The maize (Zea mays) late pollen gene ZmMADS2 belongs to the MIKC type of MADS box transcription factor genes. Here, we report that ZmMADS2, which forms a homodimer in yeast (Saccharomyces cerevisiae), is required for anther dehiscence and pollen maturation. Development of anthers and pollen was arrested at 1 d before dehiscence in transgenic plants expressing the ZmMADS2-cDNA in antisense orientation. Temporal and spatial expression analyses showed high amounts of ZmMADS2 transcripts in endothecium and connective tissues of the anther at 1 d before dehiscence and in mature pollen after dehiscence. Transient transformation of maize and tobacco (Nicotiana tabacum) pollen with the luciferase reporter gene under the control of different ZmMADS2 promoter deletion constructs demonstrated the functionality and tissue specificity of the promoter. Transgenic maize plants expressing a ZmMADS2-green fluorescent protein fusion protein under control of the ZmMADS2 promoter were used to monitor protein localization during anther maturation and pollen tube growth. High amounts of the fusion protein accumulate in degenerating nuclei of endothecial and connective cells of the anther. A possible function of ZmMADS2 during anther dehiscence and pollen maturation and during pollen tube growth is discussed.