compared the concentration and diversity of amides among six tissue types of Piper reticulatum: leaves, roots, flowers, unripe fruit pulp, ripe fruit pulp, and seeds. This represents the first detailed description of amides in P. reticulatum, and we identified 10 major and 3 minor compounds using GC/MS and NMR analysis. We also detected 30 additional unidentified minor amide components, many of which were restricted to one or a few plant parts. Seeds had the highest concentrations and the highest diversity of amides. Fruit pulp had intermediate concentrations and diversity that decreased with ripening. Leaves and roots had intermediate concentrations, but the lowest chemical diversity. In addition, to investigate the potential importance of amide concentration and diversity in plant defense, we measured leaf herbivory and seed damage in natural populations, and examined the relationships between amide occurrence and plant damage. We found no correlations between leaf damage and amide diversity or concentration, and no correlation between seed damage and amide concentration. The only relationship we detected was a negative correlation between seed damage and amide diversity. Together, our results provide evidence that there are strong selection pressures for fruit and seed defense independent of selection in vegetative tissues, and suggest a key role for chemical diversity in fruit-frugivore interactions.

Keywords

Amides Chemical diversity Fruit secondary metabolites Piperaceae Seed dispersal Toxic fruit 

Notes

Acknowledgments

This research was supported by National Science Foundation (NSF) grant DEB 1210884 to SRW and MDB, NSF grant DEB 0614883 to MDB and LAD, a National Geographic Waitt Grant to SRW, and an Organization for Tropical Studies (OTS) Research Fellowship to SRW. Work related to the phytochemical characterization of compounds was supported by start-up funds from the University of Nevada to CSJ and NSF grant DEB 1145609 to CSJ and LAD. We are grateful to Hannah Burk, Heather Stone, Daniel Brunelle, and Maria Obando-Quesada for assistance with sample collection and field work, and to Jason Hong for assistance with sample preparation for amide analysis. The OTS staff and especially Bernal Matarrita and Danilo Brenes provided logistical support at La Selva Biological Station, and Javier Guevara from the Ministerio del Ambiente y Energía in Costa Rica assisted with permits. Comments from two anonymous reviewers greatly improved the manuscript.

Supplementary material

10886_2013_362_MOESM1_ESM.doc (174 kb)
ESM 1 (DOC 174 kb)

References

  1. Adler LS (2000) The ecological significance of toxic nectar. Oikos 91:409–420CrossRefGoogle Scholar
  2. Adler LS, Wink M, Distl M, Lentz AJ (2006) Leaf herbivory and nutrients increase nectar alkaloids. Ecol Lett 9:960–967PubMedCrossRefGoogle Scholar
  3. Alves MN, Sartoratto A, Trigo JR (2007) Scopolamine in Brugmansia suaveolens (Solanaceae): defense, allocation, costs, and induced response. J Chem Ecol 33:297–309PubMedCrossRefGoogle Scholar
  4. Barnea A, Harborne JB, Pannell C (1993) What parts of fleshy fruits contain secondary compounds toxic to birds and why? Biochem Syst Ecol 21:421–429CrossRefGoogle Scholar
  5. Baskin JM, Baskin CC (2004) A classification system for seed dormancy. Seed Sci Res 14:1–16Google Scholar
