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GENETICS, GENOMICS, AND MOLECULAR EVOLUTION

Novel Markers of Xylogenesis in Zinnia Are Differentially Regulated by Auxin and Cytokinin^1,[W]

Unité Mixte de Recherche, Centre National de la Recherche Scientifique/Université Paul Sabatier 5546, Surfaces Cellulaires et Signalisation chez les Végétaux, Pôle de Biotechnologie Végétale, 31326 Castanet, Tolosan, France

The characterization of in vitro xylogenic cultures of zinnia (Zinnia elegans) has led to major discoveries in the understanding of xylem formation in plants. We have constructed and characterized a subtractive library from zinnia cultures enriched in genes that are specifically expressed at the onset of secondary wall deposition and tracheary element (TE) programmed cell death. This Late Xylogenesis Library (LXL) consisted of 236 nonredundant cDNAs, 77% of which encoded novel sequences in comparison with the 17,622 expressed sequence tag sequences publicly available. cDNA arrays were constructed to examine dynamic global gene expression during the course of TE formation. As a first step in dissecting auxin and cytokinin signaling during TE differentiation, macroarrays were probed with cDNAs from cells cultured in different hormonal conditions. Fifty-one percent of the LXL genes were induced by either auxin or cytokinin individually, the large majority by auxin. To determine the potential involvement of these categories of genes in TE differentiation, multiplex in situ-reverse transcription-PCR was performed on cells for two genes encoding putative cell wall proteins: Gibberellin stimulated transcript-1, induced by auxin alone, and expansin 5, induced by cytokinin alone. All transcriptionally active TEs expressed both genes, indicating that, although these genes may not be considered as specific markers for TE differentiation per se, they are nevertheless an integral part of TE differentiation program. Among the non-TE population, four different gene expression-based cell types could be distinguished. Together, these results demonstrate the underlying complexity of hormonal perception and the existence of several different cell types in in vitro TE cell cultures.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Deborah Goffner (goffner{at}scsv.ups-tlse.fr).

^[W] The online version of this article contains Web-only data.

Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.105.064337.

^* Corresponding author; e-mail goffner{at}scsv.ups-tlse.fr; fax 33–5–62–19–35–02.

Received April 15, 2005; returned for revision July 21, 2005; accepted September 2, 2005.

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