, 224:865 | Cite as

Ethylene in induced conifer defense: cDNA cloning, protein expression, and cellular and subcellular localization of 1-aminocyclopropane-1-carboxylate oxidase in resin duct and phenolic parenchyma cells

  • J. W. Hudgins
  • Steven G. Ralph
  • Vincent R. Franceschi
  • Jörg BohlmannEmail author
Original Article


Members of the Pinaceae family have complex chemical defense strategies. Conifer defenses associated with specialized cell types of the bark involve constitutive and inducible accumulation of phenolic compounds in polyphenolic phloem parenchyma cells and oleoresin terpenoids in resin ducts. These defenses can protect trees against insect herbivory and fungal colonization. The phytohormone ethylene has been shown to induce the same anatomical and cellular defense responses that occur following insect feeding, mechanical wounding, or fungal inoculation in Douglas fir (Pseudotsuga menziesii) stems (Hudgins and Franceschi in Plant Physiol 135:2134–2149, 2004). However, very little is known about the genes involved in ethylene formation in conifer defense or about the temporal and spatial patterns of their protein expression. The enzyme 1-aminocyclopropane-1-carboxylate oxidase (ACO) catalyzes the final step in ethylene biosynthesis. We cloned full-length and near full-length ACO cDNAs from three conifer species, Sitka spruce (Picea sitchensis), white spruce (P. glauca), and Douglas fir, each with high similarity to Arabidopsis thaliana ACO proteins. Using an Arabidopsis anti-ACO antibody we determined that ACO is constitutively expressed in Douglas fir stem tissues and is up-regulated by mechanical wounding, consistent with the wound-induced increase of ethylene levels. Immunolocalization showed cytosolic ACO is predominantly present in specialized cell types of the wound-induced bark, specifically in epithelial cells of terpenoid-producing cortical resin ducts, in polyphenolic phloem parenchyma cells, and in ray parenchyma cells.


ACC oxidase Douglas fir Hormone Immunolocalization Insect Picea 



Confocal and TEM microscopy was done in the Electron Microscopy Center, Washington State University. We thank Sharon Jancsik for technical assistance with cDNA cloning and RNA isolation. This work was generously supported by grants to JB from Genome Canada, Genome British Columbia, and the province of British Columbia; the Natural Sciences and Engineering Research Council of Canada (NSERC); and with infrastructure funds from the Canadian Foundation for Innovation (CFI) and the British Columbia Knowledge and Development Funds (BCKDF). This paper is dedicated in memory of Dr. Vincent R. Franceschi (1953–2005). Vince was a leading scientist who contributed greatly to plant sciences through his teaching, research, generous collaborative support, and countless other activities. His work on the cell biology of conifer defense has been a great inspiration and provided an important foundation for many current and future studies.


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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • J. W. Hudgins
    • 1
    • 4
  • Steven G. Ralph
    • 1
  • Vincent R. Franceschi
    • 4
  • Jörg Bohlmann
    • 1
    • 2
    • 3
    Email author
  1. 1.Michael Smith LaboratoriesUniversity of British ColumbiaVancouverCanada
  2. 2.Department of BotanyUniversity of British ColumbiaVancouverCanada
  3. 3.Department of Forest SciencesUniversity of British ColumbiaVancouverCanada
  4. 4.School of Biological SciencesWashington State UniversityPullmanUSA

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