Processing of pre-rRNA is one of the major events taking place In the nucleolus. U3 snoRNA, an rRNA spliceosomal factor, is suggested to be essential in the first cleavage step of the 5' ETS sequence in the processing of pre-rRNA. Identification of U3 in the nucleolus provides a piece of indirect evidence for pre-rRNA processing site and transportation of processing products. In the present study, subnucleolar distribution of U3 snoRNA in the nucleolus of Pisum sativum L. was studied by in situ hybridization with a U3 snoRNA probe. The results showed that the U3 labeling signals were distributed throughout dense fibrillar components (DFCs) and granular components (GCs), while no signal was found in fibrillar centers (FCs). When treated with actinomycine D (AMD), the labeling signals were decreased. Along with the increase of the AMD treatment time, the labeling signals became fewer and they were found in the distal regions of DFC and GC. Our results indicated that pre-rRNA splicing took place in the regions of DFC and GC, and the transportation of pre-rRNA processing products was from the regions around FCs towards the distal regions.