Abstract
Corn seedlings (Zea mays cv W64A × W182E) were grown hydroponically, in the presence or absence of NO3−, with or without light and with NH4Cl as the only N source. In agreement with earlier results nitrate reductase (NR) activity was found only in plants treated with both light and NO3−. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by transfer of the proteins to nitrocellulose paper and reaction with antibodies prepared against a pure NR showed that crude extracts prepared from light-grown plants had a polypeptide of approximately 116 kilodaltons (the subunit size for NR) when NO3− was present in the growth medium. Crude extracts from plants grown in the dark did not have the 116 kilodalton polypeptide, although smaller polypeptides, which reacted with NR-immunoglobulin G, were sometimes found at the gel front. When seedlings were grown on Kimpack paper or well washed sand, NR activity was again found only when the seedlings were exposed to light and NO3−. Under these conditions, however, a protein of about 116 kilodaltons, which reacted with the NR antibody was present in light-grown plants whether NO3− was added to the system or not. The NR antibody cross-reacting protein was also seen in hydroponically grown plants when NH4Cl− was the only added form of nitrogen. These results indicate that the induction of an inactive NR-protein precursor in corn is mediated either by extremely low levels of NO3− or by some other unidentified factor, and that higher levels of NO3− are necessary for converting the inactive NR cross-reacting protein to a form of the enzyme capable of reducing NO3− to NO2−.
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