Detection of chromosomes tagged with green fluorescent protein in live Arabidopsis thaliana plants

Naohiro Kato and Eric Lam

Biotech Center, Rutgers University, Cook College, 59 Dudley Rd, New Brunswick, NJ 08904, USA

author email corresponding author email

Genome Biology 2001, 2:research0045.1-0045.10doi:10.1186/gb-2001-2-11-research0045


Published:	12 October 2001

Subject areas: Molecular biology, Methods, Plant biology, Cell biology

Abstract

Background

Structural and dynamic studies of chromosomes tagged with green fluorescent protein (GFP) in yeast and cultured animal cells have revealed some surprises. Although this technology can be very powerful, only a few studies using this approach with developed multicellular systems have been reported for the study of chromatin behavior in situ.

Results

We established vectors and conditions to visualize tagged loci stably inserted in the Arabidopsis genome via GFP fused to a bacterial DNA-binding protein. Using this system, three-dimensional coordinates for tagged loci within nuclei from cells of a live plant can be directly determined with concomitant visualization of the position of the nucleolus. Chromosome polyploidization in epidermal cells at the elongation zone of the root in transgenic plants can be visualized in situ using this technique.

Conclusion

We have established that GFP fusion with DNA-binding proteins can be used in conjunction with concatameric binding-site arrays to track genomic loci in living Arabidopsis plants. It should now be feasible to study the mechanisms of organization and dynamics of chromatin in specific cell types during various times of plant development, taking advantage of the well developed genetic systems and resources available for Arabidopsis.