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JXB Advance Access published online on June 20, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri210
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oupjournals.org
Received September 15, 2004
Accepted April 27, 2005

RESEARCH PAPER

Changes in DNA and microtubules during loss and re-establishment of desiccation tolerance in germinating Medicago truncatula seeds

José M. R. Faria 1, Julia Buitink 2, André A. M. van Lammeren 3, and Henk W. M. Hilhorst 4*

1 Laboratory of Plant Physiology, Wageningen University, Arboretumlaan 4, 6703 BD Wageningen, The Netherlands; Departamento de Ciências Florestais, Universidade Federal de Lavras, CP 37, CEP 37200-000, Lavras, MG, Brazil
2 UMR 1191 Physiologie Moléculaire des Semences, INRA/INH/Université d'Angers, 16 bd Lavoisier, 49045 Angers, France
3 Laboratory of Plant Cell Biology, Wageningen University, Arboretumlaan 4, 6703 BD Wageningen, The Netherlands
4 Laboratory of Plant Physiology, Wageningen University, Arboretumlaan 4, 6703 BD Wageningen, The Netherlands

* To whom correspondence should be addressed.
Henk W. M. Hilhorst, E-mail: henk.hilhorst{at}wur.nl


  Abstract

Desiccation tolerance (DT) in orthodox seeds is acquired during seed development and lost upon imbibition/germination, purportedly upon the resumption of DNA synthesis in the radicle cells. In the present study, flow cytometric analyses and visualization of microtubules (MTs) in radicle cells of seedlings of Medicago truncatula showed that up to a radicle length of 2 mm, there is neither DNA synthesis nor cell division, which were first detected in radicles with a length of 3 mm. However, DT started to be lost well before the resumption of DNA synthesis, when germinating seeds were dried back. By applying an osmotic treatment with polyethylene glycol (PEG) before dehydration, it was possible to re-establish DT in seedlings with a radicle up to 2 mm long. Dehydration of seedlings with a 2 mm radicle, with or without PEG treatment, caused disassembly of MTs and appearance of tubulin granules. Subsequent pre-humidification led to an almost complete disappearance of both MTs and tubulin granules. Upon rehydration, neither MTs nor tubulin granules were detected in radicle cells of untreated seedlings, while PEG-treated seedlings were able to reconstitute the microtubular cytoskeleton and continue their normal development. Dehydration of untreated seedlings also led to an apoptotic-like DNA fragmentation in radicle cells, while in PEG-treated seedlingss DNA integrity was maintained. The results showed that for different cellular components, desiccation-tolerant seedlings may apply distinct strategies to survive dehydration, either by avoidance or further repair of the damages.

Keywords: Cell cycle; desiccation tolerance; DNA content; DNA integrity; Medicago truncatula; microtubules.
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