Abstract
Bark stripping by mammals is a major problem in managed conifer forests worldwide. In Australia, bark stripping in the exotic plantations of Pinus radiata is mainly caused by native marsupials and results in reduced survival, growth, and in extreme cases death of trees. Herbivory is influenced by a balance between primary metabolites that are sources of nutrition and secondary metabolites that act as defences. Identifying the compounds that influence herbivory may be a useful tool in the management of forest systems. This study aimed to detect and identify both constitutive and induced compounds that are associated with genetic differences in susceptibility of two-year-old P. radiata trees to bark stripping by marsupials. An untargeted profiling of 83 primary and secondary compounds of the needles and bark samples from 21 susceptible and 21 resistant families was undertaken. These were among the most and least damaged families, respectively, screened in a trial of 74 families that were exposed to natural field bark stripping by marsupials. Experimental plants were in the same field trial but protected from bark stripping and a subset were subjected to artificial bark stripping to examine induced and constitutive chemistry differences between resistant and susceptible families. Machine learning (random forest), partial least squares plus discriminant analysis (PLS-DA), and principal components analysis with discriminant analysis (PCA-DA), as well as univariate methods were used to identify the most important totals by compound group and individual compounds differentiating the resistant and susceptible families. In the bark, the constitutive amount of two sesquiterpenoids — bicyclogermacrene and an unknown sesquiterpenoid alcohol —were shown to be of higher levels in the resistant families, whereas the constitutive sugars, fructose, and glucose, as well individual phenolics, were higher in the more susceptible families. The chemistry of the needles was not useful in differentiating the resistant and susceptible families to marsupial bark stripping. After artificial bark stripping, the terpenes, sugars, and phenolics responded in both the resistant and susceptible families by increasing or reducing amounts, which leveled the differences in the amounts of the compounds between the different resistant and susceptible classes observed at the constitutive level. Overall, based on the families with extreme values for less and more susceptibility, differences in the amounts of secondary compounds were subtle and susceptibility due to sugars may outweigh defence as the cause of the genetic variation in bark stripping observed in this non-native tree herbivory system.
Similar content being viewed by others
Availability of data and material
Not applicable.
Code availability
Not applicable.
References
Achotegui-Castells A, Llusià J, Hódar JA, Peñuelas J (2013) Needle terpene concentrations and emissions of two coexisting subspecies of Scots pine attacked by the pine processionary moth (Thaumetopoea pityocampa). Acta Physiol Plant 35:3047–3058
Agrawal AA, Weber MG (2015) On the study of plant defence and herbivory using comparative approaches: how important are secondary plant compounds. Ecol Lett 18:985–991
Amri I, Hanana M, Gargouri S, Jamoussi B, Hamrouni L (2013) Comparative study of two coniferous species (Pinus pinaster Aiton and Cupressus sempervirens L. var. dupreziana [A. Camus] Silba) essential oils: chemical composition and biological activity. Chilean Journal of Agricultural Research 73:259–266
Boyle RR (1999) The metabolic fate of dietary terpenes in folivorous marsupials. University of Tasmania, Hobart
Bredsdorff L, Wedebye EB, Nikolov NG, Hallas-Møller T, Pilegaard K (2015) Raspberry ketone in food supplements – High intake, few toxicity data – A cause for safety concern? Regul Toxicol Pharmacol 73:196–200
Breiman L (2001) Random forests. Mach Learn 45:5–32
Bucyanayandi JD, Bergeron J-M, Menard H (1990) Preference of meadow voles (Microtus pennsylvanicus) for conifer seedlings: chemical components and nutritional quality of bark of damaged and undamaged trees. J Chem Ecol 16:2569–2579
Butler DG, Cullis BR, Gilmour AR, Gogel BJ (2009) ASReml-R reference manual. In. ' Version 3 edn. pp. 149. (Queensland Government Department of Primary Industries and Fisheries: Brisbane, Qld)
Carmona D, Lajeunesse MJ, Johnson MTJ (2011) Plant traits that predict resistance to herbivores. Funct Ecol 25:358–367
Celedon JM, Bohlmann J (2019) Oleoresin defenses in conifers: chemical diversity, terpene synthases and limitations of oleoresin defense under climate change. New Phytol 224:1444–1463
Chen T, Cao Y, Zhang Y, Liu J, Bao Y, Wang C, Jia W, Zhao A (2013) Random forest in clinical metabolomics for phenotypic discrimination and biomarker selection. Evidence-Based Complementary and Alternative Medicine 2013:11
Clancy KM (1992) The role of sugars in western spruce budworm nutritional ecology. Ecological Entomology 17:189–197
Danielsson M, Lundén K, et al. (2011) Chemical and transcriptional responses of Norway spruce genotypes with different susceptibility to Heterobasidion spp. infection. BMC Plant Biology 11, 154.
