Abstract
Immunogold labeling was used to study the time of appearance and distribution of a nodule-specific form of uricase (EC 1.7.3.3) in developing nodules of soybean (Glycine max (L.) Merr.) inoculated with Bradyrhizobium japonicum. The enzyme was detected in thin sections of tissue embedded in either L R White acrylic resin or Spurr's epoxy resin, by employing a polyclonal antibody preparation active against a subunit of soybean nodule uricase. Antigenicity was better preserved in L R White resin, but ultrastructure was better maintained in Spurr's. Uricase was first detectable with protein A-gold in young, developing peroxisomes in uninfected cells, coincident with the release of Bradyrhizobium bacteroids from infection threads in adjacent infected cells. As the peroxisomes enlarged, labeling of the dense peroxisomal matrix increased. Gold particles were never observed over the paracrystalline inclusions of peroxisomes, however. Despite a close association between enlarging peroxisomes and tubular endoplasmic reticulum, uricase was not detectable in the latter. In mature nodules, labeling of uricase was limited to the large peroxisomes in uninfected cells. Small peroxisome-like bodies present in infected cells did not become labeled.
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Abbreviations
- BSA:
-
bovine serum albumin
- Da:
-
dalton
- ER:
-
endoplasmic reticulum
- IgG:
-
immunoglobulin G
References
Atkins, C.A., Shelp, B.J., Storer, P.J., Pate, J.S. (1984) Nitrogen nutrition and the development of biochemical functions associated with nitrogen fixation and ammonia assimilation of nodules on cowpea seedlings. Planta 162, 327–333
Beevers, H. (1979) Microbodies in higher plants. Annu. Rev. Plant Physiol. 30, 159–197
Bendayan, M., Zollinger, M. (1983) Ultrastructural localization of antigenic sites on osmium-fixed tissues applying the protein A-gold technique. J. Histochem. Cytochem. 31, 101–109
Bergmann, H., Preddie, E., Verma, D.P.S. (1983) Nodulin-35: a subunit of specific uricase (uricase II) induced and localized in the uninfected cells of soybean nodules. EMBO J. 2, 2333–2339
Bisseling, T., Been, C., Klugkist, J., Van Kammen, A., Nadler, K. (1983) Nodule-specific host proteins in effective and ineffective root nodules of Pisum sativum. EMBO J. 2, 961–966
Bisseling, T., Moen, A.A., Van den Bos, R.C., Van Kammen, A. (1980) The sequence of appearance of leghaemoglobin and nitrogenase components I and II in root nodules of Pisum sativum. J. Gen. Microbiol. 118, 377–381
Brewin, N.J., Robertson, J.G., Wood, E.A., Wells, B., Larkins, A.P., Galfre, G., Butcher, G.W. (1985) Monoclonal antibodies to antigens in the peribacteroid membrane from Rhizobium-induced root nodules of pea cross-react with plasma membranes and Golgi bodies. EMBO J. 4, 605–611
Craig, S., Goodchild, D.J. (1984) Periodate-acid treatment of sections permits on-grid immunogold localization of pea seed vicilin in ER and golgi. Protoplasma 122, 35–44
Craig, S., Miller, C. (1984) L R White resin and improved on-grid immunogold detection of vicilin, a pea storage protein. Cell Biol. Int. Rep. 8, 879–886
Doman, D.C., Trelease, R.N. (1985) Protein A-gold immunocytochemistry of isocitrate lyase in cotton seeds. Protoplasma 124, 157–167
Frederick, S.E., Newcomb, E.H. (1969) Cytochemical localization of catalase in leaf microbodies (peroxisomes). J. Cell Biol. 43, 343–353
Fuller, F., Verma, D.P.S. (1984) Appearance and accumulation of nodulin in mRNAs and their transcripts and their relationships to effectiveness of root nodules. Plant Mol. Biol. 3, 21–28
Goldman, B.M., Blobel, G. (1978) Biogenesis of peroxisomes: Intracellular site of synthesis of catalase and uricase. Proc. Natl. Acad. Sci. USA 75, 5066–5070
Hanks, J.F., Schubert, K., Tolbert, N.E. (1983) Isolation and characterization of infected and uninfected cells from soybean nodules. Plant Physiol. 71, 869–873
Hanks, J.F., Tolbert, N.E., Schubert, K.R. (1981) Localization of enzymes of ureide biosynthesis in peroxisomes and microsomes of nodules. Plant Physiol. 68, 65–69
Herridge, D.F., Atkins, C.A., Pate, J.S., Rainbird, R.M. (1978) Allantoin and allantoic acid in the nitrogen economy of the cowpea. Plant Physiol. 62, 495–498
Hoagland, D.R., Snyder, W.C. (1933) Nutrition of strawberry plant under controlled conditions. Proc. Am. Soc. Hort. Sci. 30, 288–294
