An RNA-binding protein MUG13.4 interacts with AtAGO2 to modulate salinity tolerance in Arabidopsis

Salt stress is a major constraint to plant growth and development, and plants have developed sophisticated mechanisms to cope with it. AtAGO2, an argonaute protein, is known to play an important role in plant adaptation to salt stress; however, the molecular mechanism of this phenomenon remains essentially unexplored. Here, we performed the yeast two-hybrid assay and found an R3H-domain containing protein, designated as MUG13.4, interacting with AtAGO2. Further bimolecular fluorescence complement (BiFC), glutathione-S-transferase (GST) pull-down, and co-immunoprecipitation (Co-IP) assays confirmed that MUG13.4 interacted with AtAGO2, and MUG13.4 could affect the slicing activity of AtAGO2 associated with miR173. MUG13.4 and AtAGO2 were both predominantly expressed in seeds and roots. Phenotypic analyses of the single and double mutants under salt stress confirmed involvement of MUG13.4-AtAGO2 complex as a component of the salt tolerance mechanism. The mug13.4×ago2-1 double mutant displayed retarded growth and hypersensitivity to salt stress that was more pronounced than in mug13.4 or atago2-1 single mutants. TAS1c-tasiRNA generating system in Nicotiana benthamiana revealed that MUG13.4 could influence the slicing activity of AtAGO2. We also found that MUG13.4 increasingly changed the phenotype of slicer-defected mutants of AtAGO2 in response to salt stress. These findings suggested that the function of AtAGO2 upon salt stress was dependent on MUG13.4. Further investigation suggested that AtAGO2 improved Arabidopsis tolerance to salt stress by affecting operation of the SOS signaling cascade at the transcript level. Taken together, these findings reveal a novel function of MUG13.4 in adjusting Arabidopsis adaptation to salt stress.