DPIPWE (2019) Annual state-wide spotlight surveys, Tasmania 2018/19. Regional summary: priority harvested species. In. ' Ed. DoPIPWa Environment). (Tasmanian Government: Tasmania)
El-Merhibi A, Ngo SNT, Jones BR, Milic NL, Stupans I, McKinnon RA (2007) Molecular insights into xenobiotic disposition in Australian marsupials. Australasian Journal of Ecotoxicology 13:7–18
Farentinos RC, Capretta PJ, Kepner RE, Littlefield VM (1981) Selective herbivory in tassel-eared squirrels: role of monoterpenes in ponderosa pines chosen as feeding trees. Science 213:1273–1275
Felicijan M, Novak M, Kraševec N, Urbanek Krajnc A (2015) Antioxidant defences of Norway spruce bark against bark beetles and its associated blue-stain fungus. Agricultura 12:9–18
Franceschi VR, Krokene P, Christiansen E, Krekling T (2005) Anatomical and chemical defenses of conifer bark against bark beetles and other pests. New Phytol 167:353–376
Ganthaler A, Stöggl W, Kranner I, Mayr S (2017) Foliar phenolic compounds in Norway spruce with varying susceptibility to Chrysomyxa rhododendri: analyses of seasonal and infection-induced accumulation patterns. Frontiers in Plant Science 8.
Gershenzon J (1994) Metabolic costs of terpenoid accumulation in higher plants. J Chem Ecol 20:1281–1328
Gill RMA (1992) A review of damage by mammals in north temperate forests: 1 Deer. Forestry 65:145–169
Goodsman DW, Lusebrink I, Landhäusser SM, Erbilgin N, Lieffers VJ (2013) Variation in carbon availability, defense chemistry and susceptibility to fungal invasion along the stems of mature trees. New Phytol 197:586–594
Gromski PS, Muhamadali H, Ellis DI, Xu Y, Correa E, Turner ML, Goodacre R (2015) A tutorial review: Metabolomics and partial least squares-discriminant analysis–a marriage of convenience or a shotgun wedding. Anal Chim Acta 879:10–23
Hammerbacher A, Ralph SG, Bohlmann J, Fenning TM, Gershenzon J, Schmidt A (2011) Biosynthesis of the Major Tetrahydroxystilbenes in Spruce, Astringin and Isorhapontin, Proceeds via Resveratrol and Is Enhanced by Fungal Infection. Plant Physiol 157:876
Hansson L, Gref R, Lundren L, Theander O (1986) Susceptibility to vole attacks due to bark phenols and terpenes in Pinus contorta provenances introduced into Sweden. J Chem Ecol 12:1569–1578
Hummelbrunner LA, Isman MB (2001) Acute, sublethal, antifeedant, and synergistic effects of monoterpenoid essential oil compounds on the tobacco cutworm, Spodoptera litura (Lep., Noctuidae). J Agric Food Chem 49:715–720
Iason G, Palo R (1991) Effects of birch phenolics on a grazing and a browsing mammal: a comparison of hares. J Chem Ecol 17:1733–1743
Iason GR, O’Reilly-Wapstra JM, Brewer MJ, Summers RW, Moore BD (2011) Do multiple herbivores maintain chemical diversity of Scots pine monoterpenes? Philosophical Transactions of the Royal Society b: Biological Sciences 366:1337–1345
Johnson KS, Felton GW (2001) Plant phenolics as dietary antioxidants for herbivorous insects: A test with genetically modified tobacco. J Chem Ecol 27:2579–2597
Jombart T, Collins C (2015) A tutorial for discriminant analysis of principal components (DAPC) using adegenet 2.0.0. In. ’. London, Imperial College London, MRC Centre for Outbreak Analysis and Modelling. http://adegenet.r-forge.r-project.org/files/tutorial-dapc.pdf. Accessed 20 Nov 2020
Keeling CI, Bohlmann J (2006) Genes, enzymes and chemicals of terpenoid diversity in the constitutive and induced defence of conifers against insects and pathogens. New Phytol 170:657–675
Kimball BA, Nolte DL, Engeman RM, Johnston JJ, Stermitz FR (1998) Chemically mediated foraging preference of black bears (Ursus americanus). J Mammal 79:448–456
Kurek T, Todys J, Pazdrowski W, Szymański M, Łukowski A (2019) Intensity of stripping and sugar content in the bark and the bast of European beech (Fagus sylvatica). Open Life Sciences 14:19–28
