Hostname: page-component-78c5997874-lj6df Total loading time: 0 Render date: 2024-11-15T23:21:20.108Z Has data issue: false hasContentIssue false

Vicia faba germination: Synchronized cell growth and localization of nucleolin and α-tubulin.

Published online by Cambridge University Press:  22 February 2007

Yuzo Fujikura*
Affiliation:
Institute of Experimental Botany, Norman Borlaug Center for Plant Science De Montfort University, Sokolovska 6, 77200 Olomouc, Czech Republic
Jaroslav Doležel
Affiliation:
Institute of Experimental Botany, Norman Borlaug Center for Plant Science De Montfort University, Sokolovska 6, 77200 Olomouc, Czech Republic
Jarmira Cíhalíková
Affiliation:
Institute of Experimental Botany, Norman Borlaug Center for Plant Science De Montfort University, Sokolovska 6, 77200 Olomouc, Czech Republic
Laszlo Bögre
Affiliation:
Vienna Biocenter, Institute of Microbiology and Genetics, University of Vienna, Dr. Bohrgasse 9, A-1030, Austria
Erwin Heberle-Bors
Affiliation:
Vienna Biocenter, Institute of Microbiology and Genetics, University of Vienna, Dr. Bohrgasse 9, A-1030, Austria
Heribert Hirt
Affiliation:
Vienna Biocenter, Institute of Microbiology and Genetics, University of Vienna, Dr. Bohrgasse 9, A-1030, Austria
Pavla Binarová
Affiliation:
Institute of Experimental Botany, Norman Borlaug Center for Plant Science De Montfort University, Sokolovska 6, 77200 Olomouc, Czech Republic
*
*Correspondence Department of Pathophysiology, 1st Medical Faculty, Charles University, U nemocnice 5, 12853 Prague 2, Czech Republic Tel: +420.2.24.91.49.29 Fax: +420.2.24.91.28.34 Email: [email protected]

Abstract

The first cell cycle of Vicia faba L. seeds, which begins upon imbibition of dry seeds and is completed at the first mitosis after radicle protrusion, was characterised by the flow cytometry and immunodetection of nucleolin and tubulins in root tip meristems. Flow cytometry revealed highly synchronised profiles from the quiescent G1 phase to the late G2 phase, indicating uniform cell cycle progression within a root tip until the first mitosis. Using immunoblotting, nucleolin was detected in two distinct bands with the apparent molecular masses of 89 and 99 kD; the former was detected only in seeds imbibed at 4°C for 1 day whereas the latter was found at all stages examined, suggesting that the 89 kD nucleolin may be seed-specific. Unusual localization of nucleolin in cold-imbibed seeds, undetectable in half of the cells and present in nucleoplasm, was revealed by immunofluorescence microscopy. While α- and β-tubulin were detected at all stages and no significant changes in accumulation of the proteins were observed, few microtubules were detected at the beginning of germination when cells were still in the G1 phase, suggesting that microtubules may be depolymerized in the dry seeds.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1999

