High throughput screening of signal transduction mutants with luciferase lmaging

摘要

A detailed procedure for high throughput genetic screening of hormone and environmental stress signal transduction mutants of Arabidopsis thaliana is described. The screen was carried out with mutagenized plants expressing the firefly luciferase reporter under control of a cold, osmotic stress, and absciscic acid responsive promoter. A thermoelectrically cooled CCD camera was used to detect luminescence emitted by the plants in response to stresses or ABA. Advantages of the screening procedure include high throughput, capability to identify low as well as high expression mutants and employment of a highly sensitive but affordable imaging system and software. This procedure can be used to study complex signal transduction networks in higher plants.