  6. Bates D, Maechler M (2010) lme4: linear mixed-effects models using S4 classes. Version 0.999375-35. http://lme4.r-forge.r-project.org/
  7. Beckman NG (2013) The distribution of fruit and seed toxicity during development for eleven Neotropical trees and vines in Central Panama. PLoS One 8:1–19Google Scholar
  8. Berenbaum M (1985) Brementown revisited: interactions among allelochemicals in plants. In: Cooper-Driver GA, Swain T, Conn EE (eds) Recent advances in phytochemistry vol 19: chemically mediated interactions between plants and other organisms. Plenum Press, New York, pp 139–169CrossRefGoogle Scholar
  9. Berenbaum MR, Zangerl AR (1996) Phytochemical diversity: adaptation or random variation? In: Romeo JT, Saunders JA, Barbosa P (eds) Phytochemical diversity and redundancy in ecological interactions. Plenum Press, New York, pp 1–24CrossRefGoogle Scholar
  10. Bernard CB, Krishnamurty HG, Chauret D, Durst T, Philogene BJR, Sanchezvindas P, Hasbun C, Poveda L, Sanroman L, Arnason JT (1995) Insecticidal defenses of Piperaceae from the neotropics. J Chem Ecol 21:801–814PubMedCrossRefGoogle Scholar
  11. Bolker BM, Brooks ME, Clark CJ, Geange SW, Poulsen JR, Stevens MHH, White JSS (2009) Generalized linear mixed models: a practical guide for ecology and evolution. Trends Ecol Evol 24:127–135PubMedCrossRefGoogle Scholar
  12. Brown PD, Tokuhisa JG, Reichelt M, Gershenzon J (2003) Variation of glucosinolate accumulation among different organs and developmental stages of Arabidopsis thaliana. Phytochemistry 62:471–481PubMedCrossRefGoogle Scholar
  13. Burroughs LF (1970) Amino acids. In: Hulme AC (ed) The biochemistry of fruits and their products. Academic, London, pp 119–146Google Scholar
  14. Cazetta E, Schaefer HM, Galetti M (2008) Does attraction to frugivores or defense against pathogens shape fruit pulp composition? Oecologia 155:277–286PubMedCrossRefGoogle Scholar
  15. Cipollini ML (2000) Secondary metabolites of vertebrate-dispersed fruits: evidence for adaptive functions. Rev Chil Hist Nat 73:421–440CrossRefGoogle Scholar
  16. Cipollini ML, Levey DJ (1997a) Secondary metabolites of fleshy vertebrate-dispersed fruits: adaptive hypotheses and implications for seed dispersal. Am Nat 150:346–372PubMedCrossRefGoogle Scholar
  17. Cipollini ML, Levey DJ (1997b) Antifungal activity of Solanum fruit glycoalkaloids: implications for frugivory and seed dispersal. Ecology 78:799–809Google Scholar
  18. Cipollini ML, Paulk E, Mink K, Vaughn K, Fischer T (2004) Defense tradeoffs in fleshy fruits: effects of resource variation on growth, reproduction, and fruit secondary chemistry in Solanum carolinense. J Chem Ecol 30:1–17PubMedCrossRefGoogle Scholar
  19. Crozier A, Yokota T, Jaganath IB, Marks SC, Saltmarsh M, Clifford MN (2006) Secondary metabolites in fruits, vegetables, beverages and other plant based dietary components. In: Crozier A, Clifford MN, Ashihara H (eds) Plant secondary metabolites: occurrence, structure and role in the human diet. Blackwell, Oxford, pp 208–302Google Scholar
  20. De Araújo-Júnior JX, Ribeiro ÊAN, Da Silva SAS, Da Costa CDF, Alexandre-Moreira MS, Santos BVO, De O-Chaves MCO, De Medeiros IA (2011) Cardiovascular effects of two amides (piperine and piperdardine) isolated from Piper tuberculatum Jacq. Emirates J Food Agric 23:265–274Google Scholar
