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Temporal and Spatial Order of Events During the Induction of Cortical Cell Divisions in White Clover by Rhizobium leguminosarum bv. trifolii Inoculation or Localized Cytokinin Addition

June 2000 , Volume 13 , Number  6
Pages  617 - 628

Ulrike Mathesius , 1 Celine Charon , 2 Barry G. Rolfe , 1 Adam Kondorosi , 2 and Martin Crespi 2

1Plant Microbe Interactions Group, Research School of Biological Sciences, Australian National University (ANU), GPO Box 475, Canberra ACT 2601, Australia; 2Institute des Sciences Végétales, Centre National de la Recherche Scientifique, 1 Avenue de la Terrasse, F-91198, Gif sur Yvette Cedex, France


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Accepted 7 March 2000.

We examined the timing and location of several early root responses to Rhizobium leguminosarum bv. trifolii infection, compared with a localized addition of cytokinin in white clover, to study the role of cytokinin in early signaling during nodule initiation. Induction of ENOD40 expression by either rhizobia or cytokinin was similar in timing and location and occurred in nodule progenitor cells in the inner cortex. Inoculation of rhizobia in the mature root failed to induce ENOD40 expression and cortical cell divisions (ccd). Nitrate addition at levels repressing nodule formation inhibited ENOD40 induction by rhizobia but not by cytokinin. ENOD40 expression was not induced by auxin, an auxin transport inhibitor, or an ethylene precursor. In contrast to rhizobia, cytokinin addition was not sufficient to induce a modulation of the auxin flow, the induction of specific chalcone synthase genes, and the accumulation of fluorescent compounds associated with nodule initiation. However, cytokinin addition was sufficient for the localized induction of auxin-induced GH3 gene expression and the initiation of ccd. Our results suggest that rhizobia induce cytokinin-mediated events in parallel to changes in auxin-related responses during nodule initiation and support a role of ENOD40 in regulating ccd. We propose a model for the interactions of cytokinin with auxin, ENOD40, flavonoids, and nitrate during nodulation.


Additional keywords: auxin transport inhibition, fluorescence, GH3:gusA, signal transduction.

© 2000 The American Phytopathological Society