Net proton secretion as a parameter for nitrate uptake

Summary

Nitrate uptake and its link to net proton secretion in wheat (Triticum aestivum L. cv. Tassilo, Caribo, and Astron) were investigated using a pH-stat system. Since nitrate is taken up in symport with protons, nitrate and proton fluxes should be correlated. Nitrate concentration in the medium, measured by HPLC, decreased in a linear manner. The addition of nitrate caused a drop in net proton secretion rate to negative values (net proton influx). Once nitrate concentration had been lowered to a well defined level, net proton secretion rate started to recover. This critical nitrate concentration depended on the initial nitrate concentration in the medium. A technique to derive nitrate uptake rates from time courses of net proton secretion was developed and is described. Briefly, this method requires the initial nitrate concentration and the time until the minimal net proton secretion rate is achieved. Results determined with this technique were found in excellent agreement to simultaneous direct measurements of nitrate uptake by HPLC. Measurement of net proton secretion therefore can be used as a parameter for nitrate uptake and as a screening method for uptake efficiency. This method was used to compare three varieties of a high nitrogen efficiency breeding line of wheat. The originally less nitrogen efficient variety outperformed the actually sold cultivar in nitrate uptake rate.