We screened a cDNA library of mature maize pollen for clones encoding transcription factors. Two novel cDNAs (ZmMADS1 and ZmMADS2) encoding MADS box proteins were isolated. To our knowledge, ZmMADS1 and ZmMADS2 represent the first transcription factors isolated from pollen of a monocot plant species. ZmMADS1 can be classified as a member of the TM3 subfamily of MADS box proteins whereas ZmMADS2, which is described here in more detail, belongs to the AGL17 subfamily. Temporal and spatial expression analyses showed that ZmMADS2 expression is restricted to pollen and roots. During pollen development ZmMADS2 is detectable at all stages of pollen development analyzed by RT-PCR but is most abundant in mature pollen after dehiscence. The establishment of a whole-mount RNA in situ hybridization protocol with in-vitro germinated pollen enabled the localization of ZmMADS2 transcripts in pollen tubes. ZmMADS2 transcripts are translocated into pollen tubes immediately upon germination and display a gradient with increasing concentration towards the tube tip. The vegetative nucleus and the sperm cells, putative targets of ZmMADS2, enter the pollen tube about 2 h later.