Planta

, Volume 226, Issue 6, pp 1459–1473

Cloning and expression analysis of candidate genes involved in wax deposition along the growing barley (Hordeum vulgare) leaf

Authors

  • Andrew Richardson
    • Division of Biological SciencesUniversity of Paisley
    • Millipore
  • Alexandre Boscari
    • Division of Biological SciencesUniversity of Paisley
    • Interactions Plantes Microorganismes et Santé VégétaleUMR INRA 1064/Université de Nice-Sophia Antipolis/CNRS 6192
  • Lukas Schreiber
    • Department of Ecophysiology, IZMBUniversity of Bonn
  • Gerhard Kerstiens
    • Department of Biological Sciences, Lancaster Environment CentreLancaster University
  • Mike Jarvis
    • Department of ChemistryUniversity of Glasgow
  • Pawel Herzyk
    • Sir Henry Wellcome Functional Genomics Facility, Institute of Biomedical and Life SciencesUniversity of Glasgow
    • Division of Biological SciencesUniversity of Paisley
    • UCD School of Biology and Environmental Science, UCD Science Centre WestUniversity College Dublin
Original Article

DOI: 10.1007/s00425-007-0585-0

Cite this article as:
Richardson, A., Boscari, A., Schreiber, L. et al. Planta (2007) 226: 1459. doi:10.1007/s00425-007-0585-0

Abstract

The aim of the present study was to isolate clones of genes which are likely to be involved in wax deposition on barley leaves. Of particular interest were those genes which encode proteins that take part in the synthesis and further modification of very long chain fatty acids (VLCFAs), the precursors of waxes. Previously, it had been shown that wax deposition commences within a spatially well-defined developmental zone along the growing barley leaf (Richardson et al. in Planta 222:472–483, 2005). In the present study, a barley microarray approach was used to screen for candidate contig-sequences (www.barleybase.org) that are expressed particularly in those leaf zones where wax deposition occurs and which are expressed specifically within the epidermis, the site of wax synthesis. Candidate contigs were used to screen an established in-house cDNA library of barley. Six full-length coding sequences clones were isolated. Based on sequence homologies, three clones were related to Arabidopsis CER6/CUT1, and these clones were termed HvCUT1;1, HvCUT1;2 and HvCUT1;3. A fourth clone, which was related to Arabidopsis Fiddlehead (FDH), was termed HvFDH1;1. These clones are likely to be involved in synthesis of VLCFAs. A fifth and sixth clone were related to Arabidopsis CER1, and were termed HvCER1;1 and HvCER1;2. These clones are likely to be involved in the decarbonylation pathway of VLCFAs. Semi-quantitative RT-PCR confirmed microarray expression data. In addition, expression analyses at 10-mm resolution along the blade suggest that HvCUT1;1 (and possibly HvCUT1;2) and HvCER1;1 are involved in commencement of wax deposition during barley leaf epidermal cell development.

Keywords

CuticleEpidermal cell developmentHordeumLeaf growthMicroarrayVery long chain fatty acid elongationWax

Abbreviations

ACP

Acyl-carrier protein

FAE

Fatty acid elongase

FAS

Fatty acid synthase

GL1

Glossy 1

GL8

Glossy 8

KCR

3(β)-ketoacyl-reductase

KCS

3(β)-Ketoacyl-CoA synthase

LACS

Long-chain acyl-CoA synthetase

POE

Point of emergence

POLI

Point of leaf insertion

VLCFA

Very long chain fatty acid

Supplementary material

425_2007_585_MOESM1_ESM.xls (2.7 mb)
(XLS 2723 kb)

Copyright information

© Springer-Verlag 2007