  21. Do Nascimento JC, Paula D, Vanderlúcia F, David JM, David JP (2012) Occurrence, biological activities and 13C NMR data of amides from Piper (Piperaceae). Quim Nova 35:2288–2311CrossRefGoogle Scholar
  22. Dyer LA, Gentry GL (2012) Caterpillars and parasitoids of a tropical lowland wet forest. Accessed on 01/07/13. http://www.caterpillars.org
  23. Dyer LA, Dodson CD, Beihoffer J, Letourneau DK (2001) Trade-offs in antiherbivore defenses in Piper cenocladum: ant mutualists versus plant secondary metabolites. J Chem Ecol 27:581–592PubMedCrossRefGoogle Scholar
  24. Dyer LA, Dodson CD, Stireman JO III, Tobler MA, Smilanich AM, Fincher RM, Letourneau DK (2003) Synergistic effects of three Piper amides on generalist and specialist herbivores. J Chem Ecol 29:2499–2514PubMedCrossRefGoogle Scholar
  25. Dyer LA, Letourneau DK, Dodson CD, Tobler MA, Stireman JO, Hsu A (2004a) Ecological causes and consequences of variation in defensive chemistry of a Neotropical shrub. Ecology 85:2795–2803CrossRefGoogle Scholar
  26. Dyer LA, Richards J, Dodson CD (2004b) Isolation, synthesis, and evolutionary ecology of Piper amides. In: Dyer LA, Palmer ADN (eds) Piper: a model genus for studies in phytochemistry, ecology, and evolution. Kluwer, New York, pp 117–139CrossRefGoogle Scholar
  27. Ehrlen J, Eriksson O (1993) Toxicity in fleshy fruits—a non-adaptive trait? Oikos 66:107–113CrossRefGoogle Scholar
  28. Eriksson O, Ehrlen J (1998) Secondary metabolites in fleshy fruits: are adaptive explanations needed? Am Nat 152:905–907PubMedCrossRefGoogle Scholar
  29. Fleming T (1991) The relationship between body size, diet, and habitat use in frugivorous bats, genus Carollia (Phyllostomidae). J Mammal 72:493–501CrossRefGoogle Scholar
  30. Fleming TH (2004) Dispersal ecology of neotropical Piper shrubs and treelets. In: Dyer LA, Palmer ADN (eds) Piper: a model genus for studies of phytochemistry, ecology, and evolution. Kluwer, New York, pp 58–77CrossRefGoogle Scholar
  31. Gershenzon J, Fontana A, Burow M, Wittstock UTE, Degenhardt J (2012) Mixtures of plant secondary metabolites: metabolic origins and ecological benefits. In: Iason GR, Dicke M, Hartley SE (eds) The ecology of plant secondary metabolites: from genes to global processes. Cambridge University Press, Cambridge, pp 56–77Google Scholar
  32. Greig N (1993) Predispersal seed predation on five Piper species in tropical rain-forest. Oecologia 93:412–420CrossRefGoogle Scholar
  33. Guo C, Yang J, Wei J, Li Y, Xu J, Jiang Y (2003) Antioxidant activities of peel, pulp and seed fractions of common fruits as determined by FRAP assay. Nutr Res 23:1719–1726CrossRefGoogle Scholar
  34. Heaney RK, Fenwick GR, Dey PM, Harborne JB (1993) Methods for glucosinolate analysis. In: Waterman PG (ed) Methods in plant biochemistry Vol. 8: alkaloids and sulfur compounds. Academic, London, pp 531–550Google Scholar
  35. Herrera CM (1982) Defense of ripe fruit from pests—its significance in relation to plant-disperser interactions. Am Nat 120:218–241CrossRefGoogle Scholar
  36. Hodisan T, Culea M, Cimpoiu C, Cot A (1998) Separation, identification and quantitative determination of free amino acids from plant extracts. J Pharm Biomed Anal 18:319–323PubMedCrossRefGoogle Scholar
  37. Hothorn T, Bretz F, Westfall P, Heiberger RM, Schuetzenmeister (2011) multcomp: simultaneous inference in general parametric models. Version 1.2–8. http://cran.r-project.org/web/packages/multcomp/index.html
