Skip to main content
Log in

Application of metabolomics to genotype and phenotype discrimination of birch trees grown in a long-term open-field experiment

  • Original Article
  • Published:
Metabolomics Aims and scope Submit manuscript

Abstract

The principal objectives of the study were to develop metabolomics tool and to test its efficiency for discrimination and biochemical pattern recognition of genotypes and phenotypes of silver birch trees (Betula pendula Roth). In the experiment were used two birch genotypes (GT 2 and GT 5) that have been grown over seven years on the two open fields A and B. The metabolomics tool was applied also to study biochemical responses of the GT 2 to elevated (1.5 × ambient) concentration of ozone as an environmental stress factor. These trees were treated with ozone over seven years using an open-air exposure system. The developed metabolomics tool was based on the analyses of lipophilic and polar compounds of birch leaves with GC-MS and HPLC-DAD (polar phenolics only). The metabolome database included 331 chemical traits and was analyzed with descriptive and multivariate statistics. Application of cluster and principle component analyses clearly discriminated genetically different birch trees. In addition, the genotype clusters were further divided into two subclusters corresponding to trees from field A and field B. Formation of these phenotypes was due to the differences in some environmental conditions between the field A and field B. Biochemical discrimination between phenotypes of control and ozone-treated birch trees of GT 2 was found also. However, distances between these phenotypes, as well as between phenotypes of control trees from the field A and field B were found to be considerably smaller than between birch genotypes. Metabolites with the largest contribution to birch genotype/phenotype discrimination were determined and some were identified.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Choi, H.-K., Choi, Y., Verberne, M., Lefeber, A. W. M., Erkelens, C., & Verpoorte, R. (2004). Metabolic fingerprinting of wild type and transgenic tobacco plants by 1H NMR and multivariate analysis technique. Phytochemistry, 65, 857–864.

    Article  PubMed  CAS  Google Scholar 

  • Daub, C. O., Kloska, S., & Selbig, J. (2003). MetaGeneAlyse: Analysis of integrated transcriptional and metabolite data. Bioinformatics, 19, 2332–2333.

    Article  PubMed  CAS  Google Scholar 

  • Fiehn, O. (2002). Metabolomics––the link between genotypes and phenotypes. Plant Molecular Biology, 48, 155–171.

    Article  PubMed  CAS  Google Scholar 

  • Fiehn, O., Kopka, J., Dörmann, P., Altman, T., Trethewey, R., & Willmitzer, L. (2000). Metabolite profiling for functional genomics. Nature Biotechnology, 18, 1157–1161.

    Article  PubMed  CAS  Google Scholar 

  • Halket, J., Przyborowska, A., Stein, S., Down, S., & Chalmers, R. (1999). Deconvolution gas chromatography/mass spectrometry of urinary organic acid––potential for pattern recognition and automated identification of metabolite disorders. Rapid Communications in Mass Spectrometry, 13, 279–284.

    Article  PubMed  CAS  Google Scholar 

  • Haukioja, E., Ossipov, V., Koricheva, J., Honkanen, T., Larsson, S., & Lempa, K. (1998). Biosynthetic origin of carbon-based secondary compounds: Cause of variable responses of woody plants to fertilization? Chemoecology, 8, 133–139.

    Article  CAS  Google Scholar 

  • Ingestad, T. (1962). Macro element nutrition of pine, spruce and birch seedling in nutrient solutions. Meddelande Statens Skogsforskningsinstitut, 51, 1–133.

    Google Scholar 

  • Jolliffe, I. T. (1986). Principal component analysis. New York: Springer-Verlag.

    Google Scholar 

  • Karnosky, D. F., Werner, H., Holopainen, T., Percy, K., Oksanen, T., Oksanen, E., Heerdt, C., Fabian, P., Nagy, J., Heilman, W., Cox, R., Nelson, N., & Matyssek, R. (2007). Free-air exposure systems to scale up ozone research to mature trees. Plant Biology, 9, 181–190.

    Article  PubMed  CAS  Google Scholar 

  • de Kroone, H., Huber, H., Stuefer, J., & van Groenendael, J. (2005). A modular concept of phenotypic plasticity in plants. The New Phytologist, 166, 73–82.

    Article  Google Scholar 

  • Kontunen-Soppela, S., Ossipov, V., Ossipova, S., & Oksanen, E. (2007). Shift in birch leaf metabolome and carbon allocation during long-term open-field ozone exposure. Global Change Biology, 13, 1053–1067.

    Article  Google Scholar 

  • Laurila, T., Tuovinen, J.-P., Tarvainen, V., & Simpson, D. (2004). Trends and scenarios of ground-level ozone concentrations in Finland. Boreal Environment Research, 9, 167–184.

    Google Scholar 

  • Lavola, A., Julkunen-Tiitto, R., & Pääkkönen, E. (1994). Does ozone stress change the primary or secondary metabolites of birch (Betula pendula Roth)? The New Phytologist, 126, 637–642.

    Article  CAS  Google Scholar 

  • Loponen, J., Ossipov, V., Lempa, K., Haukioja, E., & Pihlaja, K. (1998). Concentrations and among-compound correlations of individual phenolics in white birch leaves under air pollution stress. Chemosphere, 36, 1445–1456.

    Article  Google Scholar 

  • Oksanen, E. (2003). Responses of selected birch (Betula pendula). clones to ozone change over time. Plant, Cell & Environment, 26, 875–886.

