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First published online October 28, 2005; 10.1104/pp.105.068262 Plant Physiology 139:1421-1432 (2005) © 2005 American Society of Plant Biologists An Egg Apparatus-Specific Enhancer of Arabidopsis, Identified by Enhancer Detection1CAMBIA, Canberra, Australian Capital Territory 2601, Australia (W.Y., R.A.J., E.H.); Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724 (J.M.M., W.B.G., U.G.); and Institute of Plant Biology, University of Zürich, CH8008 Zurich, Switzerland (U.G.)
Despite a central role in angiosperm reproduction, few gametophyte-specific genes and promoters have been isolated, particularly for the inaccessible female gametophyte (embryo sac). Using the Ds-based enhancer-detector line ET253, we have cloned an egg apparatus-specific enhancer (EASE) from Arabidopsis (Arabidopsis thaliana). The genomic region flanking the Ds insertion site was further analyzed by examining its capability to control gusA and GFP reporter gene expression in the embryo sac in a transgenic context. Through analysis of a 5' and 3' deletion series in transgenic Arabidopsis, the sequence responsible for egg apparatus-specific expression was delineated to 77 bp. Our data showed that this enhancer is unique in the Arabidopsis genome, is conserved among different accessions, and shows an unusual pattern of sequence variation. This EASE works independently of position and orientation in Arabidopsis but is probably not associated with any nearby gene, suggesting either that it acts over a large distance or that a cryptic element was detected. Embryo-specific ablation in Arabidopsis was achieved by transactivation of a diphtheria toxin gene under the control of the EASE. The potential application of the EASE element and similar control elements as part of an open-source biotechnology toolkit for apomixis is discussed.
1 This work was supported in part by the Rockefeller Foundation. W.Y. was supported by a Ph.D. fellowship from the Rockefeller Foundation. U.G. acknowledges the support of the Cold Spring Harbor Laboratory President's Council, the European Molecular Biology Organization, and the Human Frontiers Science Program. 2 Present address: Diversity Arrays Technology Pty. Ltd., P.O. Box E4008, Canberra, ACT 2604, Australia. 3 Present address: Institute of Plant Sciences, University of Berne, CH3013 Berne, Switzerland. The authors 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) are: Wei Yang (wei{at}cambia.org) and Richard A. Jefferson (r.jefferson{at}cambia.org). Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.105.068262. * Corresponding author; e-mail raj{at}cambia.org; fax 61262464533. Received July 10, 2005; returned for revision September 20, 2005; accepted September 21, 2005. Related articles in Plant Physiol.:
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