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Plant Physiol, February 2000, Vol. 122, pp. 327-336

Cell Division and Subsequent Radicle Protrusion in Tomato Seeds Are Inhibited by Osmotic Stress But DNA Synthesis and Formation of Microtubular Cytoskeleton Are Not1

Renato D. de Castro,2 André A.M. van Lammeren, Steven P.C. Groot, Raoul J. Bino, and Henk W.M. Hilhorst*

Laboratory of Experimental Plant Morphology and Cell Biology (R.D.d.C., A.A.M.v.L.) and Laboratory of Plant Physiology (R.D.d.C., H.W.M.H.), Wageningen University, Arboretumlaan 4, 6703 BD Wageningen, The Netherlands; and Department of Reproduction Technology, Centre for Plant Breeding and Reproduction Research, CPRO, P.O. Box 16, 6700 AA Wageningen, The Netherlands (R.D.d.C., S.P.C.G., R.J.B.).

We studied cell cycle events in embryos of tomato (Lycopersicon esculentum Mill. cv Moneymaker) seeds during imbibition in water and during osmoconditioning ("priming") using both quantitative and cytological analysis of DNA synthesis and beta -tubulin accumulation. Most embryonic nuclei of dry, untreated control seeds were arrested in the G1 phase of the cell cycle. This indicated the absence of DNA synthesis (the S-phase), as confirmed by the absence of bromodeoxyuridine incorporation. In addition, beta -tubulin was not detected on western blots and microtubules were not present. During imbibition in water, DNA synthesis was activated in the radicle tip and then spread toward the cotyledons, resulting in an increase in the number of nuclei in G2. Concomitantly, beta -tubulin accumulated and was assembled into microtubular cytoskeleton networks. Both of these cell cycle events preceded cell expansion and division and subsequent growth of the radicle through the seed coat. The activation of DNA synthesis and the formation of microtubular cytoskeleton networks were also observed throughout the embryo when seeds were osmoconditioned. However, this pre-activation of the cell cycle appeared to become arrested in the G2 phase since no mitosis was observed. The pre-activation of cell cycle events in osmoconditioned seeds appeared to be correlated with enhanced germination performance during re-imbibition in water.

1 This project was supported by a doctoral fellowship from Companha de Aperfeiçoamento de Pessoal de Nível Superior to R.D.d.C. (process no. 11241/92-4), Ministry of Education, Brazil.

2 Present address: Laboratório de Sementes, Departamento de Agricultura, Universidade Federal de Lavras, Cx. Postal 37, Lavras, Minas Gerais, CEP 37200-000, Brazil.

* Corresponding author; e-mail henk.hilhorst{at}algem.pf.wau.nl; fax 31-317-484740.

© 2000 American Society of Plant Physiologists


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