Fig. 6

PbC2H2.K16.2 is the upstream factor of PbCOB.A.1 and PbCOB.A.2. a Reporters and effector. b The LUC activities driven by the PbCOB.A.1 or PbCOB.A.2 promoter in tobacco leaves over-expressing PbC2H2.K16.2 were evaluated using a dual-luciferase assay. OE-replicate 1 and 2 indicate the two independent experiments. Standard error was calculated from at least five replicates. Analysis of variance was calculated by Student’s t-test. Asterisk indicate P < 0.05. EMSA assay showing the physical binding of PbC2H2.K16.2 to the PbCOB.A.1 (c) and PbCOB.A.2 promoters (d). ‘–’ and ‘ + ’indicate the absence and presence of the recombinant PbC2H2.K16.2-His protein, biotin-labeled probe, biotin-labeled mutant, or cold probe, respectively. Cold probe concentrations were tenfold ( +) and 100-fold (+ +) of labeled probes. MST assay showing the binding of PbC2H2.K16.2 to the PbCOB.A.1 (e) and PbCOB.A.2 promoters (f). X-axis represent the concentration gradients of DNA probe, while Y-axis represent the binding capability. The green, cyan, and red color dots represent the three replicates. Each dot represents the binding capacity of PbC2H2.K16.2 to the probe