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Immunogold localization of the cell-wall-matrix polysaccharides rhamnogalacturonan I and xyloglucan during cell expansion and cytokinesis inTrifolium pratense L.; implication for secretory pathways

Abstract

We have localized two cell-wall-matrix polysaccharides, the main pectic polysaccharide, rhamnogalacturonan I (RG-I), and the hemicellulose, xyloglucan (XG), in root-tip and leaf tissues of red clover (Trifolium pratense L.) using immunoelectron microscopy. Our micrographs show that in both leaf and root tissues RG-I is restricted to the middle lamella, with 80–90% of the label associated with the expanded regions of the middle lamella at the corner junctions between cells. Xyloglucan, however, is nearly exclusively located in the cellulose-microfibril-containing region of the cell wall. Thus, these cell-wall-matrix polysaccharides are present in distinct and complementary regions of the cell wall. Our results further show that during cell expansion both RG-I and XG are present within Golgi cisternae and vesicles, thus confirming that the Golgi apparatus is the main site of synthesis of the non-cellulosic cell-wall polysaccharides. No label is seen over the endoplasmic reticulum, indicating that synthesis of these complex polysaccharides is restricted to the Golgi. The distribution of RG-I and XG in root-tip cells undergoing cell division was also examined, and it was found that while XG is present in the Golgi stacks and cell plate during cytokinesis, RG-I is virtually absent from the forming cell plate.

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Abbreviations

ER:

endoplasmic reticulum

RG-I:

rhamnogalacturonan I

XG:

xyloglucan

References

  1. Albersheim, P. (1975) The walls of growing plant cells. Sci. Am.232, 80–95

  2. Albersheim, P., Mühlethaler, K., Frey-Wyssling, A. (1960) Stained pectin as seen in the electron microscope. J. Biophys. Biochem. Cytol.8, 501–506

  3. Amit, A.G., Mariuzza, R.A., Phillips, S.E.V., Poljak, R.J. (1985) Three-dimensional structure of an antigen-antibody complex at 6A resolution. Nature313, 156–158

  4. Bauer, W.D., Talmadge, K.W., Keegstra, K., Albersheim, P. (1973) The structure of plant cell walls. II. The hemicellulose of the walls of suspension-cultured sycamore cells. Plant Physiol.51, 174–187

  5. Brown, R.M. Jr., Willison, J.H.M. (1977) Golgi apparatus and plasma membrane involvement in secretion and cell surface deposition, with special emphasis on cellulose. In: International cell biology 1976–1977, pp. 267–283. Brinkley, B.R., Porter, K.R., eds. Rockefeller University Press, New York

  6. Camirand, A., Brummel, D., MacLachlan, D. (1987) Fucosylation of xyloglucan: localization of the transferase in dictyosomes of pea stem cells. Plant Physiol.84, 753–756

  7. Camirand, A., MacLachlan, D. (1986) Biosynthesis of the fucose-containing xyloglucan nonasaccharide by pea microsomal membranes. Plant Physiol.82, 379–383

  8. Conrad, P.A., Binary, L., Racusen, R.H. (1982) Rapidly-secreting, cultured oat cells serve as a model system for the study of cellular exocytosis. Characterization of cells and isolated secretory vesicles. Protoplasma112, 196–204

  9. Craig, S., Goodchild, D.J. (1984) Periodate-acid treatment of sections permits on-grid immunogold localization of pea seed vicilin in ER and Golgi. Protoplasma122, 35–44

  10. Darvill, A.G., McNeil, M., Albersheim, P., Delmer, D. (1980) The primary cell walls of flowering plants. In: The biochemistry of plants, vol. 1: The plant cell, pp. 92–162, Tolbert, N.E., ed. Academic Press, New York

  11. Dauwalder, M., Whaley, W.G. (1974) Patterns of incorporation of [3H]galactose ofZea mays root tips. J. Cell Sci.14, 11–27