Lebouvier N, Hue T, Hnawia E, Lesaffre L, Menut C, Nour M (2013) Acaricidal activity of essential oils from five endemic conifers of New Caledonia on the cattle tick Rhipicephalus (Boophilus) microplus. Parasitol Res 112:1379–1384
Lee SY, Mediani A, Maulidiani M, Khatib A, Ismail IS, Zawawi N, Abas F (2018) Comparison of partial least squares and random forests for evaluating relationship between phenolics and bioactivities of Neptunia oleracea. J Sci Food Agric 98:240–252
Li C-Y, Weiss D, Goldschmidt EE (2003) Girdling affects carbohydrate-related gene expression in leaves, bark and roots of alternate-bearing citrus trees. Ann Bot 92:137–143
Lindroth RL, Batzli GO (1984) Plant phenolics as chemical defenses: effects of natural phenolics on survival and growth of prairie voles (Microtus ochrogaster). J Chem Ecol 10:229–244
López-Goldar X, Villari C, Bonello P, Borg-Karlson A-K, Grivet D, Zas R, Sampedro L (2018) Inducibility of plant secondary metabolites in the stem predicts genetic variation in resistance against a key insect herbivore in maritime pine. Frontiers in Plant Science 9.
Lundborg L (2016) Effects of methyl jasmonate on chemical defenses of conifer seedlings in relation to feeding by Hylobius abietis. Doctoral thesis Thesis, KTH Royal Institute of Technology, Stockholm
Madan SS, Wasewar KL, Pandharipande SL (2016) Modeling the adsorption of benzeneacetic acid on CaO2 nanoparticles using artificial neural network. Resource-Efficient Technologies 2:S53–S62
Mead DJ (2013) Sustainable management of Pinus radiata plantations. Rome, Food and Agriculture Organization of the United Nations (FAO)
Mendez KM, Reinke SN, Broadhurst DI (2019) A comparative evaluation of the generalised predictive ability of eight machine learning algorithms across ten clinical metabolomics data sets for binary classification. Metabolomics 15:150
Miller A, O'Reilly-Wapstra J, Potts B (2014) Genetic variation in bark stripping among Pinus radiata. National Centre for Future Forest Industries, Hobart.
Moore BD, DeGabriel JL (2012) Integrating the effects of PSMs on vertebrate herbivores across spatial and temporal scales. In 'The ecology of plant secondary metabolites: from genes to global processes. Vol. 226. (Eds GR Iason, M Dicke and SE Hartley). (Cambridge University Press: Cambridge).
Moreira X, Lundborg L, Zas R, Carrillo-Gavilán A, Borg-Karlson A-K, Sampedro L (2013a) Inducibility of chemical defences by two chewing insect herbivores in pine trees is specific to targeted plant tissue, particular herbivore and defensive trait. Phytochemistry 94:113–122
Moreira X, Zas R, Sampedro L (2013b) Additive genetic variation in resistance traits of an exotic pine species: little evidence for constraints on evolution of resistance against native herbivores. Heredity 110:449–456
Moreira X, Zas R, Sampedro L (2012) Differential allocation of constitutive and induced chemical defenses in pine tree juveniles: a test of the optimal defense theory. PLoS One 7, e34006.
Morkunas I, Ratajczak L (2014) The role of sugar signaling in plant defense responses against fungal pathogens. Acta Physiol Plant 36:1607–1619
Mumm R, Hilker M (2006) Direct and indirect chemical defence of pine against folivorous insects. Trends Plant Sci 11:351–358
Nantongo JS, Potts BM, Fitzgerald H, Newman J, Elms S, Aurik D, Dungey H, O’Reilly-Wapstra JM (2020) Quantitative genetic variation in bark stripping of Pinus radiata. Forests 11:1356
Nantongo JS, Potts B, Rodemann T, Fitzgerald H, Davies N, O’Reilly-Wapstra J (2021a) Developing near infrared spectroscopy models for predicting chemistry and responses to stress in Pinus radiata (D. Don). Journal of Near Infrared Spectroscopy, 09670335211006526.
