Phytochrome mediated induction of nitrate reductase activity in etiolated maize leaves

The effects of red and far-red light on the enhancement of in vitro nitrate reductase activity and on nitrate accumulation in etiolated excised maize leaves were examined. Illumination for 5 min with red light followed by a 4-h dark period caused a marked increase in nitrate reductase activity, whereas a 5-min illumination with far-red light had no effect on the enzyme activity. The effect of red light was completely reversed by a subsequent illumination with the same period of far-red light. Continuous far-red light also enhanced nitrate reductase activity. Both photoreversibility by red and far-red light and the operation of high intensity reaction under continuous far-red light indicated that the induction of nitrate reductase was mediated by phytochrome. Though nitrate accumulation was slightly enhanced by red and continuous far-red light treatments by 17% and 26% respectively, this is unlikely to account for the entire increase of nitrate reductase activity. The far-red light treatments given in water, to leaves preincubated in nitrate, enhanced nitrate reductase activity considerably over the dark control. The presence of a lag phase and inhibition of increase in enzyme activity under continuous far-red light-by tungstate and inhibitors of RNA synthesis and protein synthesis-rules out the possibility of activation of nitrate reductase and suggests de novo synthesis of the enzyme affected by phytochrome.     

Effect of Ammonium, Sucrose and Light on the Regulation of Nitrate Reductase Level in Pisum sativum

In shoot apices of 7-day-old dark-grown peas the addition of ammonium along with the inducer nitrate resulted in a more than two-fold increase in nitrate reductase activity. Individual amino acids, amides and amino-acid mixture could not replace the ammonium effect. Ammonium also stimulated NADH-glutamate dehydrogenase but not glucose-6-phosphate dehydrogenase. Sucrose caused a marked stimulation of nitrate reductase induction and showed synergistic effect with light. In presence of cordycepin and cycloheximide, induction of nitrate reductase was inhibited more if ammonium or sucrose was supplied along with the inducer. With actinomycin D, α-amanitin or chloramphenicol, no differential inhibition took place in presence of ammonium. The inhibition of enzyme activity by chloramphenicol and 3-(3,4-dichlorophenyl)-l,dimethyl urea was completely relieved by sucrose. Incorporation of ¹⁴C-lysine was markedly stimulated by sucrose, but was not affected by ammonium. The effect of sucrose and light on ¹⁴C-lysine incorporation was additive. Cordycepin and cycloheximide did not have any differential effect on ¹⁴C-lysine incorporation in the presence of ammonium as well as sucrose. The inhibition of ¹⁴C-lysine incorporation caused by chloramphenicol was relieved by sucrose. Sucrose also caused a marked increase in ³H-uridine incorporation but ammonium had no effect. Actinomycin D and cordycepin blocked the sucrose dependent increase in ³H-uridine incorporation. The results suggest that ammonium mediated stimulation may depend on a regulatory protein(s) synthesized in response to ammonium, whereas sucrose acts mainly by an overall increase in RNA and protein synthesis. The effect of light does not seem to be dependent on photosynthetic light reactions.     

Development of Pollen Embryoids in Anther Cultures of Datura innoxia: II. EFFECTS OF GROWTH HORMONES

In anther cultures of Datura innoxia the addition of growthhormones, such as auxins, gibberellins, and cytokinins, to theculture medium enhanced the production of pollen embryoids.Cytokinins appeared to be the most effective and, among thefour cytokinins tested, zeatin and kinetin gave the best results.Generally speaking, combinations of hormones did not improvethe response over that of an individual hormone. The numberof embryoids per anther varied in the same medium and did notstrictly correlate with the percentage of responding anthers.     

The Effects of {delta}-Aminolevulinic Acid and Inhibitors of RNA and Protein Synthesis on Phytochrome Mediated Chlorophyll Accumulation in Sorghum bicolor

In 5-d-old etiolated seedlings of Sorghum bicolor, 12 h of darknessafter 5 min in red light eliminated a lag before the accumulationof chlorophylls in subsequent continuous white light. Increasingthe dark period to 24 h and 36 h, increased the rate of chlorophyllaccumulation in the later stages of greening. Exogenous -aminolevulinicacid neither completely removed the lag, nor increased the rateof chlorophyll accumulation. Cycloheximide (25 µg ml^–1)and 6-methyl purine (5.0 µg ml^–1), given continuouslyor only until the 12 h dark period following the red light irradiation,restored the lag and decreased the rate of chlorophyll accumulation.D-threo-chloramphenicol (400µg ml^–1) also decreasedthe rate of chlorophyll accumulation but did not restore thelag. Addition of these inhibitors even 12 h after red lightirradiation decreased the rate of chlorophyll accumulation.Rifampicin (Rifamycin SV, 400 µg ml^–1) did not havesuch effects. Key words: Chlorophylls, Phytochrome, -Aminolevulinic acid, Sorghum bicolor     

Stimulation of Nitrate Reductase by Light and Ammonium in Spirodela oligorrhiza

Light-enhanced nitrate reductase (NR) activity was 8 times greaterthan the dark control. Exogenous application of sucrose, glucoseand fructose increased the induction of NR in the light as wellas in the dark, whereas glycolate had no effect. DCMU [3-(3,4-dichlorophenyl)-1, 1-dimethyl urea] completely inhibited thedevelopment of NR in light. Sucrose, when added with DCMU, reversedthis inhibitory effect NR in vivo was more stable in light thanin darkness, the half-lives being 9.6 h and 6.4 h, respectively.The addition of sucrose did not change the half-life of NR ineither light or darkness. Ammonium, the end product of the inorganicnitrogen assimilatory pathway, stimulated the NR activity whereasamino acids decreased it. Key words: Spirodela oligorrhiza, nitrate reductase, ammonium, light     

Development of Pollen Embryoids in Anther Cultures of Datura innoxia: I. GENERAL OBSERVATIONS AND EFFECTS OF PHYSICAL FACTORS

To obtain the maximal production of pollen embryoids in culturedanthers of Datura innoxia, the critical stage of anther developmentand the effect of physical factors, such as the precise modeof implantation of the anthers in the culture medium, light,temperature, and pH, were studied. In almost all media used,anthers containing uninucleate pollen were the best for initiationof embryogenesis. Variations in light and temperature also affecteddevelopment of the embryoids significantly. The percentage ofanthers producing pollen embryoids increased almost linearlywhen the temperature was raised from 22 to 30 °C. At lowertemperatures (15 to 20 °C) no embryoids were produced. Cultureskept in darkness produced embryoids, but upon transfer of culturesto the light the percentage of responding anthers increasedconsiderably.