Transcription factor OsNAC016 negatively regulates phosphate-starvation response in rice

Phosphate (Pi), the main form of inorganic phosphorus that can be absorbed by plants, is one of the most limiting macro-nutrients in plants. However, the underlying molecular mechanism determining how plants sense external Pi levels and reprogram transcriptional and adaptive responses is incompletely understood. At present, few rice NAC members have been reported to be involved in the signaling pathways of Pi homeostasis in plants. Here, our research demonstrated that OsNAC016, a Pi-starvation responsive gene in rice, was regulated by PHOSPHATE STARVATION RESPONSE protein 1 (OsPHR1) and OsPHR4. Under Pi-starvation stress, the root growth of OsNAC016-overexpression lines was inhibited more severely, and overexpression plants had lower Pi content than wild type, while osnac016 mutant was hyposensitive to Pi starvation, indicating that OsNAC016 negatively modulates rice Pi-starvation response. Chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR) analysis and transient transactivation assays indicated that OsNAC016 could activate the SPX-domain-containing protein 2 (OsSPX2) gene through binding to its promoter. Further, we found that Pi starvation enhanced OsNAC016 binding to the OsSPX2 promoter, thus strongly promoting OsSPX2 expression. At the same time, Pi starvation induced OsNAC016 protein accumulation in plants. Moreover, similar to OsSPX2, OsNAC016 negatively regulates leaf inclination by repressing the cell elongation in lamina joint in rice under Pi-starvation stress. Together, our findings demonstrate that OsNAC016 negatively regulates rice phosphate-starvation response and leaf inclination by activating OsSPX2 expression under Pi-starvation conditions. These data provide a strategy to create smart crops with ideal shoot architecture and high phosphorus utilization efficiency.