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. 2016 Mar 29;6:23609. doi: 10.1038/srep23609

Figure 2. ANAC046 activates the promoters of chlorophyll catabolic genes.

Figure 2

(a) Activation of NAC transcription factors against promoters of chlorophyll catabolic genes was tested in Y1H experiment. The following promoters were tested: NYC1, SGR1, SGR2, and PaO. The transcription factors tested were as follows: 1, none (empty vector pGAD424); 2, ANAC046; 3, ANAC087; 4, ORE1; 5, ORS1; 6, ANAC079; and 7, ANAC100. Growth of yeast transformants carrying the HIS3 reporter gene under the control of each promoter region was examined in the medium (SD) lacking histidine (H) and leucine (L) but containing 3-AT. (b) Transactivation of the Chl catabolic gene promoter–firefly luciferase fusion genes by the ANAC046 protein in Arabidopsis leaf protoplasts. The LUC reporter gene under the control of the Chl catabolic gene promoters was co-transfected with an effector plasmid expressing ANAC046-VP16 or VAMP722-VP16 (negative control). To normalize transfection efficiency, the CaMV 35S promoter–Renilla luciferase plasmid was cotransfected in each experiment and firefly luciferase activity was normalized to that of Renilla luciferase. Gene designations are as in Fig. 1. Bars indicate standard error (4 biological replicates). Asterisks indicate significant differences (P < 0.01) between VAMP722-VP and ANAC046-VP according to Student’s t-test.