Agrobacterium tumefaciens-mediated genetic transformation is a powerful tool for plant research,but it
can be labor-intensive and time-consuming.Here, we report a protoplast-based approach to study
nucleo-cytoplasmic interactions,such as cytoplasmic male sterility/fertility restoration (CMS/Rf) and
organellar RNA editing. To test the system,we transfected the fertility restorer gene Rf5, which is involved
in the rice HL-CMS/Rf system, into rice protoplasts prepared from the HL-CMS line. As the Rf5 protein
accumulated in the transformed protoplasts,the CMS-associated transcripts were endonucleolytically
cleaved.There were much lower levels of the CMS-associated protein ORFH79 in the transfected protoplasts than in the mock-transfected protoplasts.