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JXB Advance Access published online on May 7, 2004
Journal of Experimental Botany, doi:10.1093/jxb/erh153
© 2004 by Oxford University Press
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Received January 14, 2004; accepted March 15, 2004
© 2004 Society for Experimental Biology

RESEARCH PAPER

Expression analysis of a chicory fructan 1-exohydrolase gene reveals complex regulation by cold

An Michiels 1*, Andre Van Laere 1, Wim Van den Ende 1, and Mark Tucker 2

1 Laboratory for Molecular Plant Physiology, Institute of Botany and Microbiology, K.U. Leuven, Kasteelpark Arenberg 31, B-3001 Leuven, Belgium
2 United States Department of Agriculture, Agricultural Research Service, Soybean Genomics and Improvement Laboratory, Building 6, BARC-West, 10300 Baltimore Ave., Beltsville, MD 20705-2350, USA

* To whom correspondence should be addressed. E-mail: michiels{at}wam.umd.edu.


  Abstract

The gene for a recently identified cDNA, 1-FEH IIa, encoding a fructan 1-exohydrolase was isolated and cloned from Cichorium intybus and a 1149 bp promoter fragment was characterized. An analysis of the genomic 1-FEH IIa sequence indicated that the gene (FEHIIa) consists of six introns and seven exons, which is similar to plant invertase genes. Like invertase genes, FEHIIa also contains the 9 nt mini-exon encoding the tripeptide DPN. A database search for cis-acting response elements within its promoter identified multiple elements that appear to have relevance to cold-induced expression of the gene in field-grown roots. Promoter analysis by transient expression assay demonstrated that the FEHIIa gene promoter is highly expressed in etiolated Cichorium leaves and cold-stored roots, which correlated well with the high level expression detected by RNA blot analysis. Cold also enhanced FEHIIa reporter gene expression in green leaves, however, the reporter gene activity was much lower compared with similar induction experiments in etiolated leaves. Promoter deletion analysis demonstrated the presence of potential cold-responsive ABRE and/or CRT/DRE elements in the -22 to -172 region, while regions -933 to -717 and -493 to -278 contain elements that can down-regulate expression at the conditions used. Characterization of the FEHIIa promoter may provide tools to study cold-induced expression and to increase freezing tolerance in agricultural crops.

Key words: Cold induction, fructan, fructan exohydrolase, gene structure, promoter analysis.


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