OsCaM1‐1 Is Responsible for Salt Tolerance by Regulating Na⁺/K⁺ Homoeostasis in Rice

Calmodulin, a highly conserved calcium-binding protein, plays a crucial role in response to salt stress. Previous studies investigated sequence and function of calmodulin members in some plants, but their roles in rice have not been fully elucidated. Three OsCaM1 genes namely OsCaM1-1/2/3 encode the same OsCaM1 protein. Here, we found that OsCaM1-1 had significantly higher expression than the other two genes under salt stress. After 4 weeks of exposure to 75 mM NaCl, OsCaM1-1 overexpressed mutants showed higher salt tolerance, while knocked-out mutants exhibited lower salt tolerance, compared to the wild type. Moreover, the oscam1-1 mutants had higher Na⁺ concentration and Na⁺/K⁺ ratio in both shoots and roots, less instantaneous K⁺ and Ca²⁺ fluxes in roots, compared to wild type under salt stress, indicating the involvement of OsCaM1-1 in regulation of Na⁺ and K⁺ homoeostasis via Ca²⁺ signal. RNA-seq analysis identified 452 differentially expressed genes (DEGs) regulated by OsCaM1-1 and salt stress, and they were mainly enriched in nucleus DNA-binding activities, including ABI5, WRKY76, WRKY48 and bHLH120 transcription factors. Knockout of OsCaM1-1 also modulated the expression of Na⁺ transporters, including HKT1;1, HKT1;5, SOS1, NHX1 and NHX4. In conclusion, OsCaM1-1 positively regulates salt tolerance in rice through mediating ion homoeostasis.