Overexpression of a microRNA-targeted NAC transcription factor improves drought and salt tolerance in Rice via ABA-mediated pathways

Background: The NAC (NAM, AFAT, and CUC) transcription factors play critical roles in rice (Oryza sativa) development and stress regulation. Overexpressing a microRNA (miR164b)-resistant OsNAC2 mutant gene, which generates transcripts that cannot be targeted by miR164b, improves rice plant architecture and yield; however, the performance of these mOsNAC2-overexpressing lines, named ZUOErN3 and ZUOErN4, under abiotic stress conditions such as drought have not yet been fully characterized.

Results: In this study, we showed that the germination of ZUOErN3 and ZUOErN4 seeds was delayed in comparison with the wild-type (WT) seeds, although the final germination rates of all lines were over 95%. The quantification of the endogenous ABA levels revealed that the germinating mOsNAC2-overexpressing seeds had elevated ABA levels, which resulted in their slower germination. The mOsNAC2-overexpressing plants were significantly more drought tolerance than the WT plants, with the survival rate increasing from 11.2% in the WT to nearly 70% in ZUOErN3 and ZUOErN4 plants after a drought treatment. Salt (NaCl) tolerance was also increased in the ZUOErN3 and ZUOErN4 plants due to significantly increased ABA levels. A reverse transcription quantitative PCR (RT-qPCR) analysis showed a significant increase in the expression of the ABA biosynthesis genes OsNCED1 and OsNCED3 in the mOsNAC2-overexpressing lines, and the expression levels of the stress-responsive genes OsP5CS1, OsLEA3, and OsRab16 were significantly increased in these plants. Moreover, OsNAC2 directly interacted with the promoters of OsLEA3 and OsNCED3 in yeast one-hybrid assays.

Conclusions: Taken together, our results show that OsNAC2 plays a positive regulatory role in drought and salt tolerance in rice through ABA-mediated pathways.