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ABA-responsive element binding protein (AREB) and ABA-responsive element binding factor (ABF), members of the basic region/leucine zipper (bZIP)-type protein family, act as major transcription factors in ABA-responsive gene expression under abiotic stress conditions in Arabidopsis. Barley HvABI5 and rice transcription factor responsible for ABA regulation 1 (TRAB1) are homologues of AREB/ABF and are expressed in drought- and ABA-treated seedlings. However, no direct evidence has shown an association of an AREB/ABF-type transcription factor with stress tolerance in cereals. To understand the molecular basis of abiotic stress tolerance through a cereal AREB/ABF-type transcription factor, a wheat HvABI5 ortholog, Wabi5, was isolated and characterized. Wabi5 expression was activated by low temperature, drought and exogenous ABA treatment, and its expression pattern differed between two wheat accessions with distinct levels of stress tolerance and ABA sensitivity. Wabi5-expressing transgenic tobacco plants showed a significant increase in tolerance to abiotic stresses such as freezing, osmotic and salt stresses and a hypersensitivity to exogenous ABA in the seedling stage compared with wild-type plants. Expression of a GUS reporter gene under the control of promoters of three wheat cold-responsive/late embryogenesis abundant (Cor/Lea) genes, Wdhn13, Wrab18 and Wrab19, was enhanced by ectopic Wabi5 expression in wheat callus and tobacco plants. These results clearly indicated that WABI5 functions as a transcriptional regulator of the Cor/Lea genes in multiple abiotic stress responses in common wheat.

Cereal lip19 genes encoding bZIP-type transcription factors are assumed to play a regulatory role in gene expression during the cold acclimation process. However, no direct evidence shows an association of LIP19-type bZIPs with stress tolerance or activation of stress-responsive Cor/Lea genes. To understand the molecular basis of development of abiotic stress tolerance through the LIP19 transcription factor, a wheat lip19 homologue, Wlip19, was isolated and characterized. Wlip19 expression was activated by low temperature in seedlings and was higher in a freezing-tolerant cultivar than in a freezing-sensitive one. Wlip19 also responded to drought and exogenous ABA treatment. Wlip19-expressing transgenic tobacco showed a significant increase in abiotic stress tolerance, especially freezing tolerance. Expression of a GUS reporter gene under the control of promoter sequences of four wheat Cor/Lea genes, Wdhn13, Wrab17, Wrab18, and Wrab19, was enhanced by Wlip19 expression in wheat callus and tobacco plants. These results indicate that WLIP19 acts as a transcriptional regulator of Cor/Lea genes in the development of abiotic stress tolerance. Moreover, direct protein-protein interaction between WLIP19 and a wheat OBF1 homologue TaOBF1, another bZIP-type transcription factor, was observed, suggesting that this interaction is conserved in cereals.