Endogenous ABA alleviates rice ammonium toxicity by reducing ROS and free ammonium via regulation of the SAPK9–bZIP20 pathway

Ammonium (NH(4)(+)) is one of the principal nitrogen (N) sources in soils, but is typically toxic already at intermediate concentrations. The phytohormone abscisic acid (ABA) plays a pivotal role in responses to environmental stresses. However, the role of ABA under high-NH(4)(+) stress in rice (Oryza sativa L.) is only marginally understood. Here, we report that elevated NH(4)(+) can significantly accelerate tissue ABA accumulation. Mutants with high (Osaba8ox) and low levels of ABA (Osphs3-1) exhibit elevated tolerance or sensitivity to high-NH(4)(+) stress, respectively. Furthermore, ABA can decrease NH(4)(+)-induced oxidative damage and tissue NH(4)(+) accumulation by enhancing antioxidant and glutamine synthetase (GS)/glutamate synthetasae (GOGAT) enzyme activities. Using RNA sequencing and quantitative real-time PCR approaches, we ascertain that two genes, OsSAPK9 and OsbZIP20, are induced both by high NH(4)(+) and by ABA. Our data indicate that OsSAPK9 interacts with OsbZIP20, and can phosphorylate OsbZIP20 and activate its function. When OsSAPK9 or OsbZIP20 are knocked out in rice, ABA-mediated antioxidant and GS/GOGAT activity enhancement under high-NH(4)(+) stress disappear, and the two mutants are more sensitive to high-NH(4)(+) stress compared with their wild types. Taken together, our results suggest that ABA plays a positive role in regulating the OsSAPK9–OsbZIP20 pathway in rice to increase tolerance to high-NH(4)(+) stress.