Identification of SPX family genes in the maize genome and their expression under different phosphate regimes

Many studies have revealed that SPX (SYG1/Pho81/XPR1) family genes play a key role in signal transduction related to phosphorus (P) deficiency in plants. Here, we identified 33 SPX gene family members in maize through genome-wide analysis and classified them into 4 subfamilies according to SPX structural characteristics (SPX, SPX-MFS, SPX-EXS and SPX-RING). The promoter regions of ZmSPXs are rich in biotic/abiotic-related stress elements. The quantitative real-time PCR analysis of 33 ZmSPXs revealed that all members except for ZmSPX3 of the SPX subfamily were significantly induced under P-deficient conditions, especially ZmSPX4.1 and ZmSPX4.2, which showed strong responses to low P stress and exhibited remarkably different expression patterns in low Pi sensitive and insensitive cultivars of maize. These results suggested that the SPX subfamily might play pivotal roles in P stress sensing and response. Experimental observations of subcellular localization in maize protoplasts indicated the following results, implying multiple roles in cell metabolism: ZmSPX2, ZmSPX5 and ZmSPX6 localized in the nucleus; ZmSPX1 and ZmSPX3 localized in the nucleus and cytoplasm; and ZmSPX4.2 localized in the chloroplast. A Y2H assay suggested that ZmPHR1 could interact with ZmSPX3, ZmSPX4.2, ZmSPX5, and ZmSPX6, indicating the involvement of these proteins in the P stress response in a ZmPHR1-mediated manner.