Functional analysis of the buckwheat metallothionein promoter: Tissue specificity pattern and up-regulation under complex stress stimuli

To shed light on expression regulation of the metallothionein gene from buckwheat (FeMT3), functional promoter analysis was performed with a complete 5′ regulatory region and two deletion variants, employing stably transformed tobacco plants. Histochemical GUS assay of transgenic tobacco lines showed the strongest signals in vascular elements of leaves and in pollen grains, while somewhat weaker staining was observed in the roots of mature plants. This tissue specificity pattern implies a possible function of buckwheat MT3 in those tissues. Quantitative GUS assay showed strong up-regulation of all three promoter constructs (proportional to the length of the regulatory region) in leaves submerged in liquid MS medium containing sucrose, after a prolonged time period. This represented a complex stress situation composed of several synergistically related stress stimuli. These findings suggest complex transcriptional regulation of FeMT3, requiring interactions among a number of different factors.