  38. Hulme P (1998) Post-dispersal seed predation and seed bank persistence. Seed Sci Res 8:513–519CrossRefGoogle Scholar
  39. Iason GR, Dicke M, Hartley SE (2012) The ecology of plant secondary metabolites: from genes to global processes. Cambridge University Press, CambridgeCrossRefGoogle Scholar
  40. Irwin RE, Adler LS, Brody AK (2004) The dual role of floral traits: pollinator attraction and plant defense. Ecology 85:1503–1511CrossRefGoogle Scholar
  41. Isman MB, Duffey SS, Scudder GGE (1977) Cardenolide content of some leaf-and stem-feeding insects on temperate North American milkweeds (Asclepias spp.). Can J Zool 55:1024–1028Google Scholar
  42. Izhaki I (2002) Emodin—a secondary metabolite with multiple ecological functions in higher plants. New Phytol 155:205–217CrossRefGoogle Scholar
  43. JMP (2010) JMP Pro Version 9.0.2. SAS Institute, Inc., http://www.jmp.com/
  44. Johnson AE, Molyneux RJ, Merrill GB (1985) Chemistry of toxic range plants. Variation in pyrrolizidine alkaloid content of Senecio, Amsinckia, and Crotalaria species. J Agric Food Chem 33:50–55CrossRefGoogle Scholar
  45. Jordano P (2000) Fruits and frugivory. In: Fenner M (ed) Seeds: the ecology of regeneration in plant communities, 2nd edn. CABI Publishing, Wallingford, pp 125–166CrossRefGoogle Scholar
  46. Kessler A, Halitschke R (2009) Testing the potential for conflicting selection on floral chemical traits by pollinators and herbivores: predictions and case study. Funct Ecol 23:901–912CrossRefGoogle Scholar
  47. Kliebenstein DJ, Gershenzon J, Mitchell-Olds T (2001) Comparative quantitative trait loci mapping of aliphatic, indolic and benzylic glucosinolate production in Arabidopsis thaliana leaves and seeds. Genetics 159:359–370PubMedGoogle Scholar
  48. Laska M (1990) Olfactory discrimination ability in short-tailed fruit bat, Carollia perspicillata (Chiroptera: Phyllostomidae). J Chem Ecol 16:3291–3299PubMedCrossRefGoogle Scholar
  49. Levey DJ, Tewksbury JJ, Izhaki I, Tsahar E, Haak DC (2007) Evolutionary ecology of secondary compounds in ripe fruit: case studies with capsaicin and emodin. In: Dennis AJ, Green RJ, Schupp EW, Westcott DA (eds) Seed dispersal: theory and its application in a changing world. CABI Publishing, Oxon, pp 37–58Google Scholar
  50. Luz AIR, Zoghbi MDGB, Maia JGS (2003) The essential oils of Piper reticulatum L. and P. crassinervium HBK. Acta Amaz 33:341–343Google Scholar
  51. Maxwell A, Dabideen D, Reynolds WF, Mclean S (1998) Two 6-substituted 5,6-dihydropyran-2-ones from Piper reticulatum. J Nat Prod 61:815–816PubMedCrossRefGoogle Scholar
  52. Mccall AC, Fordyce JA (2010) Can optimal defence theory be used to predict the distribution of plant chemical defences? J Ecol 98:985–992CrossRefGoogle Scholar
  53. Mcdade LA, Bawa KS, Hespenheide HA, Hartshorn GS (1994) La Selva: ecology and natural history of a neotropical rain forest. University of Chicago Press, ChicagoGoogle Scholar
  54. Mckey D (1974) Adaptive patterns in alkaloid physiology. Am Nat 108:305–320CrossRefGoogle Scholar
  55. Mclafferty FW, Turecek F (1993) Interpretation of mass spectra, 4th edn. University Science, SausalitoGoogle Scholar
