Gene duplication has been recognized as a major route to supply raw sequences for genome novelty in evolution, but the mechanism underlying the retention of duplicate genes is not fully understood yet. Divergence in spatial expression patterns was investigated here and in response to stimuli of the four members of a rice proline-rich protein gene family (OsPRP1), which encode a class of proline-rich proteins. The four paralogues are tandemly organized within a 20 kbp range of chromosome 10 without any interval of other open reading frames and with a median K(S) value of 0.474. These paralogues showed little similarity in their regulatory regions but high conservation in coding regions. Search of an intergenomic cis-element database predicted their promoter regions with divergent cis-element fingerprints. Further expression analyses involving different tissues/organs and nine types of stimuli by a promoter::GUS-fusion strategy revealed that the four paralogues were expressed mainly in vascular cylinders of different organs and showed diversity in tissue/organ specificity and in response to these stimuli, with some overlapping expression. Furthermore, these data show that OsPRP1.2 appeared to inherit most of the functions from their multifunctional progenitor, whereas the other three genes diverged after duplication events. Thus, the retention of paralogues in a multigene family seems to require a more complicated diversification process than originally thought. In addition, the promoter::GUS strategy is a powerful way to explore function divergence of a tandem-repeat gene family.