Calcium/calmodulin-dependent protein kinase OsDMI3 positively regulates saline-alkaline tolerance in rice roots

Soil saline-alkalization is a major environmental stress that impairs plant growth and crop productivity. Plant roots are the primary site for the perception of soil stresses; however, the regulation mechanism engaged in the saline-alkaline stress response in plant roots is not well understood. In this study, we identified how a rice Ca²⁺/calmodulin-dependent protein kinase, OsDMI3, confers saline-alkaline tolerance in rice root growth. We measured the OsDMI3 activity by an in-gel kinase assay, Na⁺ content by NaHCO₃ treatment, and Na⁺ and H⁺ fluxes by noninvasive micro-test technology (NMT). Furthermore, a real-time reverse-transcription polymerase chain reaction (RT-PCR) analysis was performed to identify the genes upregulated in response to NaHCO₃ treatment in rice roots. The results showed that NaHCO₃ significantly increased OsDMI3 expression and activity in rice roots. This was consistent with the results of Na⁺ content and NMT that indicated OsDMI3 promoted root elongation under saline-alkaline stress by reducing root Na⁺ and H⁺ influx. Moreover, real-time RT-PCR analysis revealed that OsDMI3 up-regulated the transcript levels of OsSOS1 and PM-H⁺-ATPase genes OsA3 and OsA8 in saline-alkaline stressed rice plants. Collectively, our results suggest that OsDMI3 could promote saline-alkaline tolerance in rice roots by modulating the Na⁺ and H⁺ influx. These findings provide an important genetic target for protection of growth in rice exposed to saline-alkaline stress.