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. 2000 Sep 26;97(21):11632–11637. doi: 10.1073/pnas.190309197

Figure 5.

Figure 5

Transactivation of the rd29B promoter-GUS fusion gene by AREB1 and AREB2 proteins using Arabidopsis protoplasts. (A) Expression of rd29B in response to dehydration, low temperature, high salt, or ABA in wild type (Columbia & Landsberg) or ABA-related mutants (aba2, abi1, and abi3). Plants were untreated (C), dehydrated for 10 h (D), or treated with water (H), 250 mM NaCl (N) or 100 μM ABA (A) for 5 h. (B) Schematic diagram of the effector and reporter constructs used in cotransfection experiments. The effector constructs contain the CaMV 35S promoter and TMV Ω sequence fused to AREB1, AREB2, or GBF3 cDNAs. The reporter constructs contain the 77-bp fragments of the rd29B promoter connected tandemly five times (×5) or single (×1). The promoters were fused to the −51 rd29B minimal TATA promoter-GUS construct. (C) Transactivation of the rd29B promoter-GUS fusion gene by AREB1, AREB2, and GBF3 proteins. The reporter gene driven by the 77-bp fragments tandemly repeated five times was transfected with each effector plasmid or the vector as control treatments. Transactivation experiments using protoplasts prepared from wild type or ABA-mutant (abi1, aba2, and era1) Arabidopsis leaves. To normalize for transfection efficiency, the CaMV 35S promoter-luciferase (LUC) plasmid was cotransfected in each experiment. Bars indicate the standard error of three replicates. Ratios indicate the multiples of expression compared with the value obtained with the pBI35SΩ vector, and ratio in parentheses indicates the multiples of expression compared with value obtained with the wild type transformed with pBI35SΩ vector. (D) Transactivation using the reporter construct containing the single 77-bp fragment.