Data di Pubblicazione:
2008
Citazione:
Isolation of a strong Arabidopsis guard cell promoter and its potential as a research tool / Y. Yang, A. Costa, N. Leonhardt, R.S. Siegel, J.I. Schroeder. - In: PLANT METHODS. - ISSN 1746-4811. - 4:1(2008 Feb), p. 6.art. n°6. [10.1186/1746-4811-4-6]
Abstract:
Background: A common limitation in guard cell signaling research is that it is difficult to obtain
consistent high expression of transgenes of interest in Arabidopsis guard cells using known guard
cell promoters or the constitutive 35S cauliflower mosaic virus promoter. An additional drawback
of the 35S promoter is that ectopically expressing a gene throughout the organism could cause
pleiotropic effects. To improve available methods for targeted gene expression in guard cells, we
isolated strong guard cell promoter candidates based on new guard cell-specific microarray
analyses of 23,000 genes that are made available together with this report.
Results: A promoter, pGC1(At1g22690), drove strong and relatively specific reporter gene
expression in guard cells including GUS (beta-glucuronidase) and yellow cameleon YC3.60 (GFPbased
calcium FRET reporter). Reporter gene expression was weaker in immature guard cells. The
expression of YC3.60 was sufficiently strong to image intracellular Ca2+ dynamics in guard cells of
intact plants and resolved spontaneous calcium transients in guard cells. The GC1 promoter also
mediated strong reporter expression in clustered stomata in the stomatal development mutant toomany-
mouths (tmm). Furthermore, the same promoter::reporter constructs also drove guard cell
specific reporter expression in tobacco, illustrating the potential of this promoter as a method for
high level expression in guard cells. A serial deletion of the promoter defined a guard cell
expression promoter region. In addition, anti-sense repression using pGC1 was powerful for
reducing specific GFP gene expression in guard cells while expression in leaf epidermal cells was
not repressed, demonstrating strong cell-type preferential gene repression.
Conclusion: The pGC1 promoter described here drives strong reporter expression in guard cells
of Arabidopsis and tobacco plants. It provides a potent research tool for targeted guard cell
expression or gene silencing. It is also applicable to reduce specific gene expression in guard cells,
providing a method for circumvention of limitations arising from genetic redundancy and lethality.
These advances could be very useful for manipulating signaling pathways in guard cells and modifying
plant performance under stress conditions. In addition, new guard cell and mesophyll cell-specific
23,000 gene microarray data are made publicly available here.
Tipologia IRIS:
01 - Articolo su periodico
Elenco autori:
Y. Yang, A. Costa, N. Leonhardt, R.S. Siegel, J.I. Schroeder
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