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Baluska, F. and Barlow, P.W. (1993) The role of the microtubular cytoskeleton in determining nuclear chromatin structure and passage of maize root cells through the cell cycle. European Journal of Cell Biology 61, 160167.Google ScholarPubMed
Belenguer, P., Caizergues-Ferrer, M., Labbé, J.C., Dorée, M. and Amalric, F. (1990) Mitosis-specific phosphorylation of nucleolin by p34cdc2 protein kinase. Molecular and Cellular Biology 10, 36073618.Google ScholarPubMed
Bewley, J.D. and Black, M. (1994) Seeds. Physiology of development and germination. (2nd edition) New York, Plenum Press.Google Scholar
Binarová, P., Cíhalíková, J. and Doleæel, J. (1993) Localization of MPM-2 recognized phosphoproteins and tubulin during cell cycle progression in synchronized Vicia faba root meristem cells. Cell Biology International 17, 847856.Google Scholar
Bino, R.J., De Vries, J.N., Kraak, H.L. and Van Pijlen, J.G. (1992) Flow cytometric determination of nuclear replication stages in tomato seeds during priming and germination. Annals of Botany 69, 231236.Google Scholar
Bino, R.J., Lanteri, S., Verhoeven, H.A. and Kraak, H.L. (1993) Flow cytometric determination of nuclear replication stages in seed tissues. Annals of Botany 72, 181187.CrossRefGoogle Scholar
Bögre, L., Jonak, C., Mink, M., Meskiene, I., Traas, J., Ha, D.T.C., Swoboda, I., Plank, C., Wagner, E., Heberle- Bors, E. and Hirt, H. (1996) Developmental and cell cycle regulation of alfalfa nucMs1, a plant homolog of the yeast Nsr1 and mammalian nucleolin. Plant Cell 8, 417428.Google Scholar
Borer, R.A., Lehner, C.F., Eppenberger, H.M. and Nigg, E.A. (1989) Major nucleolar proteins shuttle between nucleus and cytoplasm. Cell 56, 379390.CrossRefGoogle ScholarPubMed
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248254.CrossRefGoogle ScholarPubMed
Brunori, A. (1967) Relationship between DNA synthesis and water content during ripening of Vicia faba seeds. Caryologia 20, 333338.Google Scholar
Crossin, K.L. and Carney, D.H. (1981) Evidence that microtubule depolymerization early in the cell cycle is sufficient to initiate DNA synthesis. Cell 23, 6171.Google Scholar
de Castro, R.D., Zheng, X., Bergervoet, J.H.W., De Vos, C.H.R. and Bino, R.J. (1995) b-Tubulin accumulation and DNA replication in imbibing tomato seeds. Plant Physiology 109, 499504.Google Scholar
Doleæel, J. and Novak, F.J. (1984) A standard method of measurement of nuclear Feulgen-DNA content in Allium sativum L. cells by absorption cytophotometry. Mikroskopie 41, 335341.Google Scholar
Doleæel, J., Cíhalíková, J. and Lucretti, S. (1992) A highyield procedure for isolation of metaphase chromosomes from root tips of Vicia faba L. Planta 188, 9398.CrossRefGoogle Scholar
Goddard, R.H., Wick, S.M., Silflow, C.D. and Snustad, D.P. (1994) Microtubule components of the plant cell cytoskeleton. Plant Physiology 104, 16.CrossRefGoogle ScholarPubMed
Laemmli, U.K. (1970) Cleavage of structual proteins during the assembly of the head of bacteriophage T4. Nature 277, 680685.CrossRefGoogle Scholar
Lambert, A.M. and Lloyd, C.W. (1994) The higher plant microtubule cycle. pp 325341in Hyams, J.S.; Lloyd, C.W. (Eds) Microtubules. New York, Wiley Liss.Google Scholar
Lanteri, S., Kraak, H.L., De Vos, C.H.R. and Bino, R.J. (1993) Effects of osmotic preconditioning on nuclear replication activity in seeds of pepper (Capsicum annuum). Physiologia Plantarum 89, 433440.CrossRefGoogle Scholar
Lapeyre, B., Bourbon, H. and Amalric, F. (1987) Nucleolin, the major nucleolar protein of growing eukaryotic cells: An unusual protein structure revealed by the nucleotide sequence. Proceedings of National Academy of Sciences, USA 84, 14721476.Google Scholar
Linhartová, I., Dráber, P., Dráberová, E. and Viklicky, V. (1992) Immunological discrimination of b-tubulin isoforms in developing mouse brain: post-translational modification of non-class-III b-tubulins. Biochemical Journal 288, 919924.Google Scholar
Meβmer, B. and Dreyer, C. (1993) Requirements for nuclear translocation and nucleolar accumulation of nucleolin of Xenopus laevis. European Journal of Cell Biology 61, 369382.Google Scholar
Obroucheva, N.V., Antipova, O.V., Gorbova, E.N. and Kotova, L.M. (1995) Relationship between initiation of cell elongation and cell division in radicles of germinating seeds. Plant and Soil 173, 311316.Google Scholar
Pai, C.-Y., Chen, H.-K., Shen, H.-L. and Yeh, N.-H. (1995) Cell cycle-dependent alterations of a highly phosphorylated nucleolar protein p130 are associated with nucleologenesis. Journal of Cell Science 108, 19111920.CrossRefGoogle ScholarPubMed
Sgorbati, S., Sparvoli, E., Levi, M. and Chiatante, D. (1989) Bivariate cytofluorimetric analysis of nuclear protein and DNA relative to cell kinetics during germination of Pisum sativum seed. Physiologia Plantarum 75, 479484.CrossRefGoogle Scholar
Shaw, P.J. and Jordan, E.G. (1995) The nucleolus. Annual Review of Cell and Developmental Biology 11, 93121.CrossRefGoogle ScholarPubMed
Wallin, M. and Stromberg, E. (1995) Cold-stable and cold adapted microtubules. International Review of Cytology 157, 131.Google Scholar