The effects of the ectopic expression of a barley transcription factor (HvCBF4) under the control of a constitutive (maize Ubi1) or a stress-inducible (Arabidopsis RD29A) promoter in the abiotic stress response in rice (Oryza sativa L.) was investigated. The transformed plants were analyzed both at molecular and physiological level and the AtRD29A::HvCBF4 plants were further analyzed using the GeneChipA (R) rice genome array under control conditions. Only the plants constitutively expressing HvCBF4 have shown increased survival to drought stress, but not to cold or high-salinity. These plants have also shown better photosynthetic capacity, as determined by chlorophyll fluorescence. Plants expressing AtRD29A::HvCBF4 did not show increased survival to any of the stresses applied. However in the GeneChipA (R) microarray, these plants have shown up-regulation of many stress-responsive genes (> 400) as compared to non-transformed plants. Interestingly, RT-PCR analysis revealed not only differential gene expression between roots and shoots, but also between transgenic lines with the different promoters. Our results indicate that different HvCBF4 expression levels resulted in different transcriptomes and drought tolerance. Given that AtRD29A::HvCBF4 plants did not show increased tolerance to any of the imposed stresses, we may conclude that this promoter may be inappropriate for rice transformation aiming for enhanced abiotic stress tolerance.