European Journal of Plant Pathology

, Volume 127, Issue 1, pp 21–32 | Cite as

Antifungal effect and reduction of Ulmus minor symptoms to Ophiostoma novo-ulmi by carvacrol and salicylic acid

  • Juan A. Martín
  • Alejandro SollaEmail author
  • Johanna Witzell
  • Luis Gil
  • María C. García-Vallejo


There are still no effective means to control Dutch elm disease (DED), caused by the vascular fungi Ophiostoma ulmi and O. novo-ulmi. Plant phenolics may provide a new strategy for DED control, given their known antifungal activity against pathogens and their involvement in plant defence mechanisms. The in vitro antifungal activity of salicylic acid, carvacrol, thymol, phenol, o-cresol, m-cresol, p-cresol, and 2,5-xylenol against the DED pathogens was tested. Also, the protective effect of watering Ulmus minor seedlings with these compounds was tested against O. novo-ulmi. Salicylic acid, carvacrol, and thymol showed the strongest antifungal in vitro activity, while carvacrol and salicylic acid provided the strongest in vivo protection against O. novo-ulmi (63 and 46% reduction of leaf wilting symptoms with respect to controls, respectively). The effect of the treatments on tree phenology was low, and a significant negative relation was observed between the number of days to bud burst and the leaf wilting symptoms after inoculation, probably determined by genetic differences among the elm tree progenies used. The treatments with salicylic acid, carvacrol and thymol induced the highest shift in phenolic metabolite profile with respect to control trees. The protective effect of carvacrol and salicylic acid is discussed in terms of their combined activity as antifungal compounds and as inductors of tree defence responses.


Dutch elm disease phenolic compounds tree resistance tree phenology 



Dutch elm disease


days post inoculation


salicylic acid


systemic induced resistance



The authors are very grateful to Dr. R. Raposo (INIA-CIFOR) for making suggestions for the in vitro experiment, and to M. Burón and M. A. García (Universidad Politécnica de Madrid) for their technical assistance. This work was supported by the research project INIA RTA05-151 and by an agreement established between DGMN (Ministerio de Medio Ambiente y Medio Rural y Marino) and ETSI de Montes in Madrid.


