First published online December 11, 2003; 10.1105/tpc.017962
The Plant Cell, Vol. 16, 74-87, January 2004,
www.plantcell.org ©2004, American Society of Plant Biologists
In Vivo Analysis of Cell Division, Cell Growth, and Differentiation at the Shoot Apical Meristem in Arabidopsis
Olivier Grandjeana,
Teva Vernouxb,
Patrick Laufsb,
Katia Belcrama,
Yuki Mizukamic and
Jan Traas1,b
a Institut National de la Recherche Agronomique, Laboratoire Commun de Cytologie, Institut Jean-Pierre Bourgin, 78026 Versailles Cedex, France
b Institut National de la Recherche Agronomique, Laboratoire de Biologie Cellulaire, Institut Jean-Pierre Bourgin, 78026 Versailles Cedex, France
c Plant and Microbial Biology, University of California, Berkeley, California 94720
1 To whom correspondence should be addressed. E-mail jan.traas{at}versailles.inra.fr; fax 33-1-30-83-30-99
The aerial parts of the plant are generated by groups of rapidly dividing cells called shoot apical meristems. To analyze cell behavior in these structures, we developed a technique to visualize living shoot apical meristems using the confocal microscope. This method, combined with green fluorescent protein marker lines and vital stains, allows us to follow the dynamics of cell proliferation, cell expansion, and cell differentiation at the shoot apex. Using this approach, the effects of several mitotic drugs on meristem development were studied. Oryzalin (depolymerizing microtubules) very rapidly caused cell division arrest. Nevertheless, both cell expansion and cell differentiation proceeded in the treated meristems. Interestingly, DNA synthesis was not blocked, and the meristematic cells went through several rounds of endoreduplication in the presence of the drug. We next treated the meristems with two inhibitors of DNA synthesis, aphidicolin and hydroxyurea. In this case, cell growth and, later, cell differentiation were inhibited, suggesting an important role for DNA synthesis in growth and patterning.
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