Oryza sativa, C4HC3-type really interesting new gene (RING), OsRFPv6, is a positive regulator in response to salt stress by regulating Na+ absorption

Abstract

Salinity negatively affects plant growth, productivity, and metabolism. Therefore, plants have evolved diverse strategies to survive in saline environments. To identify such strategies involving the ubiquitin/26S proteasome system, we characterized molecular functions of a rice C4HC3 really interesting new gene (RING)-type E3-ubiquitin ligase gene. Oryza sativa RING finger protein v6 (OsRFPv6) was highly expressed under conditions of abiotic stress, induced by 100 mM NaCl and 20% PEG. The GFP-OsRFPv6 protein was localized in the plasma membrane and cytosol in rice protoplasts. In vitro ubiquitin assay revealed that OsRFPv6 possessed E3-ubiquitin ligase activity, but its variant OsRFPv6C100A did not. OsRFPv6-overexpressing plants were insensitive to salinity, but their growth was delayed under normal conditions. Under saline conditions, transgenic plants exhibited higher proline, soluble sugar, and chlorophyll content and lower H₂ O₂ accumulation than wild-type plants. Moreover, transgenic plants exhibited lower Na⁺ uptake, lower Na⁺ content, and higher K⁺ content in the xylem sap assay. Under saline conditions, the expression levels of nine Na⁺ /K⁺ transporter genes in roots and leaves were significantly different between transgenic and wild-type plants. Specifically, under both normal and saline conditions, the expression of OsHKT2;1, a Na⁺ transporter, in the roots of transgenic plants was lower than that in the roots of wild-type plants. These results suggest that OsRFPv6 E3-ubiquitin ligase serves as a positive regulator of salinity response via Na⁺ uptake.