  12. Delmer, D.P. (1987) Cellulose biosynthesis. Annu. Rev. Plant Physiol.38, 259–290

  13. Delmer, D.P. (1987) Cellulose biosynthesis. Annu. Rev. Plant Physiol.38, 259–290

  14. Deshpande, B.P. (1976) Observations on the fine structure of plant cell walls. I. Use of permanganate staining. Ann. Bot.40, 433–437

  15. Fahn, A. (1982) Plant anatomy, 3rd edn. Pergamon Press, New York

  16. Fernandez, D.E., Staehelin, L.A. (1985) Structural organization of ultrarapidly frozen barley aleurone cells actively involved in protein secretion. Planta165, 455–468

  17. Fowke, L.C., Pickett-Heaps, J.D. (1972) A cytochemical and autoradiographic investigation of cell wall deposition in fiber cells ofMarchantia berteroana. Protoplasma74, 19–32

  18. Frey-Wyssling, A., López-Sáez, J.F., Mühlethaler, K. (1964) Formation and development of the cell plate. J. Ultrastruc. Res.10, 422–432

  19. Gardiner, M., Chrispeels, M.J. (1975) Involvement of the Golgi apparatus in the synthesis and secretion of hydroxyprolinerich cell wall glycoproteins. Plant Physiol.55, 536–541

  20. Giddings, T.H., Brower, D., Staehelin, L.A. (1980) Visualization of particle complexes in the plasma membrane ofMicrasterias denticulata associated with the formation of cellulose microfibrils in primary and secondary cell walls. J. Cell Biol.84, 327–334

  21. Green, J.R., Northcote, D.H. (1978) The structure and function of glycoproteins synthesized during slime-polysaccharide production by membranes of the root-cap cells of maize (Zea mays). Biochem. J.170, 599–608

  22. Harris, P.J., Northcote, D.H. (1971) Polysaccharide formation in plant Golgi bodies. Biochim. Biophys. Acta237, 56–64

  23. Herth, W. (1985) Plasma membrane rosettes involved in localized thickening during xylem vessel formation ofLepidium sativum. L. Planta164, 12–21

  24. James, D.W., Jones, R.L. (1979) Intracellular localization of GDP-fucose polysaccharide fucosyl transferase in corn roots. Plant Physiol.64, 914–918

  25. Johnson, K.D., Chrispeels, M.J. (1973) Regulation of pentosan biosynthesis in barley aleurone tissue by gibberelic acid. Planta111, 353–364

  26. Jones, M.G.K., Payne, H.L. (1977) Cytokinesis inImpatiens balsamina and the effect of caffeine. Cytobios20, 79–91

  27. Labavitch, J.M., Ray, P.M. (1974) Relationship between promotion of XG metabolism and induction of elongation by IAA. Plant Physiol.54, 499–502

  28. McNeil, M., Darvill, A.G., Albersheim, P. (1980) The structure of plant cell walls. X. Rhamnogalacturonan I, a structurally complex pectic polysaccharide in the walls of suspension-cultured sycamore cells. Plant Physiol.66, 1128–1134

  29. McNeil, M., Darvill, A.G., Fry, S.C., Albersheim, P. (1984) Structure and function of the primary cell walls of plants. Annu. Rev. Biochem.53, 625–663

  30. Moore, P.J., Darvill, A.G., Albersheim, P., Staehelin, L.A. (1986) Immunogold localization of xyloglucan and rhamnogalacturonan I in the cell walls of suspension-cultured sycamore cells. Plant Physiol.82, 787–794

  31. O'Brien, T.P. (1972) The cytology of cell-wall formation in some eukaryotic cells. Bot. Rev.38, 87–118

  32. Pickett-Heaps, J.D. (1966) Incorporation of radioactivity into wheat xylem walls. Planta71, 1–14

  33. Pickett-Heaps, J.D. (1968) Further ultrastructural observations on polysaccharide localization in plant cells. J. Cell Sci3, 55–64

