Abstract
The two isoenzymes of NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14), previously identified in root nodules of Phaseolus vulgaris L., have both been shown to be located in root-nodule plastids. The nodule specific NADH-GOGAT II accounts for the majority of the activity in root nodules, and is present almost exclusively in the central tissue of the nodule. However about 20% of NADH-GOGAT I activity is present in the nodule cortex, at about the same specific activity as this isoenzyme is found in the central tissue. Glutamine synthetase (GS; EC 6.3.1.2) occurs predominantly as the γ polypeptide in the central tissue, whereas in the cortex, the enzyme is represented mainly by the β polypeptide. Over 90% of both GS and NADH-GOGAT activities are located in the central tissue of the nodule and GS activity exceeds NADH-GOGAT activity by about twofold in this region. Using the above information, a model for the subcellular location and stoichiometry of nitrogen metabolism in the central tissue of P. vulgaris root nodules is presented.
Similar content being viewed by others
Abbreviations
- Fd-GOGAT:
-
ferredoxin-dependent glutamate synthase
- GOGAT:
-
glutamate synthase
- GS:
-
glutamine synthetase
- NADH-GOGAT:
-
NADH-dependent glutamate synthase
- IEX-HPLC:
-
ion-exchange high-performance liquid chromatography
References
Appleby, C. (1969) Properties of leghaemoglobin in vivo and its isolation as ferrous oxyleghaemoglobin. Biochim. Biophys. Acta 188, 222–229
Awonaike, K.O. (1980) Studies on the development and properties of ammonia assimilating enzymes in Phaseolus vulgaris L. root nodules. Ph.D. thesis, Rothamsted Experimental Station, Harpenden, Hertfordshire
Awonaike, K.O., Lea, P.J., Miflin, B.J. (1981) The localisation of the enzymes of ammonia assimilation in root nodules of Phaseolus vulgaris L. Plant Sci. Lett 23, 189–195
Bennett, M.J., Cullimore, J.V. (1989) Glutamine synthetase isoenzymes of Phaseolus vulgaris L.: subunit composition in developing root nodules and plumules. Planta 179, 433–440
Bennett, M.J., Lightfoot, D.A., Cullimore, J.V. (1989) cDNA sequence and differential expression of the gene encoding the glutamine synthetase γ polypeptide of Phaseolus vulgaris L. Plant Mol. Biol. 12, 553–565
Boland, M.J., Farnden, K.J.F., Robertson, J.G. (1980) Ammonia assimilation in nitrogen-fixing legume nodules. In: Nitrogen fixation vol. II pp. 33–52, Newton, W.E., Orme-Johnston, W.H., eds. University Park Press, Baltimore
Boland, M.J., Hanks, J.F., Reynolds, P.H.S., Blevins, D.G., Tolbert, N.E., Schubert, K.R. (1982) Subcellular organisation of ureide biogenesis from glycolytic intermediates and ammonium in nitrogen fixing soybean nodules. Planta 155, 45–51
Boland, M.J., Schubert, K.R. (1983a) Biosynthesis of purines by a proplastid fraction from soybean nodules. Arch. Biochem. Biophys. 220, 179–187
Boland, M.J., Schubert, K.R. (1983b) Phosphoglycerate dehydrogenase from soybean nodules. Plant Physiol. 71, 658–661
Chen, F-L., Cullimore, J.V. (1988) Two isoenzymes of NADH-dependent glutamate synthase in root nodules of Phaseolus vulgaris L. Plant Physiol. 88, 1411–1417
Cullimore, J.V., Bennett M.J. (1988) The molecular biology and biochemistry of plant glutamine synthetase from root nodules of Phaseolus vulgaris L. and other legumes. J. Plant Physiol. 132, 387–393
