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(a–c) Relative mRNA levels of OsNAC6 (a), AK104277 (b) and AK110725 (c) are shown.

Fourteen-day-old plants were used to isolate RNA. (i) Relative mRNA level of the genes in the untreated OsNAC6-OX rice plants compared with the mRNA level in the untreated vector control plants. (ii) Relative mRNA level of the genes in the 24 h DEX-treated OsNAC6-GR rice plants compared with the mRNA level in the 24 h DEX-treated vector control plants. (iii) Relative mRNA level of the genes in the POsNAC6-OsNAC6 and the PLIP9-OsNAC6 rice plants treated with 250 mM NaCl for 24 h compared with the mRNA levels in control plants exposed to the same saltstress treatment. (iv) Relative mRNA level of the genes in the stress-treated plants compared with the mRNA level in the untreated plants. Rice seedlings (Nipponbare) were grown hydroponically for 14 days and were then subjected to dry, high-salt (NaCl) and cold stresses for 24 h, and used to prepare total RNAs. (d) Activation of the promoter–GUS fusion gene by OsNAC6 using rice protoplasts. Rice protoplasts were transfected by the reporter plasmids using various sets of effector plasmids [vector containing ubiquitin promoter as a control (ubi-vector) and ubiquitin promoter–OsNAC6 (ubi-OsNAC6)] and reporter plasmids [the AK104277 promoter–GUS fusion (AK104277) and the AK110725 promoter–GUS fusion (AK110725)]. Co-transfection of a constitutively expressed luciferase (LUC) gene using the ubiquitin promoter allowed normalization of expression in independent experiments. Bars indicate the fold of the GUS activity compared with the reporter activity using the ubi-vector.