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. 2011 Mar 30;156(2):537–549. doi: 10.1104/pp.111.177071

Figure 6.

Figure 6.

The IAA30 gene is involved in the NTM2-mediated salt regulation of germination. In A to D, transcript levels were determined by qRT-PCR. Biological triplicates were averaged. Error bars indicate se. Statistical significance was determined by Student’s t test (* P < 0.01). Two-week-old plants grown on MS-agar plates were used for treatments with NaCl and IAA. Whole plants were used for extraction of total RNA. M, Mock; S, NaCl. A, Effects of high salinity on IAA gene transcription. Plants were transferred to MS liquid cultures supplemented with 150 mm NaCl and soaked for 3 h before harvesting plant materials. B, Kinetic expression pattern of the IAA30 gene under high salinity. Plants were soaked in MS liquid cultures supplemented with 150 mm NaCl for the indicated time periods before harvesting plant materials. C, IAA30 gene transcription in germinating seeds under high salinity. Cold-imbibed seeds were allowed to germinate for 30 h, and total RNA was extracted from the germinating seeds. D, Kinetic expression pattern of the IAA30 gene after auxin treatments. Plants were transferred to MS liquid cultures supplemented with 10 μm IAA and incubated for the indicated time periods before harvesting plant materials. E, Germination phenotypes of the ntm2-1 mutant overexpressing the IAA30 gene. The ntm2-1 mutant was transformed with the IAA30 gene driven by the CaMV 35S promoter (30OX/ntm2-1). Germination assays were carried out as described in Figure 4B. F, Electrophoretic mobility shift assays on ΔC binding to a conserved sequence in the IAA30 gene promoter. The recombinant ΔC protein was prepared as a MBP-ΔC fusion in E. coli cells. The minus (–) lane is a control without adding ΔC protein. Approximately 0.5 μg of the ΔC protein was used for each assay. The conserved nucleotides underlined were mutated in the mIAA30-BS sequence. Unlabeled DNA fragments were added as competitors. G, Transcriptional activation activity assays in Arabidopsis protoplasts. The IAA30-BS or mIAA30-BS sequence was fused upstream to a 35S minimal TATA promoter (Min35S)-GUS reporter construct (top panel). The reporter and the 35S:ΔC effector constructs were cotransformed into Arabidopsis protoplasts. Luciferase gene expression was used to normalize the GUS activity. Five measurements were averaged (bottom panel). Error bars indicate se. Statistical significance was determined by Student’s t test (* P < 0.01). Nos, Terminator in the pCAMBIA 1305.1 vector.