Model system for plant cell biology: GFP imaging in living onion epidermal cells.
The ability to visualize organelle localization and dynamics is very useful in studying cellular physiological events. Until recently, this has been accomplished using a variety of staining methods. However, staining can give inaccurate information due to nonspecific staining, diffusion of the stain or through toxic effects. The ability to target green fluorescent protein (GFP) to various organelles allows for specific labeling of organelles in vivo. The disadvantages of GFP thus far have been the time and money involved in developing stable transformants or maintaining cell cultures for transient expression. In this paper, we present a rapid transient expression system using onion epidermal peels. We have localized GFP to various cellular compartments (including the cell wall) to illustrate the utility of this method and to visualize dynamics of these compartments. The onion epidermis has large, living, transparent cells in a monolayer, making them ideal for visualizing GFP. This method is easy and inexpensive, and it allows for testing of new GFP fusion proteins in a living tissue to determine deleterious effects and the ability to express before stable transformants are attempted.
Scott
A
A
North Carolina State University, Raleigh, NC, USA.
Wyatt
S
S
Tsou
P L
PL
Robertson
D
D
Allen
N S
NS
D003001:Cloning, Molecular; D018506:Gene Expression Regulation, Plant; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D008954:Models, Biological; D019697:Onions / Q000166:cytology* / Q000235:genetics; D015388:Organelles / Q000502:physiology; D010940:Plant Proteins / Q000235:genetics; D013194:Staining and Labeling / Q000379:methods; D014170:Transformation, Genetic
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D010940:Plant Proteins; D049452:Green Fluorescent Proteins