Nitrate transporter NRT1.1 and anion channel SLAH3 form a functional unit to regulate nitrate-dependent alleviation...

Ammonium (NH₄ ⁺ ) and nitrate (NO₃ ^- ) are major inorganic nitrogen (N) sources for plants. When serving as the sole or dominant N supply, NH₄ ⁺ often causes root inhibition and shoot chlorosis in plants, known as ammonium toxicity. NO₃ ^- usually causes no toxicity and can mitigate ammonium toxicity even at low concentrations, referred to as nitrate-dependent alleviation of ammonium toxicity. Our previous studies indicated a NO₃ ^- efflux channel SLAH3 is involved in this process. However, whether additional components contribute to NO₃ ^- -mediated NH₄ ⁺ detoxification is unknown. Previously, mutations in NO₃ ^- transporter NRT1.1 were shown to cause enhanced resistance to high concentrations of NH₄ ⁺ . Whereas, in this study, we found when the high-NH₄ ⁺ medium was supplemented with low concentrations of NO₃ ^- , nrt1.1 mutant plants showed hyper-sensitive phenotype instead. Furthermore, mutation in NRT1.1 caused enhanced medium acidification under high-NH₄ ⁺ /low-NO₃ ^- condition, suggesting NRT1.1 regulates ammonium toxicity by facilitating H⁺ uptake. Moreover, NRT1.1 was shown to interact with SLAH3 to form a transporter-channel complex. Interestingly, SLAH3 appeared to affect NO₃ ^- influx while NRT1.1 influenced NO₃ ^- efflux, suggesting NRT1.1 and SLAH3 regulate each other at protein and/or gene expression levels. Our study thus revealed NRT1.1 and SLAH3 form a functional unit to regulate nitrate-dependent alleviation of ammonium toxicity through regulating NO₃ ^- transport and balancing rhizosphere acidification.