Forde, B.G., Day, H.M., Turton, J.F., Shen, W-J., Cullimore, J.V., Oliver, J.E. (1989) Two glutamine synthetase genes from Phaseolus vulgaris L. display contrasting developmental and spatial patterns of expression in transgenic Lotus corniculatus plants. Plant Cell 1, 391–401
Gebhardt, C., Oliver, J.E., Forde, B.G., Saarelainen, R., Miflin, B.J. (1986) Primary structure and differential expression of glutamine synthetase genes in nodules, roots, and leaves of Phaseolus vulgaris. EMBO J. 5, 1429–1435
Gibbs, M., Turner, J.F. (1964) Enzymes of glycolysis. In: Modern methods of plant analysis, vol. 7, pp. 520–533, Linskens, H.F., Sanwal, B.D., Tracey, M.V. eds. Mihoff/Junk: The Hague
Givan, C.V., Joy, K.W., Kleczkowski, L.A. (1988) A decade of photorespiratory nitrogen cycling. TIBS 13, 433–437
Hanks, J.F., Tolbert, N.E., Schubert, K.R., (1981) Localisation of enzymes of ureide biosynthesis in peroxisomes and microsomes of nodules. Plant Physiol. 68, 56–69
Lara, M., Porta, H., Padilla, J., Folch, J., Sanchez, F. (1984) Heterogeneity of glutamine synthetase polypeptides in Phaseolus vulgaris L. Plant Physiol. 76, 1019–1023
Lightfoot, D.A., Green, N.K., Cullimore, J.V. (1988) The chloroplast-located glutamine synthetase of Phaseolus vulgaris L.: nucleotide sequence, expression in different organs and uptake into isolated chloroplasts. Plant Mol. Biol. 11, 191–202
Miflin, B.J., Cullimore, J.V. (1984) Nitrogen assimilation in the legume Rhizobium symbiosis: a joint endeavour. In: Genes involved in microbe plant interactions pp. 129–178, Verma, D.P.S., Hohn, Th., eds. Springer-Verlag, Vienna
Padilla, J.E., Campos, F., Conde, V., Lara, M., Sanchez, F. (1987) Nodule-specific glutamine synthetase is expressed before the onset of nitrogen fixation in Phaseolus vulgaris L. Plant Mol. Biol. 9, 65–74
Robertson J.G., Farnden K.J.F. (1980) Ultrastructure and metabolism of the developing legume root nodule. In: The biochemistry of plants, vol. 5, pp 66–113, Miflin, B.J., ed. Academic Press, New York
Schubert, K.R. (1986) Products of biological nitrogen fixation in higher plants: synthesis, transport, and metabolism. Annu. Rev. Plant Physiol. 37, 539–574
Shelp, B.J., Atkins, C.A. (1984) Subcellular locations of enzymes of ammonia assimilation and asparagine synthesis in root nodules of Lupinus albus L. Plant Sci. Lett. 36, 225–230
Shelp, B.J., Atkins, C.A., Storer, P.J., Canvin, D.J. (1983) Cellular and subcellular organization of pathway of ammonia assimilation and ureide synthesis in nodules of cowpea (Vigna unguiculata L. Walp.). Arch. Biochem. Biophys. 224, 429–441
Suzuki, A., Vidal, J., Nguyen, J., Gadal, P. (1984) Occurrence of ferredoxin-dependent glutamate synthase in plant cell fraction of soybean root nodules (Glycine max) FEBS Lett. 173, 204–208
VandenBosch, K.A., Newcomb, E.H. (1988) The occurrence of leghaemoglobin protein in the uninfected interstitial cells of soybean root nodules. Planta 175, 442–451
Verma, D.P.S., Fortin, M.G., Stanley, J., Mauro, V.P., Purohit, S., Morrison, N. (1986) Nodulins and nodulin genes of Glycine max. Plant Mol. Biol. 7, 51–61
Yu, J., Woo, K.C. (1988) Glutamine transport and the role of the glutamine translocator in chloroplasts. Plant Physiol. 88, 1048–1054
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Chen, FL., Cullimore, J.V. Location of two isoenzymes of NADH-dependent glutamate synthase in root nodules of Phaseolus vulgaris L.. Planta 179, 441–447 (1989). https://doi.org/10.1007/BF00397583
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00397583