Regulatory gene candidates and gene expression analysis of cold acclimation in winter and spring wheat
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Freezing tolerance in plants develops through acclimation to cold by growth at low, above-freezing temperatures. Wheat is one of the most freezing-tolerant plants among major crop species and the wide range of freezing tolerance among wheat cultivars makes it an excellent model for investigation of the genetic basis of cold tolerance. Large numbers of genes are known to have altered levels of expression during the period of cold acclimation and there is keen interest in deciphering the signaling and regulatory pathways that control the changes in gene expression associated with acquired freezing tolerance. A 5740 feature cDNA amplicon microarray that was enriched for signal transduction and regulatory genes was constructed to compare changes in gene expression in a highly cold-tolerant winter wheat cultivar CDC Clair and a less tolerant spring cultivar, Quantum. Changes in gene expression over a time course of 14 days detected over 450 genes that were regulated by cold treatment and were differentially regulated between spring and winter cultivars, of these 130 are signaling or regulatory gene candidates, including: transcription factors, protein kinases, ubiquitin ligases and GTP, RNA and calcium binding proteins. Dynamic changes in transcript levels were seen at all periods of cold acclimation in both cultivars. There was an initial burst of gene activity detectable during the first day of CA, during which 90% of all genes with increases in transcript levels became clearly detectable and early expression differential between the two cultivars became more disparate with each successive period of cold acclimation.
KeywordsCold acclimation Regulatory gene Gene expression profile Wheat
We thank Matt Links, Luke McCarthy, and Bill Crosby for assistance with bioinformatics for wheat EST sequences; Olin Anderson for cDNA clones used in the microarray construction, and Ian Ferguson for advice for statistical analysis. We thank Youko Oono, Motoaki Seki and Kazuo Shinozaki for providing gene expression data for Arabidopsis. We thank the Centre for Structural Genomics, Concordia University, for assistance in the preparation and printing of the microarray.
This work was supported by a Genome Canada, Genome Prairie, and Genome Quebec grant to P.J.G., G.J.S. and F.S. and by grants from the Natural Sciences and Engineering Council of Canada to P.J.G and F.S.
Antonio F. Monroy and Ani Dryanova contributed equally to this work.
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