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A PAL1 gene promoter-green fluorescent protein reporter system to analyse defence responses in live cells of Arabidopsis thaliana
journal contribution
posted on 2003-01-01, 00:00 authored by Jim RookesJim Rookes, David CahillDavid CahillArabidopsis 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.
History
Journal
European journal of plant pathologyVolume
109Issue
1Pagination
83 - 94Publisher
Kluwer Academic PublishersLocation
Dordrecht, NetherlandsPublisher DOI
ISSN
0929-1873eISSN
1573-8469Language
engPublication classification
C1 Refereed article in a scholarly journalCopyright notice
2003, Kluwer Academic PublishersUsage metrics
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Categories
Keywords
peronospora parasiticaphenylalanine ammonialyasepseudomonas syringae pv. tomatospectrofluorometryScience & TechnologyLife Sciences & BiomedicineAgronomyPlant SciencesHorticultureAgriculturephenylalanine ammonia-lyaseVITAL MARKERHYPERSENSITIVE RESPONSEHIGHER-PLANTSMOSAIC-VIRUSEXPRESSIONTRANSFORMATIONINFECTIONGFPTOBACCO