European Journal of Forest Research

, Volume 133, Issue 4, pp 573–583 | Cite as

Low-density Ceratocystis polonica inoculation of Norway spruce (Picea abies) triggers accumulation of monoterpenes with antifungal properties

  • Metka Novak
  • Andreja Urbanek Krajnc
  • Ljerka Lah
  • Neja Zupanec
  • Nada Kraševec
  • Mitja Križman
  • Joerg Bohlmann
  • Radovan Komel
Original Paper


Among the most devastating pests of Norway spruce (Picea abies) are the European spruce bark beetle (Ips typographus) and the associated pathogenic blue-stain fungus Ceratocystis polonica. Following attack and colonization, the beetle and the fungus must cope with induced host chemical defenses, such as monoterpenes that are generally thought to be toxic to both symbionts. The goal of this study was to better understand the response of Norway spruce following C. polonica inoculation at low density that does not overwhelm the tree and to identify monoterpenes mobilized toward the fungus. We inoculated healthy mature trees and monitored monoterpene profiles 2, 3, and 5 months post-inoculation. We also exposed three different C. polonica strains to the most abundant or significantly up-regulated monoterpenes to determine differences in monoterpene toxicity and resistance among strains. Total monoterpene levels, including limonene, were increased at 2 and 3 months after inoculation and had dropped after 5 months. In in vitro assays, all monoterpenes were inhibitory to C. polonica. Limonene and β-pinene were the most potent inhibitors of fungal growth. The extent of inhibition varied between the three strains tested. These results showed a defense response of Norway spruce to C. polonica, in which limonene may play a critical role in inhibiting the spread of the fungus. We also showed that differences between strains of C. polonica must be taken into account when assessing the role of the fungus in this bark beetle–symbiont system.


Blue-stain fungi Monoterpene Growth inhibition Limonene 



We thank Prof. Dr. Mario Lešnik from Faculty of Agriculture and Life Sciences, Peter Kramer from Maribor University Agriculture Centre, and Ciril Zupan from the Slovenian Forest Service for permission to use the experimental field in Pivola, Hoče, and support during the experimental work. We gratefully acknowledge support from Alfonso Lara Quesada for the assistance with data analysis and Justin G. A. Whitehill for help with the statistical analysis. We thank Mateja Felicijan for helping with the field sample collection and technical assistance with sample preparations. We thank Janez Gorenšek, Andreja Šober, Maja Svenšek, Peter Drevenšek, and Robi Gjergjek for excellent assistance in the field.

Supplementary material

10342_2013_772_MOESM1_ESM.pdf (509 kb)
Supplementary material 1 (PDF 509 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Metka Novak
    • 1
  • Andreja Urbanek Krajnc
    • 2
  • Ljerka Lah
    • 1
  • Neja Zupanec
    • 1
  • Nada Kraševec
    • 1
  • Mitja Križman
    • 1
  • Joerg Bohlmann
    • 4
  • Radovan Komel
    • 1
    • 3
  1. 1.National Institute of ChemistryLjubljanaSlovenia
  2. 2.Faculty of Agriculture and Life SciencesUniversity of MariborHočeSlovenia
  3. 3.Faculty of Medicine, Institute of BiochemistryUniversity of LjubljanaLjubljanaSlovenia
  4. 4.Michael Smith LaboratoriesUniversity of British ColumbiaVancouverCanada

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