Sugarcane streak mosaic virus (SCSMV) is one of the causative agents of sugarcane (Saccharum officinarum L.) mosaic disease.In this study a sugarcane cDNA library was screened with P3 from SCSMV as bait using yeast two-hybrid system (Y2HS).A gene encoding the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) was obtained and designed as ScRbcL.The open reading frame (ORF) of ScRbcL is 1 337 bp in length and codes for 478 amino acids.The interaction between the ScRbcL and SCSMV-P3 was identified by Y2HS and bimolecular fluorescence complementation (BiFC).This research provided new cues to the investigation of the molecular mechanism of SCSMV infection on sugarcane,especially on the mosaic syndrome development caused by sugarcane mosaic disease.