Skip to main content
SpringerLink
Log in

Flavonoid biosynthesis and degradation play a role in early defence responses of bilberry (Vaccinium myrtillus) against biotic stress

  • Original Research
  • Published:
European Journal of Plant Pathology Aims and scope Submit manuscript

Abstract

Bilberry (Vaccinium myrtillus) represents one of the richest flavonoid sources among plants. Flavonoids play variable, species-dependent roles in plant defences. In bilberry, flavonoid metabolism is activated in response to solar radiation but not against mechanical injury. In this paper, the defence reaction and biosynthesis of phenolic compounds of bilberry was studied after infection by a fungal endophyte (Paraphaeosphaeria sp.) and a pathogen (Botrytis cinerea). The defence response of bilberry was faster against the endophyte than the pathogen. All flavonoid biosynthesis genes tested were activated by each infection. Biosynthesis and accumulation of phenolic acids, flavan-3-ols and oligomeric proanthocyanidins were clearly elevated in both infected samples. Infection by the pathogen promoted specifically accumulation of epigallocatechin, quercetin-3-glucoside, quercetin-3-O-α-rhamnoside, quercetin-3-O-(4”-HMG)-R-rhamnoside, chlorogenic acid and coumaroyl quinic acid. The endophyte-infected plants had a higher content of quercetin-3-glucuronide and coumaroyl iridoid. Therefore, accumulation of individual phenolic compounds could be specific for each infection. Quantity of insoluble proanthocyanidins was the highest in control plants, suggesting that they might act as storage compounds and become activated by degradation upon infection.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

Abbreviations

CHS:

chalcone synthase

DFR:

dihydroflavonol 4-reductase

ANS:

anthocyanidin synthase

ANR:

anthocyanidin reductase

PR4:

pathogenesis-related protein 4

MEA:

malt-extract agar

References

  • Carlsen, S. C. K., Understrup, A., Fomsgaard, I. S., Mortensen, A. G., & Ravnskov, S. (2008). Flavonoids in roots of white clover: interaction of arbuscular mycorrhizal fungi and a pathogenic fungus. Plant Soil, 302, 33–43.

    Article  CAS  Google Scholar 

  • Chenna, R., Sugawara, H., Koike, T., Lopez, R., Gibson, T. J., Higgins, D. G., et al. (2003). Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Research, 31, 497–500.

    Article  Google Scholar 

  • de Colmenares, N. G., Ramirez-Martinez, J. R., Aldana, J. O., Ramos-Nino, M. E., Clifford, M. N., Pekerar, S., et al. (1998). Isolation, characterisation and determination of biological activity of coffee proanthocyanidins. Journal of the Science of Food and Agriculture, 77, 368–372.

    Article  Google Scholar 

  • Dixon, R. A., & Paiva, N. L. (1995). Stress-Induced Phenylpropanoid Metabolism. Plant Cell, 7, 1085–1097.

    Article  CAS  PubMed  Google Scholar 

  • Dixon, R. A., Achnine, L., Kota, P., Liu, C.-J., Reddy, M. S. S., & Wang, L. (2002). The phenylpropanoid pathway and plant defence—a genomics perspective. Molecular Plant Pathology, 3, 371–390.

    Article  CAS  Google Scholar 

  • Felsenstein, J. (1989). PHYLIP — Phylogeny Inference Package (Version 3.2). Cladistics, 5, 164–166.

    Google Scholar 

  • Fukuhara, M. (2002). Three Phaeosphaeria species and Paraphaeosphaeria michotii isolated from Phragmites leaves in Osaka, Japan. Mycoscience, 43, 375–382.

    Article  Google Scholar 

  • Ganley, R. J., & Newcombe, G. (2006). Fungal endophytes in seeds and needles of Pinus monticola. Mycological Research, 110, 318–327.

    Article  PubMed  Google Scholar 

  • Goetz, G., Fkyerat, A., Metais, N., Kunz, M., Tabacchi, R., Pezet, R., et al. (1999). Resistance factors to grey mould in grape berries: identification of some phenolics inhibitors of Botrytis cinerea stilbene oxidase. Phytochemistry, 52, 759–767.

    Article  CAS  Google Scholar 

  • Hébert, C., Charles, M. T., Gauthier, L., Willemot, C., Khanizadeh, S., & Cousineau, J. (2002). Strawberry proanthocyanidins: Biochemical markers for Botrytis cinerea resistance and shelf-life predictability. Acta Horticulturae, 567, 659–662.

    Google Scholar 

  • Hildebrandt, P. D., McRae, K. B., & Lu, X. (2001). Factors affecting flower infection and disease severity of lowbush blueberry by Botrytis cinerea. Canadian Journal of Plant Pathology, 23, 364–370.

    Google Scholar 

  • Jaakola, L., Tolvanen, A., Laine, K., & Hohtola, A. (2001a). Effect of N6-isopentenyladenine concentration on growth initiation in vitro and rooting of bilberry and lingonberry microshoots. Plant Cell Tissue and Organ Culture, 66, 73–77.