    Article  Google Scholar 

  • Ossipov, V., Haukioja, E., Ossipova, S., Hanhimäki, S., & Pihlaja, K. (2001). Phenolic and phenolic-related factors as determinants of suitability of mountain birch leaves to an herbivorous insect. Biochemical Systematics and Ecology, 29, 223–240.

    Article  PubMed  CAS  Google Scholar 

  • Ossipov, V., Nurmi, K., Loponen, J., Haukioja, E., & Pihlaja, K. (1996). HPLC separation and identification of phenolic compounds from leaves of Betula pubescens and Betula pendula. Journal of Chromatography. A, 721, 59–68.

    Article  CAS  Google Scholar 

  • Ossipov, V., Nurmi, K., Loponen, J., Prokopiev, N., Haukioja, E., & Pihlaja, K. (1995). HPLC isolation and identification of flavonoids from white birch Betula pubescens leaves. Biochemical Systematics and Ecology, 23, 213–222.

    Article  CAS  Google Scholar 

  • Ossipova S., Ossipov, V., Haukioja, E., Loponen, J., & Pihlaja, K. (2001). Proanthocyanidins from mountain birch leaves: Quantitation and properties. Phytochemical Analysis, 12, 128–133.

    Article  PubMed  CAS  Google Scholar 

  • Pell, E. J., Schlagnhaufer, C. D., & Arteca, R. N. (1997). Ozone-induced oxidative stress: mechanisms of action and reaction. Physiologia Plantarum, 100, 264–273.

    Article  CAS  Google Scholar 

  • Peltonen, P. A., Vapaavuori, E., & Julkunen-Tiitto, R. (2005). Accumulation of phenolic compounds in birch leaves is changed by elevated carbon dioxide and ozone. Global Change Biology, 11, 1305–1324.

    Article  Google Scholar 

  • Riikonen, J., Lindsberg, M.-M., Holopainen, T., Oksanen, E., Lappi, J., Peltonen, P., & Vapaavuori, E. (2004). Silver birch and climate change: variable growth and carbon allocation responses to elevated concentrations of carbon dioxide and ozone. Tree Physiology, 24, 1227–1237.

    PubMed  CAS  Google Scholar 

  • Roessner, U., Luedemann, A., Brust, D., Fiehn, O., Linke, T., Willmitzer, L., & Fernie, A. (2001). Metabolic profiling allows comprehensive phenotyping of genetically or environmentally modified plant system. Plant Cell, 13, 11–29.

    Article  PubMed  CAS  Google Scholar 

  • Saleem, A., Loponen, J., Pihlaja, K., & Oksanen, E. (2001). Effects of long-term open-field ozone exposure on leaf phenolics of European silver birch (Betula pendula Roth). Journal of Chemical Ecology, 27, 1049–1062.

    Article  PubMed  CAS  Google Scholar 

  • Salminen, J.-P., Ossipov, V., Loponen, J., Haukioja, E., & Pihlaja, K. (1999). Characterisation of hydrolysable tannins from leaves of Betula pubescens by high-performance liquid chromatography––mass spectrometry. Journal of Chromatography. A, 864, 283–291.

    Article  CAS  Google Scholar 

  • Sumner, L., Mendes, P., & Dixon, R. (2003). Plant metabolomics: Large-scale phytochemistry in the functional genomics era. Phytochemistry, 62, 817–836.

    Article  PubMed  CAS  Google Scholar 

  • Taylor, J., King, R. D., Altmann, T., & Fiehn, O. (2002). Application of metabolomics to plant genotype discrimination using statistics and machine learning. Bioinformatics, 18, 241–248.

    Google Scholar 

  • Trethewey, R. (2001). Gene discovery via metabolic profiling. Current Opinion in Biotechnology, 12, 135–138.

    Article  PubMed  CAS  Google Scholar 

  • Vinocur, B., & Altman, A. (2005). Recent advances in engineering plant tolerance to abiotic stress: Achievements and limitations. Current Opinion in Biotechnology, 16, 123–132.

    Article  PubMed  CAS  Google Scholar 

  • Valkama, E., Koricheva, J., Ossipov, V., Ossipova, S., Haukioja, E., & Pihlaja, K. (2005). Delayed induced responses of birch glandular trichomes and leaf surface lipophilic compounds to mechanical defoliation and simulated browsing. Oecologia, 146, 385–393.

    Article  PubMed  Google Scholar 

  • Valkama, E., Koricheva, J., & Oksanen, E. (2007). Effects of elevated O3, alone and in combination with elevated CO2, on tree leaf chemistry and insect herbivore performance: A meta-analysis. Global Change Biology, 13, 184–201.

    Article  Google Scholar 

  • Wang, W., Vinocur, B., Shoseyov, O., & Altman, A. (2004). Role of plant heat-shock proteins and molecular chaperones in the abiotic stress responses. Trends in Plant Science, 9, 244–252.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work was funded by the Academy of Finland, projects 201073 (VO) and 51758 (EO). We thank Timo Oksanen for ozone fumigations.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Vladimir Ossipov.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ossipov, V., Ossipova, S., Bykov, V. et al. Application of metabolomics to genotype and phenotype discrimination of birch trees grown in a long-term open-field experiment. Metabolomics 4, 39–51 (2008). https://doi.org/10.1007/s11306-007-0097-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11306-007-0097-8

Keywords

Navigation