Abstract
Induction of nitrate reductase EC 1.6.6.1 in etiolated barley (Hordeum vulgare L., var. Proctor) required continuous illumination and showed a lag period of about three hours. During the first 16 h of illumination the ratio NADH/NAD and NADPH/NADP, taken as a measure of internal oxidation reduction potential, declined. The inhibitor DCMU applied to whole leaves at concentrations shown to inhibit the reduction of cytochrome f by Photosystem 2 light did not inhibit the induction of nitrate reductase nor did it diminish the ratio of reduced to oxidised puridine nucleotides in the early hours of greening. It was concluded that light driven electron flow was not necessary for nitrate reductase induction. Chloramphenicol gave a slight inhibition of nitrate reductase induction. Laevulinic acid was added to greening barley leaves to inhibit tetrapyrrole pigment biosynthesis and plastid development. It strongly inhibited chlorophyll synthesis and nitrate reductase induction, with relatively little effect upon Photosystem 1 and 2 activities in isolated plastids. The activities of other inducible enzymes and control enzymes were little affected by laevulinic acid. Laevulinic acid also inhibited nitrate reductase induction by added nitrate in fully-greened illuminated plants grown in nitrate-free medium and so is unlikely to be acting through inhibition of plastid development. This inhibitor lowered the level of protohaem in whole leaves and plastids of greening barley and it is postulated that it may diminish the protohaem available for the assembly of a cytochrome b component of nitrate reductase.
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Abbreviations
- DCMU:
-
3-(3:4-Dichlorophenyl)-1:1-dimethylurea
- LA:
-
laevulinic acid
References
Anderson, L.E., Avron, M.: Light modulation of enzyme activity in chloroplasts. Plant Physiol. 57, 209–213 (1976)
Arnon, D.: Copper enzymes in isolated chloroplasts. Polyphenol-oxidase in Beta vulgaris. Plant Physiol. 2, 1–15 (1949)
Beale, S.I., Castelfranco, P.A.: The biosynthesis of 5-aminolaevulinic acid in higher plants. I. Accumulation of 5-aminolaevulinic acid in greening plant tissues. Plant Physiol. 53, 291–296 (1974a)
Beale, S.I., Castelfranco, P.A.: II. Formation of 14C 5-aminolaevulinic acid from labelled precursors in greening plant tissues. Plant Physiol. 53, 297–303 (1974b)
Beevers, L., Hageman, R.H.: Nitrate reduction in higher plants. Ann. Rev. Plant Physiol. 20, 495–522 (1969)
Beevers, L., Hageman, R.H.: The role of light in nitrate metabolism in higher plants. Photophysiology. VII, 85–113. A.C. Giese, ed. New York and London: Academic Press 1972
Bradbeer, J.W.: The activities of the photosynthetic carbon cycle enzymes of greening bean leaves. New Phytol. 68, 233–245 (1969)
Butz, R.G., Jackson, W.A.: A mechanism for nitrate transport and reduction. Phytochemistry 16, 409–417 (1977)
Candela, M.C., Fisher, E.G., Hewitt, E.J.: Some factors affecting the activity of nitrate reductase in cauliflower plants grown with different nitrogen sources and molybdenum levels in sand cultures. Plant Physiol. 32, 280–288 (1957)
Castelfranco, P.A., Jones O.T.G.: Protohaem turnover and chlorophyll synthesis in greening barley tissue. Plant Physiol. 55, 485–490 (1975)
Chance, B.: Techniques for the assay of respiratory enzymes. In: Methods in Enzymol. Vol. VI. pp. 273–329, Colowick, S.P., Kaplan, N.O., eds. New York: Academic Press 1957)
Eaglesham, A.R.J., Hewitt, E.J.: The regulation of nitrate reductase activity from spinach (Spinacea oleracea) leaves by thiol compounds in the presence of adenosine-5′-diphosphate. FEBS Lett. 16, 315–318 (1971)
Ellis, R.J.: Chloroplast ribosomes: Stereospecificity of inhibition by chloramphenico. Science 163, 477–478 (1975)
Griffiths, W.T.: Some observations on chlorophyll(ide) synthesis by isolated etioplasts. Biochem. J. 146, 17–24 (1975)
Heber, U.W., Santarius, K.A.: Compartmentation and reduction of pyridine nucleotides in relation to photosynthesis. Biochim. Biophys. Acta. 109, 390–408 (1965)