  34. Ray, P.M. (1980) Cooperative action of B-glucan synthetase and UDP-xylose xylosyl transferase of Golgi membranes in the synthesis of xyloglucan-like polysaccharide. Biochem. Biophys. Acta629, 431–444

  35. Ray, P.M., Eisinger, W., Robinson, D.H. (1976) Organelles involved in cell wall formation and transport in pea cells. Ber. Dtsch. Bot. Ges.89, 121–146

  36. Ray, P.M., Shininger, T.L., Ray, M.M. (1969) Isolation of glucan synthetase particles from plant cells and identification with Golgi membranes. Proc. Natl. Acad. Sci. USA64, 605–612

  37. Reynolds, E.S. (1963) The use of lead citrate at high pH as an electron-opaque stain for electron microscopy. J. Cell Biol.17, 208–212

  38. Robinson, D.G. (1985) Plant membranes. J. Wiley & Sons, New York

  39. Robinson, D.H., Eisinger, W., Ray, P.M. (1976) Dynamics of the Golgi system in the wall matrix polysaccharide synthesis and secretion by pea cells. Ber Dtsch. Bot. Ges. Bol.89, 147–161

  40. Roland, J.C., Vian, B. (1981) Use of purified endopolygalacturonase for a topochemical study of elongating cell walls at the ultrastructural level. J. Cell Sci.48, 333–343

  41. Roland, J.C., Vian, B., Reis, D. (1977) Further observations of cell wall morphogenesis and polysaccharide arrangement during plant growth. Protoplasma91, 125–141

  42. Ruel, K., Joseleau, J.P. (1984) Use of enzyme-gold complexes for the ultrastructural localization of hemicelluloses in the plant cell wall. Histochemistry81, 573–580

  43. Selvendran, R.R. (1985) Developments in the chemistry and biochemistry of pectic and hemicellulosic polymers. J. Cell Sci., Suppl.2, 51–88

  44. Slot, J.W., Geuze, H.J. (1985) A new method of preparing gold probes for multiple-labeling cytochemistry. Eur. J. Cell Biol.38, 87–93

  45. Sokal, R.R., Rohlf, F.J. (1981) Biometry, 2nd edn. W.H. Freeman, San Francisco

  46. Tandecarz, J., Lavintman, N., Cardini, C.E. (1975) Biosynthesis of starch. Formation of a glucoproteic acceptor by a potato non-sedimentable preparation. Biochim. Biophys. Acta399, 345–355

  47. Van der Woude, W.J., Morré, D.J., Bracker, C.E. (1971) Isolation and characterization of secretory vesicles in germinated pollen ofLilium longiflorum. J Cell Sci.8, 331–351

  48. Whaley, W.G., Dauwalder, M., Kephart, J.E. (1966) The Golgi apparatus and an early stage in cell plate formation. J. Ultrastruc. Res.15, 169–180

  49. Whaley, W.G., Mollenhauer, H.H. (1963) The Golgi apparatus and cell plate formation. A postulate. J. Cell Biol.17, 216–221

  50. York, W.S., Darvill, A.G., Albersheim, P. (1984) Inhibition of 2,4-dichlorophenoxyacetic acid-stimulated elongation of pea stem segments by a xyloglucan oligosaccharide. Plant Physiol.75, 295–297

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Moore, P.J., Staehelin, L.A. Immunogold localization of the cell-wall-matrix polysaccharides rhamnogalacturonan I and xyloglucan during cell expansion and cytokinesis inTrifolium pratense L.; implication for secretory pathways. Planta 174, 433–445 (1988). https://doi.org/10.1007/BF00634471

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Key words

  • Cell wall polysaccharides
  • Cell plate
  • Hemicellulose
  • Immunolocalization
  • Golgi apparatus
  • Pectin
  • Trifolium