OsHAD1, a haloacid dehalogenase-like APase enhances phosphate accumulation

Abstract

Phosphorus (P) deficiency limits plant growth and crop yield. Since, plants can absorb only inorganic form of P (Pi), a large portion of soil P (organic and inorganic P complexes) remains largely unused. Here, we identified and characterized a PHR2 regulated; novel low Pi responsive haloacid dehalogenase (HAD)-like hydrolase, OsHAD1. While, OsHAD1 is a functional HAD protein having both acid phosphatase and phytase activity; it showed little homology with other known low Pi responsive HAD superfamily members. Recombinant OsHAD1 is highly active at acidic pH and dephosphorylates broad range of organic and inorganic P containing substrates including protein phosphates and Na-phytate. Exogenous application of recombinant OsHAD1 protein in growth media supplemented with phytate, led to marked increase in growth and total P content of Pi deficient WT rice seedlings. Further, overexpression of OsHAD1 in rice resulted in enhanced phosphatase activity, biomass, total and soluble P content in Pi deficient transgenic seedlings treated with phytate as restricted Pi source. Gene expression and metabolite profiling revealed enhanced Pi starvation responses such as upregulation of multiple genes involved in Pi uptake and solubilization, accumulation of organic acids, enhanced secretory phosphatase activity and depletion of ATP in overexpression lines as compared to WT. To elucidate the underlying regulatory mechanisms of OsHAD1, we performed in-vitro pull down assays which revealed association of OsHAD1 with protein kinases. We conclude that besides dephosphorylation of cellular organic-P, OsHAD1 in coordination with kinases may regulate phosphorylation status of downstream targets to accomplish Pi homeostasis under limited Pi supply.