Plant Molecular Biology

, Volume 65, Issue 3, pp 311–328

Expression profiling of the lignin biosynthetic pathway in Norway spruce using EST sequencing and real-time RT-PCR

  • Sanna Koutaniemi
  • Tino Warinowski
  • Anna Kärkönen
  • Edward Alatalo
  • Carl G. Fossdal
  • Pekka Saranpää
  • Tapio Laakso
  • Kurt V. Fagerstedt
  • Liisa K. Simola
  • Lars Paulin
  • Stephen Rudd
  • Teemu H. Teeri
Article

DOI: 10.1007/s11103-007-9220-5

Cite this article as:
Koutaniemi, S., Warinowski, T., Kärkönen, A. et al. Plant Mol Biol (2007) 65: 311. doi:10.1007/s11103-007-9220-5

Abstract

Lignin biosynthesis is a major carbon sink in gymnosperms and woody angiosperms. Many of the enzymes involved are encoded for by several genes, some of which are also related to the biosynthesis of other phenylpropanoids. In this study, we aimed at the identification of those gene family members that are responsible for developmental lignification in Norway spruce (Picea abies (L.) Karst.). Gene expression across the whole lignin biosynthetic pathway was profiled using EST sequencing and quantitative real-time RT-PCR. Stress-induced lignification during bending stress and Heterobasidion annosum infection was also studied. Altogether 7,189 ESTs were sequenced from a lignin forming tissue culture and developing xylem of spruce, and clustered into 3,831 unigenes. Several paralogous genes were found for both monolignol biosynthetic and polymerisation-related enzymes. Real-time RT-PCR results highlighted the set of monolignol biosynthetic genes that are likely to be responsible for developmental lignification in Norway spruce. Potential genes for monolignol polymerisation were also identified. In compression wood, mostly the same monolignol biosynthetic gene set was expressed, but peroxidase expression differed from the vertically grown control. Pathogen infection in phloem resulted in a general up-regulation of the monolignol biosynthetic pathway, and in an induction of a few new gene family members. Based on the up-regulation under both pathogen attack and in compression wood, PaPAL2, PaPX2 and PaPX3 appeared to have a general stress-induced function.

Keywords

Compression woodESTHeterobasidion annosumLignin biosynthesisPicea abiesReal-time RT-PCR

Abbreviations

CAD

Cinnamyl alcohol dehydrogenase

CADL

CAD-like

CCOMT

Caffeoyl-CoA O-methyltransferase

CCR

Cinnamoyl-CoA reductase

COMT

Caffeic acid O-methyltransferase

CoA

Coenzyme A

Ct

Cycle threshold

C3H

p-coumarate-3-hydroxylase

C4H

Cinnamate-4-hydroxylase

EST

Expressed sequence tag

F5H/CAld5H

Ferulate/coniferaldehyde-5-hydroxylase

G

Guaiacyl lignin

H

Hydroxyphenyl lignin

HCT

Hydroxycinnamoyl:CoA shikimate/quinate hydroxycinnamoyl transferase

PAL

Phenylalanine ammonia-lyase

RT-PCR

Reverse transcription polymerase chain reaction

RSCL

Released suspension culture lignin

S

Syringyl lignin

SAD

Sinapaldehyde dehydrogenase

4CL

4-coumarate:CoA ligase

Supplementary material

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Sanna Koutaniemi
    • 1
  • Tino Warinowski
    • 1
  • Anna Kärkönen
    • 1
  • Edward Alatalo
    • 2
  • Carl G. Fossdal
    • 3
  • Pekka Saranpää
    • 4
  • Tapio Laakso
    • 4
  • Kurt V. Fagerstedt
    • 5
  • Liisa K. Simola
    • 5
  • Lars Paulin
    • 2
  • Stephen Rudd
    • 6
  • Teemu H. Teeri
    • 1
  1. 1.Department of Applied BiologyUniversity of HelsinkiHelsinkiFinland
  2. 2.Institute of BiotechnologyUniversity of HelsinkiHelsinkiFinland
  3. 3.Norwegian Forest and Landscape InstituteÅsNorway
  4. 4.Finnish Forest Research Institute MetlaVantaaFinland
  5. 5.Department of Biological and Environmental SciencesUniversity of HelsinkiHelsinkiFinland
  6. 6.Centre for BiotechnologyTurkuFinland