Nitrate-dependent salt tolerance mediated by OsNLP4-OsMADS27 module

Salt stress is a major constraint of plant growth and yield. Nitrogen (N) fertilizers are known to alleviate salt stress. However, the underlying molecular mechanisms remain unclear. Here we show that OsNLP4-OsMADS27 module controls nitrate-dependent salt tolerance in rice. The expression of OsMADS27 is specifically induced by nitrate. The OsMADS27 knockout mutants are more sensitive to salt stress than the wild type, whereas the OsMADS27 overexpression lines are more tolerant. Transcriptomic analyses revealed that OsMADS27 controls the expression of a number of known stress-responsive genes as well as those involved in ion homeostasis and antioxidation. We demonstrated that OsMADS27 directly binds to the promoter of OsHKT1.1 and OsSPL7 to regulate their expression. Notably, OsMADS27-mediated salt tolerance is nitrate-dependent and positively correlated with nitrate concentration. We further showed that OsNLP4, a nitrate-responsive key regulator in N metabolism and N use efficiency, positively regulates the expression of OsMADS27 by directly binding to the nitrate-responsive cis-element in its promoter, thereby transmitting the nitrate signal to OsMADS27 and conferring its nitrate dependence. Our results reveal the role of nitrate-responsive OsNLP4-OsMADS27 module and its downstream target genes in salt tolerance, filling the gap in the molecular mechanism of nitrate-dependent salt tolerance of rice. Moreover, OsMADS27 overexpression increased grain yield under salt stress in presence of sufficient nitrate, indicating that OsMADS27 is a promising candidate for the improvement of salt tolerance in rice.