  1. Amborabé, B. E., Fleurat-Lessard, P., Chollet, J. F., & Roblin, G. (2002). Antifungal effects of salicylic acid and other benzoic acid derivatives towards Eutypa lata: structure-activity relationship. Plant Physiology and Biochemistry, 40, 1051–1060.CrossRefGoogle Scholar
  2. Brasier, C. M. (1991). Ophiostoma novo-ulmi sp-nov, causative agent of current Dutch elm disease pandemics. Mycopathologia, 115, 151–161.CrossRefGoogle Scholar
  3. Conrath, U. (2006). Systemic acquired resistance. Plant Signaling & Behavior, 1, 179–184.Google Scholar
  4. Georgiou, C. D., Tairis, N., & Sotiropoulou, A. (2000). Hydroxyl radical scavengers inhibit lateral-type sclerotial differentiation and growth in phytopathogenic fungi. Mycologia, 92, 825–834.CrossRefGoogle Scholar
  5. Heil, M., & Bostock, R. M. (2002). Induced systemic resistance (ISR) against pathogens in the context of induced plant defences. Annals of Botany, 89, 503–512.CrossRefPubMedGoogle Scholar
  6. Heimler, D., Pieroni, A., & Mittempergher, L. (1994). Plant phenolics in elms (Ulmus spp.) infected by Dutch elm disease fungus (Ophiostoma ulmi). Acta Horticulturae, 381, 638–641.Google Scholar
  7. Isman, M. B. (2000). Plant essential oils for pest and disease management. Crop Protection, 19, 603–608.CrossRefGoogle Scholar
  8. Kordali, S., Cakir, A., Ozer, H., Cakmakci, R., Kesdek, M., & Mete, E. (2008). Antifungal, phytotoxic and insecticidal properties of essential oil isolated from Turkish Origanum acutidens and its three components, carvacrol, thymol and p-cymene. Bioresource Technology, 99, 8788–8795.CrossRefPubMedGoogle Scholar
  9. Korte, F., Kvesitadze, G., Ugrekhelidze, D., Gordeziani, M., Khatisashvili, G., Buadze, O., et al. (2000). Organic Toxicants and Plants. Ecotoxicology and Environmental Safety, 47, 1–26.CrossRefPubMedGoogle Scholar
  10. Lee, S. O., Choi, G. J., Jang, K. S., Lim, H. K., Cho, K. Y., & Kim, J. C. (2007). Antifungal activity of five plant essential oils as fumigant against post-harvest and soilborne plant pathogenic fungi. Plant Pathology Journal, 23, 97–102.Google Scholar
  11. Martín, J. A., Solla, A., Woodward, S., & Gil, L. (2005). FT-IR spectroscopy as a new method for evaluating host resistance in the Dutch elm disease complex. Tree Physiology, 25, 1331–1338.PubMedGoogle Scholar
  12. Martín, J. A., Solla, A., Coimbra, M. A., Domingues, M. R., & Gil, L. (2008). Exogenous phenol increase resistance of Ulmus minor to Dutch elm disease through formation of suberin-like compounds on xylem tissues. Environmental and Experimental Botany, 64, 97–104.CrossRefGoogle Scholar
  13. Mills, P. R., & Wood, R. K. S. (1984). The effects of polyacrylic acid, acetylsalicylic acid and salicylic acid on resistance of cucumber to Colletotrichum lagenarium. Journal of Phytopathology, 111, 209–216.CrossRefGoogle Scholar
  14. Okuno, T., Nakayama, M., Okajima, N., & Furasawa, I. (1991). Systemic resistance to downy mildew and appearance of acid soluble proteins in cucumber leaves treated with biotic and abiotic inducers. Annals of the Phytopathological Society of Japan, 57, 203–211.Google Scholar
  15. Ouellette, G. B., Rioux, D., Simard, M., & Cherif, M. (2004). Ultrastructural and cytochemical studies of host and pathogens in some fungal wilt diseases: retro- and introspection towards a better understanding of DED. Investigación Agraria: Sistemas y Recursos Forestales, 13, 119–145.Google Scholar
  16. Percival, G. C. (2001). Induction of systemic acquired disease resistance in plants: potential implications for disease management in urban forestry. Journal of Arboriculture, 27, 181–192.Google Scholar
  17. Ranocha, P., Chabannes, M., Chamayou, S., Danoun, S., Jauneau, A., Boudet, A. M., et al. (2002). Laccase down-regulation causes alterations in phenolic metabolism and cell wall structure in poplar. Plant Physiology, 129, 145–155.CrossRefPubMedGoogle Scholar
  18. Raposo, R., Colgan, R., Delcan, J., & Melgarejo, P. (1995). Application of an automated quantitative method to determine fungicide resistance in Botrytis cinerea. Plant Disease, 79, 294–296.Google Scholar
