Plant Molecular Biology

, Volume 44, Issue 6, pp 733–745

Molecular cloning and functional expression of a stress-induced multifunctional O-methyltransferase with pinosylvin methyltransferase activity from Scots pine (Pinus sylvestris L.)

  • Hélène Chiron
  • Alain Drouet
  • Anne-Catherine Claudot
  • Christoph Eckerskorn
  • Monika Trost
  • Werner Heller
  • Dieter Ernst
  • Heinrich SandermannJr
Article

Abstract

Formation of pinosylvin (PS) and pinosylvin 3-O-monomethyl ether (PSM), as well as the activities of stilbene synthase (STS) and S-adenosyl-l-methionine (SAM):pinosylvin O-methyltransferase (PMT), were induced strongly in needles of Scots pine seedlings upon ozone treatment, as well as in cell suspension cultures of Scots pine upon fungal elicitation. A SAM-dependent PMT protein was purified and partially characterised. A cDNA encoding PMT was isolated from an ozone-induced Scots pine cDNA library. Southern blot analysis of the genomic DNA suggested the presence of a gene family. The deduced protein sequence showed the typical highly conserved regions of O-methyltransferases (OMTs), and average identities of 20–56% to known OMTs. PMT expressed in Escherichia coli corresponded to that of purified PMT (40 kDa) from pine cell cultures. The recombinant enzyme catalysed the methylation of PS, caffeic acid, caffeoyl-CoA and quercetin. Several other substances, such as astringenin, resveratrol, 5-OH-ferulic acid, catechol and luteolin, were also methylated. Recombinant PMT thus had a relatively broad substrate specificity. Treatment of 7-year old Scots pine trees with ozone markedly increased the PMT mRNA level. Our results show that PMT represents a new SAM-dependent OMT for the methylation of stress-induced pinosylvin in Scots pine needles.

cDNA cloning elicitor ozone pinosylvin O-methyltransferase Pinus sylvestris stilbene 

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Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Hélène Chiron
    • 1
    • 2
  • Alain Drouet
    • 2
  • Anne-Catherine Claudot
    • 2
  • Christoph Eckerskorn
    • 3
  • Monika Trost
    • 1
  • Werner Heller
    • 1
  • Dieter Ernst
    • 1
  • Heinrich SandermannJr
    • 1
  1. 1.Institute of Biochemical Plant PathologyGSF-National Research Center for Environment and HealthNeuherbergGermany
  2. 2.Biology DepartmentUniversity of OrléansOrléans Cedex 2France
  3. 3.MPI für BiochemieMartinsriedGermany

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