, Volume 25, Issue 6, pp 293–302 | Cite as

Habitat-specific divergence of procyanidins in Protium subserratum (Burseraceae)

  • John Lokvam
  • Margaret R. Metz
  • Gary R. Takeoka
  • Lien Nguyen
  • Paul V. A. Fine
Original Article


In Amazonian Peru, the neotropical tree Protium subserratum Engl. (Burseraceae) occurs as distinct ecotypes on low nutrient white-sand (WS), intermediate fertility brown-sand (BS), and nutrient-rich clay (CS) soils. Genetic analysis indicates that these ecotypes are undergoing incipient speciation. Possible drivers of this divergence are habitat-specific herbivore faunas and differing resource availabilities. Protium subserratum, therefore, provides an ideal opportunity to investigate how defense chemistry evolves during lineage divergence. WS and BS races of P. subserratum are host to largely non-overlapping herbivore communities and they differ in chlorogenic acid, flavonoid, and oxidized terpene chemistry. Here, we investigate how another important class of anti-herbivore chemicals, procyanidins (PCs), varies among the ecotypes. We isolated the PCs from leaves of juvenile and adult trees from each ecotype and used spectroscopic and chemical techniques to characterize the chemical structures of their component monomers. We found that WS, BS, and CS ecotypes accumulate ca. 17 % of leaf dry weight as PCs. Within ecotypes, we found very little difference in PC type, neither by site nor by life stage. Among ecotypes, however, we observed a marked divergence in PC composition that arose at least in part from differences in their terminal and extension subunits. In addition, the average polymer length of BS and CS PCs was significantly greater than in WS ecotypes. We conclude that phenotypic differences in PCs in the WS versus BS and CS ecotypes of P. subserratum are consistent with selection by herbivores in different soil types that differ strongly in nutrient availability and may contribute to the evolution of habitat specialization.


Protium subserratum Procyanidin Defense chemistry Herbivory Habitat specialization Speciation 



We thank the Ministerio del Ambiente of Peru for providing research and export permits. We thank Carlos Rivera of SERNANP-Allpahuayo-Mishana and the Instituto de Investigaciones de la Amazonia Peruana (IIAP) for institutional and logistical support. We would like to thank Italo Mesones, Magno Vásquez Pilco, J. Milagros Ayarza Zuñiga, Julio Sanchez for field assistance, and Leslie Harden for high-resolution mass spectrometry. We also thank Diego Salazar Amoretti for helpful suggestions to the manuscript. Funding was provided by NSF DEB 1254214 to PVAF.

Conflict of interest

The authors declare they have no conflict of interest.


