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

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.

Investigating strain transfer in polymer coated structures for the health monitoring of aerospace vehicles using polymer photonic waveguides

In this work, we present the concept of planar polymer photonic waveguides for the health monitoring of aerospace structures. Here a polymer layer is deposited onto the material/structure to be monitored. Within the polymer layer, waveguides are created after deposition. These waveguides can then be used as 'optical fibres' for optical fibre sensing methodologies. In investigating the use of polymer photonic waveguides the question to be answered is: does the strain in the test material transfer to the polymer layer, such that the value to be measured optically is reliable and indicative of the true strain in the test structure? To answer this question we have conducted a preliminary structural analysis with finite element analysis, utilising ANSYS. A simple aluminium cantilever was used as the test structure, and layers of polyethylene with different thicknesses were added to this. Result show that the thinner the layer of polymer, the more accurate the measured strain will be. For a 100um coating, the difference is strain was observed to be on the order of 3.3%. © 2014 IEEE.