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Journal of Experimental Botany, Vol. 53, No. 369, pp. 669-675, April 1, 2002
© 2002 Oxford University Press

Original Papers

Endoreduplication is not inhibited but induced by aphidicolin in cultured cells of tobacco

Anne-Hélène Quélo1, John A. Bryant2 and Jean-Pierre Verbelen1,3

1 Department of Biology, University of Antwerp U.I.A., Universiteitsplein 1, B-2610 Wilrijk, Belgium
2 Department of Biological Sciences, University of Exeter, Exeter EX4 4QG, UK

Endoreduplication is a common process in plants that allows cells to increase their DNA content. In the tobacco cell cultures studied in this work it can be induced by simple hormone deprivation. Mesophyll protoplast-derived cells cultured in the presence of NAA (auxin) and BAP (cytokinin) keep on dividing, while elongation and concomitant DNA endoreduplication are induced and maintained in a medium containing only NAA. If aphidicolin is given to the two types of culture, no effect is observed on elongating, endoreduplicating cells. However, the cells programmed for division switch to elongation and DNA endoreduplication. Thus aphidicolin, an inhibitor of the replicative DNA polymerases, {alpha} and {delta}, does not inhibit endoreduplication, and furthermore actually induces it when the mitotic cell cycle is blocked. DNA duplication and cell growth can only be completely blocked if ddTTP, an inhibitor of DNA polymerase-ß, is given together with aphidicolin. This result implies that an aphidicolin-resistant DNA polymerase, such as the repair-associated DNA polymerase-ß, can mediate DNA synthesis during endoreduplication and can substitute for polymerases-{alpha} and -{delta} when the latter are inhibited. Similar results are obtained in cultures of the BY-2 cell line by withdrawing auxins from the culture medium. In this cell line endoreduplication is induced only in a small proportion of the cells. A greater proportion of the cells are blocked in the G2 phase of the cell cycle.

Key words: Aphidicolin, DNA polymerase, endoreduplication, N. tabacum.


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