Expression of pH-sensitive green fluorescent protein in Arabidopsis thaliana

• Published in: Plant, cell and environment cell and environment, 2001-05, Vol.24 (5), p.557-563
• Main Authors: Moseyko, N., Feldman, L. J.
• Format: Article
• Language: English
• Subjects: Arabidopsis - chemistry; Arabidopsis - cytology; Arabidopsis - genetics; Biological and medical sciences; Cell physiology; Cytoplasm - chemistry; fluorescence ratio imaging; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Plant; Genes, Plant; Genes, Reporter - genetics; green fluorescent protein (GFP); Green Fluorescent Proteins; Hydrogen-Ion Concentration; Life Sciences (General); Luminescent Proteins - genetics; Microscopy, Fluorescence; Nicotiana - genetics; Plant physiology and development; Plant Roots - chemistry; Plant Roots - cytology; Plant Roots - genetics; Plants, Genetically Modified; Plasma membrane and permeation; Plasmids - genetics; proton fluxes; Protons
• ISSN: 0140-7791; 1365-3040
• DOI: 10.1046/j.1365-3040.2001.00703.x

This is the first report on using green fluorescent protein (GFP) as a pH reporter in plants. Proton fluxes and pH regulation play important roles in plant cellular activity and therefore, it would be extremely helpful to have a plant gene reporter system for rapid, non-invasive visualization of intracellular pH changes. In order to develop such a system, we constructed three vectors for transient and stable transformation of plant cells with a pH-sensitive derivative of green fluorescent protein. Using these vectors, transgenic Arabidopsis thaliana and tobacco plants were produced. Here the application of pH-sensitive GFP technology in plants is described and, for the first time, the visualization of pH gradients between different developmental compartments in intact whole-root tissues of A. thaliana is reported. The utility of pH-sensitive GFP in revealing rapid, environmentally induced changes in cytoplasmic pH in roots is also demonstrated.