  1. Barbehenn RV, Constabel CP (2011) Tannins in plant-herbivore interactions. Phytochemistry 72:1551–1565. doi: 10.1016/j.phytochem.2011.01.040 CrossRefPubMedGoogle Scholar
  2. Barbehenn RV, Jones CP, Karonen M, Salminen JP (2006) Tannin composition affects the oxidative activities of tree leaves. J Chem Ecol 32:2235–2251. doi: 10.1007/s10886-006-9142-8 CrossRefPubMedGoogle Scholar
  3. Barone JA (2000) Comparison of herbivores and herbivory in the canopy and understory for two tropical tree species. Biotropica 32:307–317. doi: 10.1111/j.1744-7429.2000.tb00474.x CrossRefGoogle Scholar
  4. Barton KE, Koricheva J (2010) The ontogeny of plant defense and herbivory: characterizing general patterns using meta-analysis. Am Nat 175:481–493. doi: 10.1086/650722 CrossRefPubMedGoogle Scholar
  5. Basset Y, Charles EC, Novotny V (1999) Insect herbivores on parent trees and conspecific seedlings in a rain forest in Guyana. Selbyana 20:146–158Google Scholar
  6. Bates D, Maechler M, Bolker B, Walker S (2014) Ime4: Linear mixed-effects models using Eigen and S4.
  7. Boege K, Marquis RJ (2005) Facing herbivory as you grow up: the ontogeny of resistance in plants. Trends Ecol Evol 20:441–448. doi: 10.1016/j.tree.2005.05.001 CrossRefPubMedGoogle Scholar
  8. Chougule NP, Bonning BC (2012) Toxins for transgenic resistance to hemipteran pests. Toxins 4:405–429. doi: 10.3390/toxins4060405 PubMedCentralCrossRefPubMedGoogle Scholar
  9. Coley PD, Barone JA (1996) Herbivory and plant defenses in tropical forests. Annu Rev Ecol Syst 27:305–335CrossRefGoogle Scholar
  10. Coley PD, Bryant JP, Chapin FS (1985) Resource availability and plant anti-herbivore defense. Science 230:895–899CrossRefPubMedGoogle Scholar
  11. Dow JAT (1992) pH gradients in lepidopteran midgut. J Exp Biol 172:355–375PubMedGoogle Scholar
  12. Feeny PR (1969) Inhibitory effect of oak leaf tannins on the hydrolysis of proteins by trypsin. Phytochemistry 8:2119–2126CrossRefGoogle Scholar
  13. Fine PVA, Miller ZJ, Mesones I, Irazuzta S, Appel HM, Stevens MHH, Saaksjarvi I, Schultz LC, Coley PD (2006) The growth-defense trade-off and habitat specialization by plants in Amazonian forests. Ecology 87:S150–S162CrossRefPubMedGoogle Scholar
  14. Fine PVA, Garcia-Villacorta R, Pitman N, Mesones I, Kembrel SW (2010) Herbivores promote habitat specialization by trees in Amazonian forests. Ann MS Bot G 97:283–305CrossRefGoogle Scholar
  15. Fine PVA, Metz MR, Lokvam J, Mesones I, Milagros ZAJ, Lamarre GPA, Vásquez PM, Baraloto C (2013a) Insect herbivory, chemical innovation and the evolution of habitat specialization in Amazonian trees. Ecology 94:1764–1765CrossRefPubMedGoogle Scholar
  16. Fine PVA, Zapata F, Daly DC, Mesones I, Misiewicz TM, Cooper HF, Barbosa CEA (2013b) The importance of environmental heterogeneity and spatial distance in generating phylogeographic structure in edaphic specialist and generalist tree species of Protium (Burseraceae) across the Amazon Basin. J Biogeogr 40:646–661. doi: 10.1111/j.1365-2699.2011.02645.x CrossRefGoogle Scholar
  17. Gentry AH (1986) Endemism in tropical versus temperate plant communities. In: Soulé ME (ed) Conservation biology: the science of scarcity and diversity. Sinauer, Sunderland, MA, pp 153–181Google Scholar
  18. Goodger JQD, Choo TYS, Woodrow IE (2007) Ontogenetic and temporal trajectories of chemical defence in a cyanogenic eucalypt. Oecologia 153:799–808. doi: 10.1007/s00442-007-0787-y CrossRefPubMedGoogle Scholar
  19. Hagerman AE, Butler LG (1978) Protein precipitation method for the quantitative determination of tannins. J Agric Food Chem 26:809–812CrossRefGoogle Scholar
  20. Herms DA, Mattson WJ (1992) The dilemma of plants: to grow or defend. Q Rev Biol 67:283–335. doi: 10.1086/417659 CrossRefGoogle Scholar
  21. Horigome T, Kumar R, Okamoto K (1988) Effects of condensed tannins prepared from fodder plant on digestive enzymes in vitro and in the intestine of rats. Br J Nutr 60:275–285CrossRefPubMedGoogle Scholar
  22. Hothorn T, Bretz F, Westfall P (2008) Simultaneous inference in general parametric models. Biom J 50:346–363CrossRefPubMedGoogle Scholar
  23. Janzen DH (1974) Tropical black water rivers, animals and mast fruiting by the Dipterocarpaceae. Biotropica 6:69–103. doi: 10.2307/2989823 CrossRefGoogle Scholar
  24. Knowles BH (1994) Mechanism of action of Bacillus thuringiensis insecticidal delta-endotoxins. Adv Ins Phys 24(24):275–308. doi: 10.1016/s0065-2806(08)60085-5 Google Scholar
  25. Koptur S (1985) Alternative defenses against herbivores in Inga (Fabaceae: Mimosoideae) over an elevational gradient. Ecology 66:1639–1650CrossRefGoogle Scholar
  26. Kumar R, Horigome T (1986) Fractionation, characterization and protein-precipitating capacity of the condensed tannins from Robinia pseudoacacia L. leaves. J Agric Food Chem 34:487–489CrossRefGoogle Scholar
  27. Lokvam J, Kursar TA (2005) Divergence in structure and activity of phenolic defenses in two co-occurring Inga species. J Chem Ecol 31:2563–2580CrossRefPubMedGoogle Scholar
  28. Martin MM, Martin JS (1984) Surfactants—their role in preventing the precipitation of proteins by tannins in insect guts. Oecologia 61:342–345. doi: 10.1007/bf00379632 CrossRefGoogle Scholar
  29. Misiewicz TM, Fine PVA (2014) Evidence for ecological divergence across a mosaic of soil types in an Amazonian tropical tree: Protium subserratum (Burseraceae). Mol Ecol 23:2543–2558. doi: 10.1111/mec.12746 CrossRefPubMedGoogle Scholar
  30. R_Development_Core_Team (2013) R: A language and environment for statistical computing. Computing RFfS, Vienna, AustriaGoogle Scholar
  31. Ruokolainen K, Tuomisto H (1998) Tropical forests are not flat: how mountains affect herbivore diversity. Ecol Let 13:1348–1357Google Scholar
  32. Salminen JP, Karonen M (2011) Chemical ecology of tannins and other phenolics: we need a change in approach. Funct Ecol 25:325–338. doi: 10.1111/j.1365-2435.2010.01826.x CrossRefGoogle Scholar
  33. Strumeyer DH, Malin MJ (1975) Condensed tannins in grain-sorghum—isolation, fractionation, and characterization. J Agric Food Chem 23:909–914. doi: 10.1021/jf60201a019 CrossRefPubMedGoogle Scholar
  34. Sugiyama H, Akazome Y, Shoji T, Yamaguchi A, Yasue M, Kanda T, Ohtake Y (2007) Oligomeric procyanidins in apple polyphenol are main active components for inhibition of pancreatic lipase and triglyceride absorption. J Agric Food Chem 55:4604–4609. doi: 10.1021/jf070569k CrossRefPubMedGoogle Scholar
  35. Swain T (1979) Tannins and lignins. In: Rosenthal GA, Janzen DH (eds) Herbivores: their interactions with secondary plant metabolites. Academic Press, New York, pp 657–682Google Scholar
  36. Thompson RS, Jacques D, Haslam E, Tanner RJN (1972) Plant proanthocyanidins. Part 1. Introduction; the isolation, structure, and distribution in nature of plant procyanidins. J Chem Soc Perk Trans 1:1387–1399CrossRefGoogle Scholar

Copyright information

© Springer Basel 2015

Authors and Affiliations

  1. 1.Department of Integrative BiologyUniversity of CaliforniaBerkeleyUSA
  2. 2.Department of BiologyLewis & Clark CollegePortlandUSA
  3. 3.Agricultural Research Service, United States Department of AgricultureAlbanyUSA

Personalised recommendations