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JXB Advance Access published online on November 25, 2008
Journal of Experimental Botany, doi:10.1093/jxb/ern292
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© 2008 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details)

RESEARCH PAPER

GAL4 GFP enhancer trap lines for analysis of stomatal guard cell development and gene expression

Michael J. Gardner1, Andrew J. Baker1, Jean-Maurice Assie1, R. Scott Poethig2, Jim P. Haseloff1 and Alex A. R. Webb1,*

1Department of Plant Sciences, University of Cambridge, Downing Street, Cambridge CB2 3EA, UK
2Department of Biology, University of Pennsylvania, Philadelphia, PA, USA

* To whom correspondence should be addressed: E-mail: alex.webb{at}plantsci.cam.ac.uk

To facilitate the monitoring of guard cells during development and isolation, a population of 704 GAL4 GFP enhancer trap lines was screened and four single insert lines with guard cell GFP expression and one with developmentally-regulated guard cell GFP expression were identified. The location of the T-DNA inserts, the expression of the flanking genes, and the promoter activity of the genomic DNA upstream of the T-DNA were characterized. The results indicated that the GFP expression pattern in at least one of the lines was due to elements in the intergenic DNA immediately upstream of the T-DNA, rather than due to the activity of the promoters of genes flanking the insert, and provide evidence for the involvement of Dof elements in regulating guard cell gene expression. It is shown further that the GAL4 GFP lines can be used to track the contribution of guard cell material in vitro, and this method was used to assess the purity of guard cell samples obtained using two methods of guard cell isolation.

Key words: Arabidopsis, development, enhancer trap, GFP, guard cells, stomata, T-DNA

Received 21 July 2008; Revised 7 October 2008 Accepted 16 October 2008


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