Interactions among rice ZMM proteins. A, Analysis of interactions between OsSHOC1 and ZMM proteins using the Y2H assay. B, Analysis of interactions between ZMM proteins using the Y2H assay. Full-length OsSHOC1, OsPTD, and ZMM protein coding sequences were constructed into both pGADT7 (Gal4AD) and PGBKT7 (Gal4BD) vectors. Selective-medium -LT (SD-Leu/-Trp), and -LTHA (SD-Leu/-Trp/-His/-Ade) with X-α-gal, were used to test interactions in the Y2H system. C, Diagram summarizing the network of interactions among ZMM proteins identified by Y2H system. Two-way arrows indicate interactions between rice ZMM proteins identified in this study (black) and those in previous reports (red). D and E, Y3H analysis among OsSHOC1, OsPTD, OsHEI10, and OsZIP4. Full-length OsSHOC1 and OsPTD protein coding sequences were constructed into pGADT7 vectors, and OsZIP4, OsHEI10, OsSHOC1, and OsPTD were separately cloned into multiple cloning sites I (MCSI) and II (MCSII) of pBridge vectors. Yeast cells were grown on -LTM (SD-Leu/-Trp/-Met) selection agar plates or -LTHAM (SD-Leu/-Trp/-His/-Ade/-Met) agar plates with X-α-gal for interaction detection.