Humic acid differentially improves nitrate kinetics under low- and high-affinity systems and alters the expression of plasma membrane H+-ATPases and nitrate transporters in rice

Abstract

Humic acids (HAs) have a major effect on nutrient uptake, metabolism, growth and development in plants. Here, we evaluated the effect of HA pretreatment applied with a nutrient solution on the uptake kinetics of nitrate nitrogen (N-NO3−) and the metabolism of nitrogen (N) in rice under conditions of high and low NO3− supply. In addition, the kinetic parameters of NO3− uptake, N metabolites, and nitrate transporters (NRTs) and the plasma membrane (PM) H+-ATPase gene expression were examined. The plants were grown in a growth chamber with modified Hoagland and Arnon solution until 21 days after germination (DAG), and they were then transferred to a solution without N for 48 h and then to another solution without N and with and without the addition of HAs for another 48 h. After this period of N deprivation, the plants received new nutrient solutions containing 0.2 and 2.0 mM N-NO3−. Treatment of rice plants with HA promoted the induction of the genes OsNRT2.1-2.2/OsNAR2.1 and some isoforms PM H+-ATPase in roots. The application of HAs differentially modified the parameters of the uptake kinetics of NO3− under both concentrations. When grown with 0.2 mM NO3−, the plants pretreated with HA had lower Km and Cmin values as well as a higher Vmax/Km ratio. When grown with 2 mM NO3−, the plants pretreated with HA had a higher Vmax value, a greater root and shoot mass, and a lower root/shoot ratio. The N fractions were also altered by pretreatment with HA, and a greater accumulation of NO3− and N-amino was observed in the roots and shoots, respectively, of plants pretreated with HA. The results suggest that pretreatment with HA modifies root morphology and gene expression of PM H+-ATPases and NO3− transporters, resulting in a greater efficiency of NO3− acquisition by high- and low-affinity systems.