Nantongo JS, Potts BM, Rodemann T, O'Reilly-Wapstra JM (2021b) Linking genetics and chemistry to minimise bark stripping in Pinus radiata. PhD Thesis Thesis, University of Tasmania,
O’Reilly-Wapstra JM, Iason GR, Thoss V (2007) The role of genetic and chemical variation of Pinus sylvestris seedlings in influencing slug herbivory. Oecologia 152:82–91
Page DE, Close D, Beadle CL, Wardlaw TJ, Mohammed CL (2013) Seasonal dynamics in understorey abundance and carbohydrate concentration in relation to browsing and bark stripping of Tasmanian Pinus radiata plantations. For Ecol Manage 296:98–107
Pederson JC, Welch BL (1985) Comparison of ponderosa pines as feed and nonfeed trees for Abert squirrels. J Chem Ecol 11:149–157
Phillips MA, Croteau RB (1999) Resin-based defenses in conifers. Trends Plant Sci 4:184–190
Preda C, Saporta G, Lévéder C (2007) PLS classification of functional data. Comput Statistics 22:223–235
R Core Team (2018) R: A language and environment for statistical computing. In. ’. Vienna, Austria, R Foundation for Statistical Computing
Radwan MA (1972) Differences between Douglas-fir genotypes in relation to browsing preference by black-tailed deer. Can J for Res 2:250–255
Radwan MA, Crouch GL (1978) Selected chemical constituents and deer browsing preference of Douglas fir. J Chem Ecol 4:675–683
Raffa K (2014) Terpenes tell different tales at different scales: glimpses into the chemical ecology of conifer - bark beetle - microbial interactions. J Chem Ecol 40:1–20
Raffa KF, Smalley EB (1995) Interaction of pre-attack and induced monoterpene concentrations in host conifer defense against bark beetle-fungal complexes. Oecologia 102:285–295
Raffa KF, Mason CJ, Bonello P, Cook S, Erbilgin N, Keefover-Ring K, Klutsch JG, Villari C, Townsend PA (2017) Defense syndromes in lodgepole – whitebark pine ecosystems relate to degree of historical exposure to mountain pine beetles. Plant, Cell Environ 40:1791–1806
Rea RV, Hjeljord O, Härkönen S (2014) Differential selection of North American and Scandinavian conifer browse by northwestern moose (Alces alces andersoni) in winter. Acta Theriol 59:353–360
Reglinski T, Taylor JT, Northcott GL, Ah Chee A, Spiers M, Wohlers M, Hill RA (2017) Biochemical responses associated with induced resistance to Colletotrichum acutatum in Pinus radiata seedlings treated with methyl jasmonate and Trichoderma spp. Forest Pathology 47, e12350.
