Novel underlying regulatory mechanism of the MsDAD2-mediated salt stress response in alfalfa

Alfalfa (Medicago sativa L.), a crucial and widely grown forage legume, faces yield and quality challenges due to salinity stress. The defender against apoptotic death (DAD) gene, recognized initially as an apoptosis suppressor in mammals, plays a pivotal role in catalyzing N-glycosylation, acting as a positive regulator for protein folding and endoplasmic reticulum (ER) export. Here, we found that the MsDAD2 gene was specially induced in the salt tolerant alfalfa cultivar (DL) under salinity stress, but not in the salt-sensitive cultivar (SD). Overexpression of MsDAD2 enhanced the salinity resistance of transgenic alfalfa by promoting NAD(P)H-quinone oxidoreductase (NQO1) and cytochrome b6f complex subunit (Cyt b6/f) expression, thereby mitigating reactive oxygen species (ROS) production. ChIP-qPCR analysis suggested that the differential expression of MsDAD2 in DL and SD under salinity stress may be linked to dynamic histone modifications in its promoter. Therefore, our findings elucidate a novel regulatory mechanism of MsDAD2 in alfalfa's response to salinity stress, underscoring its significance as a target for alfalfa breeding to enhance salt tolerance.

Automated summary

The MsDAD2 gene was specifically induced in the salt tolerant alfalfa cultivar and its overexpression enhanced the salinity resistance of transgenic alfalfa by promoting the expression of NQO1 and Cyt b6/f, leading to reduced reactive oxygen species production. The expression difference of MsDAD2 may be linked to dynamic histone modifications in its promoter. This study highlights the importance of MsDAD2 as a target for improving salt tolerance in alfalfa breeding.