Effect of nitrate, ammonium, light and a plastidic factor on the appearance of multiple forms of nitrate reductase in mustard (Sinapis alba L.) cotyledons

Abstract

In mustard (Sinapis alba L.) cotyledons, four different forms of nitrate reductase (NR) can be separated by anion-exchange chromatography. Two of these forms (NR1 and NR2) appear in the presence of NO 3 (-) while the other two (NR3 and NR4) appear as a response to the application of NH 4 (+) as the sole nitrogen source. In the presence of NH4NO3, NR3 appears to be superimposed on nitrate-induced NR1 and NR2 while the NH 4 (+) -induced appearance of NR4 is totally abolished in the presence of equimolar amounts of NO 3 (-) . The appearance of NR1, NR2 and NR3 is strongly stimulated by red light pulses which operate via the far-red-absorbing form of phytochrome (Pfr), whereas the appearance of NR4 requires continuous light (likewise operating through pytochrome). Continuous red light is more effective in this case than continuous far-red light. Analysis of the data shows that the mode of action of phytochrome (Pfr) is the same in the case of the appearances of NR1 and NR2, whereas it is quantitatively different in the case of NR3 and totally different in that of NR4. A 'plastidic factor' has previously been postulated to be obligatorily involved in the transcriptional control of nuclear genes encoding for proteins destined for the chloroplast. Photooxidative damage of the plastid is postulated to destroy the ability of the organelle to produce this signal. If the plastids are damaged by photooxidation, the action of nitrate and phytochrome on the appearance of NR is abolished. The plant cell regulates the appearance of nitrate-induced NR, which is cytosolic, as if it were a plastidic protein. The appearance of NR3 depends on the plastidic factor in principally the same way as that of NR1 and NR2 whereas NR4 is totally independent of the plastidic factor. The data document particular kinds of interaction between controlling factors (light, nitrate, ammonium, plastidic factor) which affect gene expression in plants. These intricacies of regulation have so far not been considered in molecular studies on NR-gene expression.