Profiling Secondary Metabolites of Needles of Ozone-Fumigated White Pine (Pinus strobus) Clones by Thermally Assisted Hydrolysis/Methylation GC/MS
- 181 Downloads
Plant secondary metabolites have an important role in defense responses against herbivores and pathogens, and as a chemical barrier to elevated levels of harmful air pollutants. This study involves the rapid chemical profiling of phenolic and diterpene resin acids in needles of two (ozone-tolerant and ozone-sensitive) white pine (Pinus strobus) clones, fumigated with different ozone levels (control, and daily events peaking at 80 and 200 ppb) for 40 days. The phenolic and resin acids were measured using thermally assisted hydrolysis and methylation (THM) gas chromatography/mass spectrometry. Short-term fumigation affected the levels of two phenolic acids, i.e., 3-hydroxybenzoic and 3,4-dihydroxybenzoic acids, in that both showed a substantial decrease in concentration with increased ozone dose. The decrease in concentration of these THM products may be caused by inhibition of the plant’s shikimate biochemical pathway caused by ozone exposure. The combined occurrence of these two ozone-sensitive indicators has a role in biomonitoring of ozone levels and its impact on forest productivity. In addition, chromatographic profile differences in the major diterpene resin acid components were observed between ozone-tolerant and ozone-sensitive clones. The resin acids anticopalic, 3-oxoanticopalic, 3β-hydroxyanticopalic, and 3,4-cycloanticopalic acids were present in the ozone-sensitive pine; however, only anticopalic acid was present in the ozone-tolerant clone. This phenotypic variation in resin acid composition may be useful in distinguishing populations that are differentially adapted to air pollutants.
KeywordsOzone White pine (Pinus strobus) Needles Secondary metabolites Thermally assisted hydrolysis and methylation GC/MS Phenolic acids Diterpene resin acids
The authors thank Frontier Laboratories (Japan) for allowing the use of the vertical microfurnace pyrolyzer. Funding from NSERC and Memorial University is acknowledged.
- Charland, M., Malcolm, J. W., and Cox, R. M. 1994. Effects of Ozone and SO2 on Eastern White Pine Genotypes. In: Air Pollution and Multiple Stresses. Proc. 16th International Meeting for Specialists in Air Pollution Effects on Forest Ecosystems. (R. Cox, K. Percy, K. Jensen, and C. Simpson, Compilers) Canadian Forest Service-Atlantic Forestry Centre, Natural Resources Canada, Fredericton, N. B. p. 402.Google Scholar
- Charland M., Malcolm, J. W., and Cox, R. M. 1995. In: Photosynthesis: from light to biosphere, Proc. of the 10th International Photosynthesis Congress, Montpellier, Fr., Aug. 20–25, 1995, Vol. 5, Mathis, P., Kluwer (Eds). Dordrecht, Netherlands, pp. 921–924.Google Scholar
- Eckey-Kalthenback, H., Ernest, D., Heller, W., and Sandermann, H. Jr. 1994. Biochemical plant responses to ozone. IV. Cross-induction of defensive pathways in parsley (Petroselinum crispum) plants. Plant Physiol. 104:67–74.Google Scholar
- Kicinski, H. S., Kettrup, A., Boos, K. S., and Masuch, G. 1988. Single and combined effects of continuous and discontinuous O3 and SO2 immersion of Norway spruce needles. Int. J. Environ. Anal. Chem. 32:213–241.Google Scholar
- Langebartels, C., Heller, W., Kerner, K., Leonard, S., Rosemann, D., Schraudner, M., Trost, M., and Sandermann, H. Jr. 1990. Ozone-induced defense reactions in plants, pp. 358-368, in Pfirrmann, T. and Payer H. (Eds.) Environmental Research with Plants in Closed Chambers. Neuherberg: Commission of European Communities Air Pollution Report.Google Scholar
- Madonna, A. J., and Voorhees, K. J., Hadfield, T. L. 2001. Rapid detection of taxonomically important fatty acid methyl ester and steroid biomarkers using in-situ thermal hydrolysis/methylation mass spectrometry (THM-MS): implications for bioaerosol detection. J. Anal. Appl. Pyrolysis 61:65–89.CrossRefGoogle Scholar
- Manninen, A.-M. 1999. Susceptibility of Scots pine seedlings to specialist and generalist insect herbivores: Importance of plant defence and mycorrhizal status. Kuopio Univ. Publications C. Nat. Environ Sci. 100.Google Scholar
- Skelly, J. M., Chappelka, A. H., Laurence, J. A., and Fredericksen, T. S. 1997. in Sandermann, H., Welburn, A. and health R. (Eds.). Forest decline and Ozone: a Comparison of Controlled Chamber and Field Experiments (Ecological Studies Vol. 127) Springer, New York.Google Scholar
- Watts, S., and De Al Rie, E. R. 2002. GCMS analysis of triterpenoid resins: in situ derivatization procedures using quaternary ammonium hydroxides. Stud. Conserv. 47:257–272.Google Scholar
- Zhang, H., 1993. The Analysis of Organic Constituents in Leaves by Pyrolytic-Gas Chromatography and its Application to Selected Environmental Effects on Plants. M.Sc. Dissertation, Memorial University of Newfoundland, St. John’s, Canada, p. 136.Google Scholar