Heldt, H.W., Rapley, L.: Specific transport of inorganic phosphate, 3-phosphoglycerate and dihydroxyacetonephosphate, and of dicarboxylates across the inner membrane of spinach chloroplasts. FEBS Lett. 10, 143–148 (1970)
Hewitt, E.J.: Assimilatory nitrate-nitrite reduction. Ann. Rev. Plant Physiol. 26, 73–100 (1975)
Hucklesby, D.P., Dalling, M.J., Hageman, R.H.: Some properties of two forms of nitrite reductase from corn scutellum. Planta 104, 220–233 (1972)
Jones, R.W., Sheard, R.W.: Nitrate reductase activity: Phytochrome mediation of induction in etiolated peas. Nature New Biology 238, 221–222 (1973)
Jones, R.W., Sheard, R.W.: Nitrate reductase activity of darkgrown and light exposed etiolated field peas (Pisum arvense). Canad. J. Bot. 51, 27–35 (1973)
Kemp, M.B., Haddock, B.A., Garland, P.B.: Synthesis and sidedness of membrane-bound respiratory nitrate reductase in Escherichia coli lacking cytochromes. Biochem. J. 148, 329–333 (1975)
Klein, S., Harel, E., Ne'eman, E., Katz, E., Meller, E.: Accumulation of 5-aminolaevulinic acid and its relation to chlorophyll synthesis and development of plastid structure in greening leaves. Plant Physiol. 56, 486–496 (1975)
Knypl, J.S.: Induction of nitrate reductase by chloramphenicol in detached cucumber cotyledons. Planta 114, 311–321 (1973)
Lewis, D.: Ph.D. Thesis, Bristol University, U.K. 1963
Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J.: Protein measurement with the Folin phenol reagent. J. biol. Chem. 193, 265–275 (1951)
Luck, H.: In: Methods of enzymic analysis. Bergmeyer, H.U., ed. pp. 885–897. New York-London: Academic Press 1971
Nandi, D.L., Shemin, D.: 5-aminolaevulinic acid dehydratase of Rhodopseudomonas sphaeroides. III. Mechanism of porphobilinogen synthesis. J. biol. Chem. 243, 1236–1242 (1968)
Notton, B.A., Fido, R.J., Hewitt, E.J.: The presence of functional haem in a higher plant nitrate reductase. Plant Sc. Letters 8, 165–170 (1977)
Plaut, Z., Littan, A.: Interaction between photosynthetic CO2 fixation products, and nitrate reduction in spinach and wheat leaves. Proc. 3rd Int. Cong. Phot. II. Avron, W., ed. pp. 1507–1516. Amsterdam Oxford New York, Elsevier Scientific Publishing Company 1974
Plesnicar, M., Bendall, D.S.: The photochemical activities and electron carriers of developing barley leaves. Biochem. J. 136, 803–812 (1973)
Sawhney, S.K., Naik, M.S.: Role of light in the synthesis of nitrate reductase and nitrite reductase in rice seedlings. Biochem. J. 130, 475–485 (1972)
Sawhney, S.K., Prakash, V., Naik, M.S.: Nitrate and nitrite reductase activities in induced chlorophyll mutants of barley. FEBS Lett. 22, 200–202 (1972)
Schrader, L.E., Beevers, L., Hageman, R.H.: Differential effects of chloramphenicol on the induction of nitrate and nitrite reductase in green leaf tissues. Biochem. Biophys. Res. Commun. 26, 14–17 (1967)
Shen, T.C.: Variation in the nitrate reductase of rice seedlings. Planta 108, 21–28 (1972)
Sluiters-Scholten, C.M.Th.: Effect of chloramphenicol and cycloheximide on the induction of nitrate reductase and nitrite reductase in bean leaves. Planta 113, 229–240 (1973)
Solomonson, L.P., Lorimer, G.H., Hall, R.L., Borchers, R., Bailey, J.L.: Reduced nicotinamide adenine dinucleotide-nitrate reductase of Chlorella vulgaris. J. biol. Chem. 250, 4120–4127 (1975)
Travis, R.L., Key, J.L.: Correlation between polyribosome levels and the ability to induce nitrate reductase in dark grown corn seedlings. Plant Physiol. 48, 617–620 (1971)
Williams, G.R., Novelli, G.D.: Ribosome changes following illumination of dark grown plants. Biochim. Biophys. Acta 155, 169–182 (1968)
Wray, J.L., Filner, P.: Structural and functional relationships of enzyme activities induced by nitrate in barley. Biochem. J. 119, 715–725 (1970)
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Nasrulhaq-Boyce, A., Jones, O.T.G. The light-induced development of nitrate reductase in etiolated barley shoots: An inhibitory effect of laevulinic acid. Planta 137, 77–84 (1977). https://doi.org/10.1007/BF00394439
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DOI: https://doi.org/10.1007/BF00394439