Self-incompatibility in solanaceous plants is gametophytically controlled by a multiallelic S-locus. The only known S-locus product is a series of extracellular ribonucleases (the S-RNases) which are expressed in the mature style and determine its self-incompatibility phenotype. Potentially, S-RNases also determine the self-incompatibility phenotype of pollen as some solanaceous plants express this protein for a brief period during anther development. To test this, we first showed that the S3-RNase of Lycopersicon peruvianum is expressed during anther development. We then transformed L. peruvianum plants with sense and antisense versions of the S3-RNase coding region under the control of a pollen-specific promoter. Pollen from the transgenic plants accumulated S3-RNase transcripts and the S3-RNase protein was detected immunologically in the sense transgenic plants. However, neither the sense nor the antisense S3-RNase constructs altered the self-incompatibility phenotype of pollen from the transgenic plants, demonstrating the S3-RNase is not the pollen product of the S3-allele.