We fused the N-terminus of TurboID to the chloroplast outer membrane protein OEP7 and the C-terminus to the endoplasmic reticulum membrane protein RTNLB2. When the chloroplast and endoplasmic reticulum come into close proximity to form membrane contact sites (MCSs), the split-TurboID is reconstituted into an active proximity-labeling enzyme that biotinylates nearby proteins. In this proteomic study, Barley stripe mosaic virus (BSMV) infection was used as the experimental variable.