    Article  CAS  Google Scholar 

  • Jaakola, L., Pirttilä, A. M., & Hohtola, A. (2001b). Isolation of RNA from bilberry (Vaccinium myrtillus). Molecular Biotechnology, 19, 201–203.

    Article  CAS  Google Scholar 

  • Jaakola, L., Määttä-Riihinen, K., Kärenlampi, S., & Hohtola, A. (2004). Activation of flavonoid biosynthesis by solar radiation in bilberry (Vaccinium myrtillus) leaves. Planta, 218, 721–728.

    Article  CAS  PubMed  Google Scholar 

  • Jaakola, L., Koskimäki, J. J., Riihinen, K., Tolvanen, A., & Hohtola, A. (2008). Effect of wounding on chalcone synthase and pathogenesis related PR10 gene expression and content of phenolic compounds in bilberry leaves. Biologia Plantarum, 52, 391–395.

    Article  CAS  Google Scholar 

  • Larose, G., Chenevert, R., Moutoglis, P., Gagne, S., Piche, Y., & Vierheilig, H. (2002). Flavonoid levels in roots of Medicago sativa are modulated by the developmental stage of the symbiosis and the root colonizing arbuscular mycorrhizal fungus. Journal of Plant Physiology, 159, 1329–1339.

    Article  CAS  Google Scholar 

  • Laukkanen, H., Soini, H., Kontunen-Soppela, S., Hohtola, A., & Viljanen, M. (2000). A mycobacterium isolated from tissue cultures of mature Pinus sylvestris interferes with growth of Scots pine seedlings. Tree Physiology, 20, 915–920.

    PubMed  Google Scholar 

  • Lee, M. H., & Bostock, R. M. (2007). Fruit exocarp phenols in relation to quiescence and development of Monilinia fructicola infections in Prunus spp.: A role for cellular redox? Phytopathology, 97, 269–277.

    Article  CAS  PubMed  Google Scholar 

  • Logemann, E., & Hahlbrock, K. (2002). Crosstalk among stress responses in plants: Pathogen defence overrides UV protection through an inversely regulated ACE/ACE type of light-responsive gene promoter unit. Proceedings of the National Academy of Sciences, 99, 2428–2432.

    Article  CAS  Google Scholar 

  • Määttä, K., Kamal-Eldin, A., & Törrönen, R. (2001). Phenolic compounds in berries of black, red, green, and white currants (Ribes sp.). Antioxidants & Redox Signaling, 3, 981–993.

    Article  Google Scholar 

  • McKhann, H. I., Paiva, N. L., Dixon, R. A., & Hirsch, A. M. (1997). Chalcone synthase transcripts are detected in alfalfa root hairs following inoculation with wild-type Rhizobium meliloti. Molecular Plant-Microbe Interactions, 10, 50–58.

    Article  CAS  Google Scholar 

  • Miranda, M., Ralph, S. G., Mellway, R., White, R., Heath, M. C., Bohlmann, J., et al. (2007). The transcriptional response of hybrid poplar (Populus trichocarpa x P-deltoides) to infection by Melampsora medusae leaf rust involves induction of flavonoid pathway genes leading to the accumulation of proanthocyanidins. Molecular Plant-Microbe Interactions, 20, 816–831.

    Article  CAS  PubMed  Google Scholar 

  • Mohr, U., Lange, J., Boller, T., Wiemken, A., & Vogeli-Lange, R. (1998). Plant defence genes are induced in the pathogenic interaction between bean roots and Fusarium solani, but not in the symbiotic interaction with the arbuscular mycorrhizal fungus Glomus mosseae. New Phytologist, 138, 589–598.

    Article  CAS  Google Scholar 

  • Morazzoni, P., & Bombardelli, E. (1996). Vaccinium myrtillus L. Fitoterapia, 67, 3–29.

    CAS  Google Scholar 

  • Muthuswamy, S., & Vasantha, R. H. P. (2007). Fruit phenolics as natural antimicrobial agents : Selective antimicrobial activity of catechin, chlorogenic acid and phloridzin. International Journal of Food, Agriculture and Environment, 5, 81–85.

    CAS  Google Scholar 

  • Pandey, M. K., Sarma, B. K., Singh, D. P., & Singh, U. P. (2007). Biochemical investigations of sclerotial exudates of Sclerotium rolfsii and their antifungal activity. Journal of Phytopathology, 155, 84–89.

    Article  CAS  Google Scholar 

  • Pehkonen, T., Koskimäki, J. J., Riihinen, K., Pirttilä, A. M., Hohtola, A., Jaakola, L., et al. (2008). Artificial infection of Vaccinium vitis-idaea L. and defence responses to Exobasidium species. Physiological and Molecular Plant Pathology, 12, 146–150.

    Google Scholar 

  • Pirttilä, A. M., Kämäräinen, T., Hirsikorpi, M., Jaakola, L., & Hohtola, A. (2001). DNA isolation methods for medicinal and aromatic plants. Plant Molecular Biology Reporter, 19, 273a–f.

