Regulation of tapetum-specific A9 promoter by transcription factors AtMYB80, AtMYB1 and AtMYB4 in Arabidopsis thaliana and Nicotiana tabacum.

Tapetum-specific promoters have been successfully used for developing transgenic-based pollination control systems. Although several tapetum-specific promoters have been identified, in-depth studies on regulation of such promoters are scarce. The present study analyzes the regulation of the A9 promoter, one of the first tapetum-specific promoter identified in Arabidopsis thaliana. Transcription factors (TFs) AtMYB80, AtMYB1 (positive regulators) identified by in silico analysis were found to upregulate A9 promoter activity following the over-expression of the TFs in transient and stable (transgenic) expression assays in both A. thaliana and tobacco. Furthermore, mutations of binding sites of these TFs in the A9 promoter led to loss of its activity. The role of a negative regulator AtMYB4 was also studied by analyzing the activity of A9 promoter following transient expression of RNAi against the TF and by mutating binding sites for AtMYB4 in the A9 promoter. While no changes were observed in case of A. thaliana, the A9 promoter was activated in the roots of transgenic tobacco plants, highlighting the role of these cis-elements in keeping the A9 promoter repressed in the roots of tobacco.