Huang, A.H.C., Trelease, R.N., Moore, T.S. (1983) Plant peroxisomes. Academic Press, New York London
Lazarow, P.B. (1981) Functions and biogenesis of peroxisomes, 1980. In: International cell biology 1980–1981, pp. 633–639, Schweiger, H.G., ed. Springer, Berlin Heidelberg New York
Legocki, R.P., Verma, D.P.S. (1979) A nodule-specific plant protein (Nodulin-35) from soybean. Science 205, 190–193
Legocki, R.P., Verma, D.P.S. (1980) Identification of “nodule-specific” host proteins (nodulins) involved in the development of Rhizobium-legume symbiosis. Cell 20, 153–163
Marks, I., Sprent, J.I. (1974) The localization of enzymes in fixed sections of soybean root nodules by electron microscopy. J. Cell Sci. 16, 623–637
Newcomb, E.H., Tandon, S.R. (1981) Uninfected cells of soybean root nodules: ultrastructure suggests key role in ureide production. Science 212, 1394–1396
Newcomb, E.H., Tandon, S.R., Kowal, R.R. (1985) Ultrastructural specialization for ureide production in uninfected cells of soybean root nodules. Protoplasma 125, 1–12
Nguyen, T., Zelechowska, M., Foster, V., Bergmann, H., Verma, D.P.S. (1985) Primary structure of the soybean nodulin-35 gene encoding uricase II localized in the peroxisomes of uninfected cells of nodules. Proc. Natl. Acad. Sci. USA 82, 5040–5044
Pate, J.S. (1976) Transport in symbiotic systems fixing nitrogen. In: Encyclopedia of plant physiology, N.S. vol. 2: Transport in plants IIB. Tissues and organs, pp. 278–306, Lüttge, U., Pitman, M.G., eds. Springer, Berlin Heidelberg New York
Reynolds, E.S. (1963) The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J. Cell Biol. 17, 208–213
Reynolds, P.H.S., Boland, M.J., Blevins, D.G., Schubert, K.R., Randall, D.D. (1982) Enzymes of amide and ureide biogenesis in developing soybean nodules. Plant Physiol. 69, 1334–1338
Roth, J. (1983) The protein A-gold (pAg) technique — A qualitative and quantitative approach for antigen localization on thin sections. In: Techniques in immunocytochemistry, vol. 1, pp. 107–134, Bullock, G.R., Petrusz, P., eds. Academic Press, London New York
Roth, J., Bendayan, M., Orci, L. (1978) Ultrastructural localization of intracellular antigens by the use of protein A-gold complex. J. Histochem. Cytochem. 26, 1074–1081
Schubert, K. (1981) Enzymes of purine biosynthesis and catabolism in Glycine max. I. Comparison of activities with N2 fixation and composition of xylen exudate during nodule development. Plant Physiol. 68, 1115–1122
Schubert, K., Boland, M.J. (1984) The cellular and intracellular organization of the reactions of ureide biogenesis and nodules of tropical legumes. In: Advances in nitrogen fixation research, pp. 445–452, Veeger, C., Newton, W.E., eds. Nijhoff/Junk, The Hague Boston New York
Shelp, B.J., Atkins, C.A., Storer, P.J., Canvin, D.T. (1983) Cellular and subcellular organization of pathways of ammonia assimilation and ureide synthesis in nodules of cowpea (Vigna unguiculata (L.) Walp). Arch. Biochem. Biophys. 224, 429–441
Spurr, A.R. (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastruct. Res. 26, 31–43
Titus, D.E., Becker, W.M. (1985) Investigation of the glyoxysome-peroxisome transition in germinating cucumber cotyledons using double-label immunoelectron microscopy. J. Cell Biol. 101, 1288–1299
Verma, D.P.S., Ball, S., Guérin, C., Wanamaker, L. (1979) Leghemoglobin biosynthesis in soybean root nodules. Characterization of the nascent and released peptides and the relative rate of synthesis of the major leghemoglobins. Biochemistry 18, 476–483
Verma, D.P.S., Haugland, R., Brisson, N., Legocki, R., Lacroix, L. (1981) Regulation of the expression of leghaemoglobin genes in effective and ineffective root nodules of soybean. Biochim. Biophys. Acta 653, 98–107
Verma, D.P.S., Lee, J., Fuller, F., Bergmann, H. (1984) Leghemoglobin and nodulin genes: Two major groups of host genes involved in symbiotic nitrogen fixation. In: Advances in nitrogen fixation research, pp. 557–564, Veeger, C., Newton, W.E., eds. Nijhoff/Junk, The Hague Boston New York
Weibel, E.R. (1979) Stereological methods, vol. 1. Academic Press, New York London
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Van den Bosch, K.A., Newcomb, E.H. Immunogold localization of nodule-specific uricase in developing soybean root nodules. Planta 167, 425–436 (1986). https://doi.org/10.1007/BF00391217
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DOI: https://doi.org/10.1007/BF00391217