  56. Mikich SB, Bianconi GV, Maia BHLNS, Teixeira SD, Helena B, Maia NS (2003) Attraction of the fruit-eating bat Carollia perspicillata to Piper gaudichaudianum essential oil. J Chem Ecol 29:2379–2383PubMedCrossRefGoogle Scholar
  57. Minchin PR (1987) An evaluation of the relative robustness of techniques for ecological ordination. Plant Ecol 69:89–107CrossRefGoogle Scholar
  58. Moco S, Capanoglu E, Tikunov Y, Bino RJ, Boyacioglu D, Hall RD, Vervoort J, De Vos RCH (2007) Tissue specialization at the metabolite level is perceived during the development of tomato fruit. J Exp Bot 58:4131–4146PubMedCrossRefGoogle Scholar
  59. Ndakidemi PA, Dakora FD (2003) Legume seed flavonoids and nitrogenous metabolites as signals and protectants in early seedling development. Funct Plant Biol 30:729–745CrossRefGoogle Scholar
  60. Nelson CJ, Seiber JN, Brower LP (1981) Seasonal and intraplant variation of cardenolide content in the California milkweed, Asclepias eriocarpa, and implications for plant defense. J Chem Ecol 7:981–1010CrossRefGoogle Scholar
  61. Oksanen J, Blanchet FG, Kindt R, Legendre P, O’Hara RB, Simpson GL, Solymos P, Stevens MHH, Wagner H (2010) vegan: community ecology package. Version 1.17–4. http://vegan.r-forge.r-project.org/
  62. OTS (2012) Organization for Tropical Studies database: La Flora Digital de La Selva. Accessed on 12/10/12. http://sura.ots.ac.cr/local/florula3/en/index.htm
  63. Price PW, Bouton CE, Gross P, Mcpheron BA, Thompson JN, Weis AE (1980) Interactions among 3 trophic levels—influence of plants on interactions between insect herbivores and natural enemies. Annu Rev Ecol Syst 11:41–65CrossRefGoogle Scholar
  64. R-Development-Core-Team (2012) R: a language and environment for statistical computing. Version 2.15.1. http://www.R-project.org. Vienna, Austria
  65. Rhoades DF, Cates RG (1976) Toward a general theory of plant antiherbivore chemistry. Recent Adv Phytochem 10:168–213Google Scholar
  66. Richards LA, Dyer LA, Smilanich AM, Dodson CD (2010) Synergistic effects of amides from two Piper species on generalist and specialist herbivores. J Chem Ecol 36:1105–1113PubMedCrossRefGoogle Scholar
  67. Rodríguez A, Alquézar B, Peña L (2013) Fruit aromas in mature fleshy fruits as signals of readiness for predation and seed dispersal. New Phytol 197:36–48PubMedCrossRefGoogle Scholar
  68. Seigler DS, Brinker AM, Dey PM, Harborne JB (1993) Characterisation of cyanogenic glycosides, cyanolipids, nitroglycosides, organic nitro compounds and nitrile glucosides from plants. In: Waterman PG (ed) Methods in plant biochemistry vol. 8: alkaloids and sulfur compounds. Academic, London, pp 51–131Google Scholar
  69. Srinivasan K (2007) Black pepper and its pungent principle-piperine: a review of diverse physiological effects. Crit Rev Food Sci Nutr 47:735–748PubMedCrossRefGoogle Scholar
  70. Stein SE (1995) Chemical substructure identification by mass spectral library searching. J Am Soc Mass Spectrom 6:644–655PubMedGoogle Scholar
  71. Struempf H, Schondube J, Martinez Del Rio C (1999) The cyanogenic glycoside amygdalin does not deter consumption of ripe fruit by cedar waxwings. Auk 116:749–758CrossRefGoogle Scholar
  72. Suzuki T, Waller GR (1985) Purine alkaloids of the fruits of Camellia sinensis L. and Coffea arabica L. during fruit development. Ann Bot 56:537–542Google Scholar
