RUNIONS, C. JOHN* and JIM HASELOFF. Department of Plant Science, University of Cambridge, Cambridge, UK, CB2 3EA. - Enhancer trapping: visualization of tissue-specific gene expression in living meristems.
Fluorescent proteins are increasingly being used as a cellular marker
in the study of plant development. We use enhancer trapping, a
transactivating system that results in tissue specific fluorescent
protein expression, to study development of the apical meristems of
Arabidopsis. Tissue initiation and proliferation can be
documented via confocal microscopy and then imaging software used to
render the data sets 3-dimensionally. This technique of studying
living plants is especially powerful when used in conjunction with 3D
representations of apicies that have been produced from fixed and
cleared samples; very accurate meristem maps have been produced.
Enhancer trapping is employed in gene discovery by mapping and
sequencing the t-DNA insertion region within the plant genome. Gene
expression patterns are thereby visualized and the identity and
function of the gene that the enhancer element would normally promote
can be discerned. Finally, enhancer trap plants can be secondarily
transformed so that any protein of interest can be expressed in the
same tissue-specific manner as the fluorescent protein. This opens up
many avenues of research including tissue perturbation or ablation by
misexpression of normally unexpressed, foreign, or lethal proteins.
Key words: Arabidopsis thaliana, development, enhancer trap, fluorescent protein, GFP, meristem