Mecanismos cinéticos de inibição da absorção de nitrato em raízes de feijão caupi expostas à salinidade.

Abstract

Nitrate uptake is a process dependent on the influx/ efflux relationship in the plasma membrane of roots cells. The balance between these two processes may affect the NO3- acquisition in plants exposed to salt stress. Unfortunately, we do not know the mechanisms of inhibition of NO3- uptake in the presence of excess Na + and Cl- ions, a common situation in saline-sodium soil. The objective was to evaluate the effect of salinity on the NO3- influx and efflux mechanisms in cowpea roots, building the uptake kinetics in the presence of ion and osmotic agents. Cowpea roots were excised for nitrate influx and efflux measurements in time-course and NO3- levels experiments. In plants pre-treated with 50 and 100 mM NaCl, NO3- influx and eflux kinetics has been altered, compared with plants not pre-treated roots: inducing pronounced effects on both processes, inhibiting the influx and stimulating the efflux. The NaCl inhibitory effect on the NO3- influx was dependent on the dose of salt. The 100 mM NaCl reduced the value of Vmax (control) of 12.35 to 7.14 and the Km value remained unchanged (3.05 and 3.21, respectively), showing a non-competitive inhibition. Interestingly, the Na2SO4 100 mM caused the same inhibition of NaCl, but 100 mM KCl caused an incompetitive inhibition, reducing Vmax and Km. In order to check the osmotic effects components by salinity, it was observed that PEG and Mannitol caused a competitive inhibition, increasing the Km (from 3.01(control) to 6.21 7.35 for PEG and Mannitol, respectively) and maintaining Vmax below the control. The NaCl inhibitory effect was more intense on NO3- uptake low affinity system (60%) than on the high-affinity system (45%). The NaCl and mannitol effects on the high affinity system, both in 100 mM, were similar to each other inhibition type: Vmax reduced both and increased the Km. When NaCl, PEG and mannitol isosmotic concentrations were tested (Ψos= - 0.23 MPa), it was found that the first two induced efflux increases (15 and 34% respectively), whereas, unexpectedly, Mannitol caused a reduction of 40%, all compared to control. In conclusion, the salinity caused reduction in net NO3- uptake in cowpea roots through reduction in the influx and efflux increases. In the influx, there is a non competitive inhibition with the Cl-in both low and high affinity systems. Apparently, the osmotic component is more important inhibiting the NO3- uptake, but apparently, the salt type (ions) also interferes in the process. Meanwhile, more studies are needed to shed light on the mechanisms involved in inhibition.