A PAL1 gene promoter-green fluorescent protein reporter system to analyse defence responses in live cells of Arabidopsis thaliana

Rookes, James and Cahill, David 2003, A PAL1 gene promoter-green fluorescent protein reporter system to analyse defence responses in live cells of Arabidopsis thaliana, European journal of plant pathology, vol. 109, no. 1, pp. 83-94.

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Title A PAL1 gene promoter-green fluorescent protein reporter system to analyse defence responses in live cells of Arabidopsis thaliana
Author(s) Rookes, James
Cahill, David
Journal name European journal of plant pathology
Volume number 109
Issue number 1
Start page 83
End page 94
Publisher Kluwer Academic Publishers
Place of publication Dordrecht, Netherlands
Publication date 2003-01
ISSN 0929-1873
1573-8469
Keyword(s) peronospora parasitica
phenylalanine ammonia
lyase
pseudomonas syringae pv. tomato
spectrofluorometry
Summary Arabidopsis thaliana ecotype Columbia-0 was transformed with a green fluorescent protein (GFP) gene under control of a phenylalanine ammonia-lyase (PAL) promoter. PAL is a key enzyme of the phenylpropanoid pathway and is induced to high levels during plant stress. Constitutive expression of PAL1 promoter-controlled GFP occurred in vascular tissues within stems, leaves and roots and in developing flowers. PAL1 promoter–GFP expression was examined in leaves of transgenic plants subjected to an abiotic elicitor, mechanical wounding or to inoculation with the pathogens Pseudomonas syringae pv. tomato or Peronospora parasitica. Wounding of leaves and treatment with an abiotic elicitor and compatible interactions produced low to moderate levels of GFP. However, in incompatible interactions there were high levels of GFP produced. In incompatible interactions, the intensity of GFP fluorescence was similar to that produced in transgenic plants expressing GFP driven by the CaMV promoter. The bright green fluorescence produced in live cells and tissues was readily visualised using conventional fluorescence microscopy and was quantified using spectroflourometry. This is the first report of the use of GFP as a reporter of defence gene activation against pathogens. It has several advantages over other reporter genes including real time analysis of gene expression and visualisation of defence gene activation in a non-invasive manner.
Language eng
Field of Research 060499 Genetics not elsewhere classified
Socio Economic Objective 970106 Expanding Knowledge in the Biological Sciences
HERDC Research category C1 Refereed article in a scholarly journal
Copyright notice ©2003, Kluwer Academic Publishers
Persistent URL http://hdl.handle.net/10536/DRO/DU:30001989

Document type: Journal Article
Collection: School of Biological and Chemical Sciences
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