Knockdown of OsPHP1 Leads to Improved Yield Under Salinity and Drought in Rice via Regulating the Complex Set of TCS Members and Cytokinin Signalling

Plant two-component system (TCS) is crucial for phytohormone signalling, stress response, and circadian rhythms, yet the precise role of most of the family members in rice remain poorly understood. In this study, we investigated the function of OsPHP1, a pseudo-histidine phosphotransfer protein in rice, using a functional genomics approach. OsPHP1 is localised in the nucleus and cytosol, and it exhibits strong interactions with all sensory histidine kinase proteins (OsHK1-6) and cytokinin catabolism genes. Our results demonstrate that OsPHP1 functions as a negative regulator of cytokinin signalling. Knockdown of OsPHP1 enhanced the expression of positive cytokinin signalling regulators, such as OsHKs and OsAHPs (authentic phosphotransfer proteins), while downregulating negative regulators, such as type-A response regulators (OsRRs) and cytokinin catabolism genes (CKXs). Furthermore, OsPHP1 negatively influences abiotic stress tolerance, as evidenced by the increased sensitivity of OsPHP1-OE (overexpression) lines to salinity and drought. In contrast, OsPHP1-KD (knockdown) lines showed enhanced stress resilience, with better photosynthesis, increased tiller and panicle production, higher spikelet fertility, and grain filling. The study demonstrates that OsPHP1 suppresses antioxidant and stress-responsive genes, exacerbating ion toxicity and reducing osmolyte accumulation, thereby impairing plant growth and yield under stress conditions. These findings highlight OsPHP1 as a critical modulator of plant responses to abiotic stress and suggest potential genetic targets for enhancing crop stress tolerance.