  19. Roller, S., & Sheedhar, P. (2002). Carvacrol and cinnamic inhibit microbial growth in fresh cut melon and kiwifruit at 4º and 8 º C. Letters in Applied Microbiology, 35, 390–394.CrossRefPubMedGoogle Scholar
  20. Santini, A., Fagnani, A., Ferrini, F., Ghelardini, L., & Mittempergher, L. (2005). Variation among Italian and French elm clones in their response to Ophiostoma novo-ulmi inoculation. Forest Pathology, 35, 183–193.CrossRefGoogle Scholar
  21. Scheffer, R. J., Voeten, J. G. W. F., & Guries, R. P. (2008). Biological control of Dutch elm disease. Plant Disease, 92, 192–200.CrossRefGoogle Scholar
  22. Shapiro, S. S., & Wilks, M. B. (1965). An analysis of variance test for normality (complete samples). Biometrika, 52, 591–611.Google Scholar
  23. Shrivastava, V., Schinkel, H., Witzell, J., Hertzberg, M., Torp, M., Shrivastava, M. K., et al. (2007). Downregulation of high-isoelectric-point extracellular superoxide dismutase mediates alterations in the metabolism of reactive oxygen species and developmental distrurbances in hybrid aspen. Plant Journal, 49, 135–148.CrossRefGoogle Scholar
  24. Smalley, E. B., & Guries, R. P. (2000). Asian elms: sources of disease and insect resistance. In C. P. Dunn (Ed.), The elms: breeding, conservation, and disease management (pp. 215–230). Norwell: Kluwer Academic Publishers.Google Scholar
  25. Solla, A., & Gil, L. (2003). Evaluating Verticillium dahliae for biological control of Ophiostoma novo-ulmi in Ulmus minor. Plant Pathology, 52, 579–585.CrossRefGoogle Scholar
  26. Solla, A., Martín, J. A., Corral, P., & Gil, L. (2005a). Seasonal changes in wood formation of Ulmus pumila and U. minor and its relation with Dutch elm disease. New Phytologist, 166, 1025–1034.Google Scholar
  27. Solla, A., Martín, J. A., Ouellette, G., & Gil, L. (2005b). Influence of plant age on symptom development in Ulmus minor following inoculation by Ophiostoma novo-ulmi. Plant Disease, 89, 1035–1040.CrossRefGoogle Scholar
  28. Soylu, E. M., Soylu, S., & Kurt, S. (2006). Antimicrobial activities of the essential oils of various plants against tomato late blight disease agent Phytophthora infestans. Mycopathologia, 161, 119–128.CrossRefPubMedGoogle Scholar
  29. Stennes, M. A. (2000). Dutch Elm Disease Chemotherapy with Arbotect 20 S and Alamo. In C. P. Dunn (Ed.), The elms: breeding, conservation, and disease management (pp. 173–199). Norwell: Kluwer Academic Publishers.Google Scholar
  30. Tchernoff, V. (1965). Methods for screening and for the rapid selection of elms for resistance to Dutch elm disease. Acta Botanica Neerlandica, 14, 409–452.Google Scholar
  31. Webber, J. F. (2004). Experimental studies on factors influencing the transmission of Dutch elm disease. Investigación Agraria: Sistemas y Recursos Forestales, 13, 197–205.Google Scholar
  32. Witzell, J., Gref, R., & Näsholm, T. (2003). Phenolic compounds in vegetative tissues of bilberry (Vaccinium myrtillus L.). Biochemical Systematics and Ecology, 31, 115–127.CrossRefGoogle Scholar
  33. Witzell, J., & Martín, J. A. (2008). Phenolic metabolites in the resistance of northern forest trees to pathogens — past experiences and future prospects. Canadian Journal of Forest Research, 38, 2711–2727.CrossRefGoogle Scholar
  34. Zeneli, G., Krokene, P., Christiansen, E., Krekling, T., & Gershenzon, J. (2006). Methyl jasmonate treatment of mature Norway spruce (Picea abies) trees increases the accumulation of terpenoid resin components and protects against infection by Ceratocystis polonica, a bark beetle-associated fungus. Tree Physiology, 26, 977–988.PubMedGoogle Scholar

Copyright information

© KNPV 2009

Authors and Affiliations

  • Juan A. Martín
    • 1
  • Alejandro Solla
    • 2
    Email author
  • Johanna Witzell
    • 3
  • Luis Gil
    • 4
  • María C. García-Vallejo
    • 1
  1. 1.Centro de Investigación ForestalInstituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA)MadridSpain
  2. 2.Ingeniería Técnica ForestalUniversidad de ExtremaduraPlasenciaSpain
  3. 3.Southern Swedish Forest Research CentreSwedish University of Agricultural SciencesAlnarpSweden
  4. 4.Anatomía, Fisiología y Genética Forestal, ETSI de MontesUniversidad Politécnica de MadridMadridSpain

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