Research

Multidimensional fluorescence microscopy of multiple organelles in Arabidopsis seedlings

Naohiro Kato¹ , Dexter Reynolds^1,2 , Matthew L Brown¹ , Marietta Boisdore¹ , Yukichi Fujikawa¹ , Andrea Morales³ and Lee A Meisel³

¹  Louisiana State University, Department of Biological Sciences, Baton Rouge, LA, 70803, USA

²  Southern University and A&M College, Baton Rouge, LA, 70813, USA

³  Millennium Nucleus in Plant Cell Biology and Center of Plant Biotechnology, Andres Bello University, Av. República 217, 837-0146 Santiago, Chile

author email corresponding author email

Plant Methods 2008, 4:9doi:10.1186/1746-4811-4-9

Published:	19 May 2008

Abstract

Background

The isolation of green fluorescent protein (GFP) and the development of spectral variants over the past decade have begun to reveal the dynamic nature of protein trafficking and organelle motility. In planta analyses of this dynamic process have typically been limited to only two organelles or proteins at a time in only a few cell types.

Results

We generated a transgenic Arabidopsis plant that contains four spectrally different fluorescent proteins. Nuclei, plastids, mitochondria and plasma membranes were genetically tagged with cyan, red, yellow and green fluorescent proteins, respectively. In addition, methods to track nuclei, mitochondria and chloroplasts and quantify the interaction between these organelles at a submicron resolution were developed. These analyzes revealed that N-ethylmaleimide disrupts nuclear-mitochondrial but not nuclear-plastids interactions in root epidermal cells of live Arabidopsis seedlings.

Conclusion

We developed a tool and associated methods for analyzing the complex dynamic of organelle-organelle interactions in real time in planta. Homozygous transgenic Arabidopsis (Kaleidocell) is available through Arabidopsis Biological Resource Center.