The present invention relates to isolated male tissue-preferred regulatory sequences from Ms45 gene. The invention also relates to the use of Ms45 gene male tissue-preferred regulatory sequences in mediating male fertility in a plant, and in producing hybrid seed. The Ms45 male tissue-preferred regulatory region can be operably linked to exogenous genes encoding cytotoxins or inhibitory molecules. The invention also relates to plant cells, plant tissue and plants that contain the MS45 gene regulatory sequences.