The mature pollen grain displays a highly specialized function in angiosperms. Accordingly, the male gametophyte development involves many specific biological activities, making it a complex and unique process in plants. In order to accomplish this, during pollen development, a massive transcriptomic remodeling takes place, indicating the switch from a sporophytic to a gametophytic program and involving the expression of many pollen specific genes. Using microarray databases we selected genes showing pollen-specific accumulation of their mRNAs and confirmed this through RT-PCR. We selected five genes (POLLEN SPECIFIC GENE1-5) to investigate the pollen specificity of their expression. Transcriptional fusions between the putative promoters of these genes and the uidA reporter gene in Arabidopsis confirmed the pollen specific expression for at least two of these genes. The expression of the cytotoxin Bamase controlled by these promoters generated pollen specific ablation and male sterility. Through the selection of pollen specific genes from public datasets, we were able to identify promoter regions that confer pollen expression. The use of the cytotoxin Barnase allowed us to demonstrate its expression is exclusively limited to the pollen. These new promoters provide a powerful tool for the expression of genes exclusively in pollen. (C) 2014 Elsevier Masson SAS. All rights reserved.