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Journal of Experimental Botany, Vol. 53, No. 369, pp. 621-629, April 1, 2002
© 2002 Oxford University Press

Original Papers

Local expression of the ipt gene in transgenic tobacco (Nicotiana tabacum L. cv. SR1) axillary buds establishes a role for cytokinins in tuberization and sink formation

Anne Guivarc'h1,4, Jacques Rembur1, Marc Goetz2, Thomas Roitsch2, Michèle Noin1, Thomas Schmülling3 and Dominique Chriqui1

1 University Pierre and Marie Curie, Laboratory CEMV, Bât. N2, 4, place Jussieu, F-75252 Paris Cedex 05, France
2 Universität Regensburg, Lehrstuhl fuer Zellbiologie und Pflanzenphysiologie, Universitaetsstrasse 31, D-93053 Regensburg, Germany
3 Universität Tübingen, ZMBP, Allgemeine Genetik, Auf der Morgenstelle 28, D-72076 Tübingen, Germany

The developmental characteristics of a transgenic tobacco line (BIK62) expressing the ipt cytokinin-biosynthetic gene under the control of a tagged promoter were analysed. In situ hybridization and cytokinin immunocytochemistry revealed that the ipt gene was mainly expressed in the axillary buds after the floral transition. The ipt-expressing axillary buds presented morphological alterations such as short and narrow scale-leaflets, and swollen internodes filled with starch grains, giving rise to short and tuberized lateral branches. In addition, the modification of the endogenous cytokinin balance in the axillary meristems resulted in a fast rate of leaf initiation and cytokinins accumulated mostly in the lateral zones of the reactivated axillary meristems, suggesting a role in leaf organogenesis. Cell cycle analysis revealed that the reactivated axillary meristems were characterized by predominant S+G2 nuclei. Terminal internodes displayed low levels of hexose and sucrose concomitant with starch accumulation. Extracellular invertases (EC 3.1.26) were also present in higher amounts in the tuberizing internodes compared to the axillary buds of wild-type tobacco. These results underline the role of cytokinins in cell cycle regulation and in the creation of a sink–source effect. They also provide new information about cytokinin involvement in the process of tuberization and their overproduction in axillary buds giving rise to tuberized lateral branches in a naturally non-tuberizing species.

Key words: Apical dominance, axillary bud, cytokinins, ipt gene, tuberization.


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