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First published online November 6, 2003; 10.1104/pp.103.030692 Plant Physiology 133:1882-1892 (2003) © 2003 American Society of Plant Biologists Diphtheria Toxin-Mediated Cell Ablation Reveals Interregional Communication during Arabidopsis Seed Development1Developmental Genetics, Institute of Biology, Leiden University, Clusius Laboratory, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands
Fertilization of the female gametophyte in angiosperm plants initiates a process of coordinated development of embryo, endosperm, and seed coat that ensures the production of a viable seed. Mutant analysis has suggested that communication between the endosperm and the seed coat is an important determinant in this process. In addition, cell groups within the embryo, derived from the apical and from the basal cell, respectively, after zygote division, concertedly establish a functional root meristem, and cells in the apical region of the embryo are hypothesized to repress cell divisions in the basal cell-derived suspensor. The available evidence for these interregional communication events mostly relies on the analysis of mutant phenotypes in Arabidopsis. To provide independent and direct evidence for communication events, we used conditional domain-specific expression of the diphtheria toxin A chain (DTA) in developing Arabidopsis seeds. By using a collection of cell- or tissue-type-specific promoters, we show that the mGAL4:VP16/UAS two-component gene expression allows reliable spatiotemporal and conditional expression of the GFP:GUS reporter and the DTA gene in the developing embryo and endosperm. Expression of DTA in the protoderm of the embryo proper led to excessive proliferation of suspensor cells, sometimes resulting in the formation of secondary embryos. Endosperm-specific expression of DTA caused complete cessation of seed growth, followed by pattern defects in the embryo and embryo arrest. Taken together, the results presented here substantiate the evidence for and underline the importance of interregional communication in embryo and seed development and demonstrate the usefulness of conditional toxin expression as a method complementary to phenotypic analysis of developmental mutants.
1 This work was supported by the Research Council for Earth and Life Sciences (ALW), with financial aid from the Netherlands Organisation for Scientific Research (NWO). 2 Present address: Developmental Genetics, Zentrum für Molekularbiologie der Pflanzen, Universität Tübingen, Auf der Morgenstelle 3, D-72076 Tübingen, Germany. 3 Present address: Department of Cell Biology and Histology, University of Amsterdam, Academic Medical Center, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands. Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.103.030692. * Corresponding author; e-mail offringa{at}rulbim.leidenuniv.nl; fax 31-71-5274999. Received July 23, 2003; returned for revision September 15, 2003; accepted September 15, 2003. Related articles in Plant Physiol.:
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