  73. Tewksbury JJ (2002) Fruits, frugivores and the evolutionary arms race. New Phytol 156:137–139CrossRefGoogle Scholar
  74. Tewksbury JJ, Nabhan GP (2001) Directed deterrence by capsaicin in chillies. Nature 412:403–404PubMedCrossRefGoogle Scholar
  75. Tewksbury JJ, Reagan KM, Machnicki NJ, Carlo TA, Haak DC, Peñaloza ALC, Levey DJ, Penaloza ALC (2008) Evolutionary ecology of pungency in wild chilies. Proc Natl Acad Sci U S A 105:11808–11811PubMedCrossRefGoogle Scholar
  76. Thies W, Kalko EKV (2004) Phenology of Neotropical pepper plants (Piperaceae) and their association with their main dispersers, two short-tailed fruit bats, Carollia perspicillata and C. castanea (Phyllostomidae). Oikos 104:362–376CrossRefGoogle Scholar
  77. Thies W, Kalko E, Schnitzler H (1998) The roles of echolocation and olfaction in two Neotropical fruit-eating bats, Carollia perspicillata and C. castanea, feeding on Piper. Behav Ecol Sociobiol 42:397–409CrossRefGoogle Scholar
  78. Tropicos (2012) Botanical information system at the Missouri Botanical Garden. Accessed on 12/05/12. www.tropicos.org
  79. Tsahar E, Friedman J, Izhaki I (2002) Impact on fruit removal and seed predation of a secondary metabolite, emodin, in Rhamnus alaternus fruit pulp. Oikos 99:290–299CrossRefGoogle Scholar
  80. Warton DI, Hui FKC (2011) The arcsine is asinine: the analysis of proportions in ecology. Ecology 92:3–10PubMedCrossRefGoogle Scholar
  81. Waterman PG, Dey PM, Harborne JB (1993) Alkaloids: general observations. In: Waterman PG (ed) Methods in plant biochemistry vol 8: alkaloids and sulfur compounds. Academic, LondonGoogle Scholar
  82. Whitehead SR, Bowers M (2013a) Evidence for the adaptive significance of secondary compounds in vertebrate-dispersed fruits. Am Nat 182: 563–577Google Scholar
  83. Whitehead SR, Bowers MD (2013b) Iridoid and secoiridoid glycosides in a hybrid complex of bush honeysuckles (Lonicera spp., Caprifolicaceae): implications for evolutionary ecology and invasion biology. Phytochemistry 86:57–63PubMedCrossRefGoogle Scholar
  84. Whitehead SR, Poveda K (2011) Herbivore-induced changes in fruit-frugivore interactions. J Ecol 99:964–969CrossRefGoogle Scholar
  85. Wink M, Witte L (1985) Quinolizidine alkaloids in Petteria ramentacea and the infesting aphids, Aphis cytisorum. Phytochemistry 24:2567–2568CrossRefGoogle Scholar
  86. Yamaguchi LF, Freitas GC, Yoshida NC, Silva RA, Gaia AM, Silva AM, Scotti MT, Emerenciano VP, Guimarães EF, Floh EIS (2011) Chemometric analysis of ESIMS and NMR data from Piper species. J Braz Chem Soc 22:2371–2382CrossRefGoogle Scholar
  87. Zangerl AR, Rutledge CE (1996) The probability of attack and patterns of constitutive and induced defense: a test of optimal defense theory. Am Nat 147:599–608CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • S. R. Whitehead
    • 1
    Email author
  • C. S. Jeffrey
    • 2
  • M. D. Leonard
    • 2
  • C. D. Dodson
    • 3
  • L. A. Dyer
    • 4
  • M. D. Bowers
    • 1
    • 5
  1. 1.Department of Ecology and Evolutionary BiologyUniversity of Colorado at BoulderBoulderUSA
  2. 2.Department of ChemistryUniversity of Nevada at RenoRenoUSA
  3. 3.Storm Peak LaboratoryDesert Research InstituteSteamboat SpringsUSA
  4. 4.Department of BiologyUniversity of Nevada at RenoRenoUSA
  5. 5.Museum of Natural HistoryUniversity of Colorado at BoulderBoulderUSA

Personalised recommendations