Roitto M, Rautio P, Markkola A, Julkunen-tiitto R, Varama M, Saravesi K, Tuomi J (2009) Induced accumulation of phenolics and sawfly performance in Scots pine in response to previous defoliation. Tree Physiol 29:207–216
Roth M, Hussain A, Cale JA, Erbilgin N (2018) Successful colonization of lodgepole pine trees by mountain pine beetle increased monoterpene production and exhausted carbohydrate reserves. J Chem Ecol 44:209–214
Saccenti E, Hoefsloot HCJ, Smilde AK, Westerhuis JA, Hendriks MMWB (2014) Reflections on univariate and multivariate analysis of metabolomics data. Metabolomics 10:361–374
Saint-Andrieux C, Bonenfant C, Toïgo C, Basille M, Klein F (2009) Factors affecting beech Fagus sylvatica bark stripping by red deer Cervus elaphus in a mixed forest. Wildl Biol 15:187–197
Sauve DG, Cote SD (2007) Winter forage selection in white-tailed deer at high density: balsam fir is the best of a bad choice. J Wildl Manag 71:911–914
Scalerandi E, Flores GA, Palacio M, Defagó MT, Carpinella MC, Valladares G, Bertoni A, Palacios SM (2018) Understanding synergistic toxicity of terpenes as insecticides: contribution of metabolic detoxification in Musca domestica. Front Plant Sci 9:1579
Schowalter TD (2012) Ecology and management of bark beetles (Coleoptera: Curculionidae: Scolytinae) in southern pine forests. Journal of Integrated Pest Management 3:A1–A7
Schwachtje J, Baldwin IT (2008) Why does herbivore attack reconfigure primary metabolism? Plant Physiol 146:845–851
Seybold SJ, Huber DPW, Lee JC, Graves AD, Bohlmann J (2006) Pine monoterpenes and pine bark beetles: a marriage of convenience for defense and chemical communication. Phytochem Rev 5:143–178
Smith AH, Ratkowsky DA, Wardlaw TJ, Mohammed CL (2020) Ease of access to an alternative food source enables wallabies to strip bark in Tasmanian Pinus radiata plantations. Forests 11:387
Snyder MA (1992) Selective herbivory by Abert’s squirrel mediated by chemical variability in ponderosa pine. Ecology 73:1730–1741
Stutz RS, Croak BM, Proschogo N, Banks PB, McArthur C (2017) Olfactory and visual plant cues as drivers of selective herbivory. Oikos 126:259–268
Sunnerheim-Sjöberg K, Hämäläinen M (1992) Multivariate study of moose browsing in relation to phenol pattern in pine needles. J Chem Ecol 18:659–672
Tamura N, Ohara S (2005) Chemical components of hardwood barks stripped by the alien squirrel Callosciurus erythraeus in Japan. J for Res 10:429–433
Vinaixa M, Samino S, Saez I, Duran J, Guinovart JJ, Yanes O (2012) A guideline to univariate statistical analysis for LC/MS-based untargeted metabolomics-derived data. Metabolites 2:775–795
Vourc’h G, De Garine-Wichatitsky M, Labbé A, Rosolowski D, Martin J-L, Fritz H (2002a) Monoterpene effect on feeding choice by deer. J Chem Ecol 28:2411–2427
Vourc’h G, Vila B, Gillon D, Escarré J, Guibal F, Fritz H, Clausen TP, Martin JL (2002b) Disentangling the causes of damage variation by deer browsing on young Thuja plicata. Oikos 98:271–283
Westoby M (1978) What are the biological bases of varied diets? Am Nat 112:627–631
Whitehill JGA, Yuen MMS, Henderson H, Madilao L, Kshatriya K, Bryan J, Jaquish B, Bohlmann J (2019) Functions of stone cells and oleoresin terpenes in the conifer defense syndrome. New Phytol 221:1503–1517
Wiley E, Helliker B (2012) A re-evaluation of carbon storage in trees lends greater support for carbon limitation to growth. New Phytol 195:285–289
Wright MN, Ziegler A (2015) ranger: A fast implementation of random forests for high dimensional data in C++ and R. J Stat Softw 77(1508):04409
Zhang X, States JS (1991) Selective herbivory of ponderosa pine by Abert squirrels: a re-examination of the role of terpenes. Biochem Syst Ecol 19:111–115
Zou J, Cates RG (1994) Role of Douglas fir (Pseudotsuga menziesii) carbohydrates in resistance to budworm (Choristoneura occidentalis). J Chem Ecol 20:395–405
Acknowledgements
We thank industrial partners, Timberlands Pacific Pty Ltd and the Radiata Pine Breeding Company for the provision of genetic material. We also that Hancock Victorian Plantations and Scion for their support of the project. Judith Ssali Nantongo also acknowledges receipt of a Tasmania Graduate Research Scholarship.
Funding
Funding for this project was under Australian Research Council (ARC) Linkage Grant LP140100602.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Ethics approval
The experimental work for the field trials in this thesis was performed with the approval of the University of Tasmania Animal Ethics Committee (Permit No. A0015577).
Consent for publication
All the Authors approved the publication of the manuscript.
Conflicts of interest/Competing interests
The authors declare no conflict of interest.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Nantongo, J.S., Potts, B.M., Davies, N.W. et al. Chemical Traits that Predict Susceptibility of Pinus radiata to Marsupial Bark Stripping. J Chem Ecol 48, 51–70 (2022). https://doi.org/10.1007/s10886-021-01307-5
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10886-021-01307-5