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Small peptides secreted to the extracellular matrix control many aspects of the plant’s physiological activities which were identified in  Arabidopsis thaliana , called ATSPs. Here, we isolated and characterized the small peptide gene  Bna.SP6  from  Brassica napus . The  BnaC.SP6  promoter was cloned and identified. Promoter deletion analysis suggested that the -447 to -375 and -210 to -135 regions are crucial for the silique septum and pollen expression of  BnaC.SP6 , respectively. Furthermore, the minimal promoter region of p158 (-210 to -52) was sufficient for driving gene expression specifically in pollen and highly conserved in  Brassica  species. In addition,  BnaA.bZIP1  was predominantly expressed in anthers where  BnaC.SP6  was also expressed, and was localized to the nuclei. BnaA.bZIP1 possessed transcriptional activation activity in yeast and protoplast system. It could specifically bind to the C-box in p158  in vitro , and negatively regulate p158 activity  in vivo . BnaA.bZIP1 functions as a transcriptional repressor of  BnaC.SP6  in pollen activity. These results provide novel insight into the transcriptional regulation of  BnaC.SP6  in pollen activity and the pollen/anther-specific promoter regions of  BnaC.SP6  may have their potential agricultural application for new male sterility line generation.

BnaA.bZIP1 Negatively Regulates a Novel Small Peptide Gene, BnaC.SP6, Involved in Pollen Activity