    Article  Google Scholar 

  • Polya, G. M., & Foo, L. Y. (1994). Inhibition of eukaryote signal-regulated protein kinases by plant-derived catechin-related compounds. Phytochemistry, 35, 1399–1405.

    Article  CAS  PubMed  Google Scholar 

  • Pozo, M. J., Azcón-Aguilar, C., Dumas-Gaudot, E., & Barea, J. M. (1999). β-1, 3-Glucanase activities in tomato roots inoculated with arbuscular mycorrhizal fungi and/or Phytophthora parasitica and their possible involvement in bioprotection. Plant Science, 141, 149–157.

    Article  CAS  Google Scholar 

  • Punyasiri, P. A. N., Tanner, G. J., Abeysinghe, S. B., Kumar, V., Campbell, P. M., & Pradeepa, N. H. L. (2004). Exobasidium vexans infection of Camellia sinensis increased 2, 3-cis isomerization and gallate esterification of proanthocyanidins. Phytochemistry, 65, 2987–2994.

    Article  CAS  Google Scholar 

  • Salzer, P., Feddermann, N., Wiemken, A., Boller, T., & Staehelin, C. (2004). Sinorhizobium meliloti-induced chitinase gene expression in Medicago truncatula ecotype R108-1: a comparison between symbiosis-specific class V and defence-related class IV chitinases. Planta, 219, 626–638.

    Article  CAS  PubMed  Google Scholar 

  • Sauer, M., Lu, P., Sangar, R., Kennedy, S., Polishook, J., Bills, G., et al. (2002). Estimating polyketide metaholic potential among non-sporulating fungal endophytes of Vaccinium macrocarpon. Mycological Research, 106, 460–470.

    Article  CAS  Google Scholar 

  • Schulz, B., Römmert, A.-K., Dammann, U., Aust, H.-J., & Strack, D. (1999). The endophyte-host interaction: a balanced antagonism? Mycological Research, 103, 1275–1283.

    Article  Google Scholar 

  • Tolonen, A., & Uusitalo, J. (2004). Fast screening method for the analysis of total flavonoid content in plants and foodstuffs by high-performance liquid chromatography/electrospray ionization time-of-flight mass spectrometry with polarity switching. Rapid Communications in Mass Spectrometry, 18, 3113–3122.

    Article  CAS  PubMed  Google Scholar 

  • Treutter, D. (2005). Significance of flavonoids in plant resistance and enhancement of their biosynthesis. Plant Biology, 7, 581–591.

    Article  CAS  PubMed  Google Scholar 

  • Veluri, R., Weir, T. L., Bais, H. P., Termitz, F. R. S., & Ivanco, J. M. V. (2004). Phytotoxic and antimicrobial activities of catechin derivatives. Journal of Agricultural and Food Chemistry, 52, 1077–1082.

    Article  CAS  PubMed  Google Scholar 

  • Vlassova, T., Likhachev, A. & Blintsov, A. (2000). Accumulation of abscisic acid in culture filtrates of selected species of genus Botrytis. (Paper presented at the XIIth International Botrytis symposium (p. 23) Reims, France)

  • White, T. J., Burns, T., Lee, S., & Taylor, J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In M. A. Innis, D. H. Gelfand, J. J. Sninsky & T. J. White (Eds.), PCR Protocols (pp. 315–322). San Diego: Academic.

    Google Scholar 

  • Witzell, J., & Shevtsova, A. (2004). Nitrogen-induced changes in phenolics of Vaccinium myrtillus - implications for interaction with a parasitic fungus. Journal of Chemical Ecology, 30, 1937–1956.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Dr. P. J. Fisher (University of Portsmouth, Portsmouth, UK) for the advice on endophyte isolation from the Ericaceae. This work was supported by the Ella and Georg Ehrnrooth Foundation and Academy of Finland (No. 118569), and is part of the Endis Network (Discovery and Development of Antibacterials from Endophytes) at the University of Oulu.

Author information

Authors and Affiliations

  1. Department of Biology, University of Oulu, PO Box 3000, Oulu, 90014, Finland

    Janne J. Koskimäki, Laura Jaakola, Marja Suorsa, Anna Maria Pirttilä & Anja Hohtola

  2. Department of Chemistry, University of Oulu, PO Box 3000, Oulu, 90014, Finland

    Juho Hokkanen & Sampo Mattila

  3. Novamass Ltd, Oulu, Finland

    Juho Hokkanen & Ari Tolonen

Authors
  1. Janne J. Koskimäki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Juho Hokkanen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Laura Jaakola
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Marja Suorsa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ari Tolonen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Sampo Mattila
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Anna Maria Pirttilä
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Anja Hohtola
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Anna Maria Pirttilä.

Additional information

Janne J. Koskimäki and Juho Hokkanen have equally contributed to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Koskimäki, J.J., Hokkanen, J., Jaakola, L. et al. Flavonoid biosynthesis and degradation play a role in early defence responses of bilberry (Vaccinium myrtillus) against biotic stress. Eur J Plant Pathol 125, 629–640 (2009). https://doi.org/10.1007/s10658-009-9511-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10658-009-9511-6

Keywords

Search

Navigation