Expression profiles of a set of Cor/Lea genes were assessed during early stages of cold acclimation in seedlings of two wheat cultivars, which showed contrasting levels of freezing tolerance. These Cor/Lea family members consisted of three EST clones and 13 previously identified cDNA clones of wheat and rye. Northern blot analysis using RNA extracted from seedling leaves and roots showed that most of the genes exhibited a quite similar time-course of expression, although with different expression levels: They rapidly responded to low temperature and their transcript levels reached high plateaus within 3-5 days. The overall gene expression profiles were correlated with the time-dependent development and the level of freezing tolerance under low temperature in the two cultivars. Western blot analysis of protein accumulation further verified this observation. Abscissic acid response was proved for at least four genes. Light was stimulatory to most of the genes, and this positive light response associated with low temperature occurred not only in leaf-specific genes but also in leaf/root-expressed genes. Taken together, the present results suggest that the Cor/Lea gene family represents a major group of downstream genes involved in the ABA-dependent and -independent signal pathways and that most of them are co-regulated in determining freezing tolerance in wheat seedlings.

Wcor15, a member of the wheat cold-responsive (Cor) gene family, has been isolated and characterized. The deduced polypeptide WCOR15 (MW=14.7 kDa) showed high homology to the previously identified wheat and barley COR proteins. Southern blot analysis using diploid, tetraploid and hexaploid wheat and diploid Aegilops species showed that the wheat and related wild genomes possessed multiple copies of Wcor15 homologues. Five copies were assigned to the homologous group 2 chromosomes by nulli-tetrasomic analysis. Northern blot analysis showed that expression of Wcor15 was specifically induced by low-temperature. Homologous transcripts accumulated in leaves, and light markedly increased their steady-state levels. Bombardment-mediated transient expression analysis of a chimeric CaMV 35S::Wcor15-GFP construct showed protein-targeting to epidermal guard cell chloroplasts in excised spiderwort leaves. A promoter of Wcor15 contained at least three CRT/DRE-like sequence motifs found in Arabidopsis Cor genes and induced the reporter GUS gene expression in leaves of transgenic tobacco plants under low-temperature and light conditions. These results suggest that the functional Cor gene system involving the CRT/DRE cis-element is conserved in both monocotyledonous and dicotyledonous plants.