Monitoring Phytoremediation Process Using the Green Fluorescent Protein

The removal of pollutant metal ions from water using phytoremediation must be monitored to determine the effectiveness of the process. In particular, a rapid, low cost, in situ method for monitoring metal concentrations in both polluted water and plants is needed to provide the operator of a continuous phytoremediation water purification system with the information required to rotate and replace plants as required, in order to maximize metal ion removal. This project will develop ¿indicator¿ plants, utilizing the Green Fluorescent Protein (GFP), which, when expressed in either eukaryotic or prokaryotic cells and illuminated with UV light, yields a bright green fluorescence that can be used as a rapid, simple assay for the presence of the protein. If the GFP gene were fused to a metal-regulated-promoter (MRP), the resulting plants would express GFP only in the presence of certain metal ions. These plants could then be used to monitor the concentration of pollutant metal ions in water and possibly soil. The same plants could also act as indicators of water purity, providing in situ monitoring of the purity of the water outflow from a phytoremediation water treatment system. Phase I will clone the MRP from Indian mustard, develop a plant transformation vector containing MRP-GFP constructs, stablize the Agrobacterium-mediated germ-line transformation of Arabidopsis with the MRP-GFP constructs, and characterize the metal-regulated expression of GFP in Arabidopsis transformed with the MRP-GFP constructs