Nitrogen assimilation and regulation of nitrate and nitrite reductases in cultured Ipomoea cells

Abstract

Ipomoea cells grown in a medium containing ammonium and nitrate preferentially used ammonium during the initial stages of growth but in the later stages assimilated nitrate rapidly regardless of the presence or absence of ammonium. Cells grown on nitrate and maintained at pH 4.8 released ammonia into the medium, whereas when they were maintained at pH 6.5 they secreted nitrite. The enzymes of nitrate assimilation were inducible by nitrate and the activities changed considerably in response to nitrogen source. The addition of ammonium 3 days after inoculation to cells highly induced for the reductases did not result in the suppression of further synthesis of the enzymes. The levels of nitrate (EC 1.6.6.1) and nitrite (EC 1.6.6.4) reductases detected in cells grown on ammonium as the nitrogen source were about 25% and 66%, respectively, of the fully induced level. The addition of nitrate to ammonium-assimilating cells resulted in a low level of induction of both reductases. Addition of nitrite had no effect. With both ammonium and nitrate initially present in the medium, the ammonium was utilized quickly but no induction of the reductases was observed for 24 h. the time interval when the assimilation of nitrate was low. This was followed by the induction of the enzymes to a higher level than the activity in cultures of similar age that had been grown continually in nitrate, supplied at the same concentration. Thus, for nitrate and nitrite reductases, repression-like effects were produced by ammonium.