Organelle Identification and Characterization in Plant Cells: Using a Combinational Approach of Confocal Immunofluorescence and Electron Microscope

• Published in: Journal of plant biology = Singmul Hakhoe chi Vol. 52; no. 1; pp. 1 - 9
• Main Authors: Tse, Yu Chung, The University of Chicago, Chicago, IL, USA, Lam, Sheung Kwan, University of California, Berkeley, Berkeley, CA, USA, Jiang, Liwen, The Chinese University of Hong Kong, Hong Kong, China
• Format: Journal Article
• Language: English
• Published: New York Springer-Verlag 01.02.2009; Springer Nature B.V; 한국식물학회
• Subjects: Biomedical and Life Sciences; Cells (biology); Compartments; Confocal microscopy; CYTOPLASMIC ORGANELLES; Dynamic response; electron microscope; Electron microscopes; Endoplasmic reticulum; Endosomes; Fluorescence; Golgi apparatus; Identification; Immunofluorescence; Kinases; Life Sciences; Organelles; ORGANITE CELLULAIRE; ORGANULOS CITOPLASMICOS; Plant Breeding/Biotechnology; Plant cells; Plant Ecology; Plant Genetics and Genomics; Plant Sciences; Plant Systematics/Taxonomy/Biogeography; Proteins; Review Article; Tobacco; Transgenic plants; Vacuoles; Yeast; 생물학; Golgi Apparatus; Immunogold Electron Microscope; Internal Vesicle; Green Fluorescent Protein Fusion; Wortmannin
• ISSN: 1226-9239; 1867-0725
• DOI: 10.1007/s12374-008-9000-z

The plant secretory and endocytic pathways consist of several functionally distinct membrane-bounded compartments. The ultra structures of the endoplasmic reticulum, the Golgi apparatus, and central vacuoles have been well characterized via traditional structural electron microscope (EM). However, the identification of plant prevacuolar compartments (PVCs) and early endosomes (EEs) had not been achieved until more recently because of the lack of specific markers for these organelles. Recent development of fluorescent reporters for PVCs and EEs expressing in transgenic tobacco BY-2 cells and Arabidopsis plants has allowed their dynamic characterization in living cells via confocal microscopy and drug treatment, which led to their subsequent morphological identification via structural and immunogold EM. Thus, in this review, we will use our studies on PVCs and EEs as examples to present an efficient approach for organelle identification in plant cells via primary characterization of fluorescent-marked organelles in living cells and their dynamic response to drug treatments, which then serves as the basis for subsequent immunogold and structural EM studies for organelle identification. Such strategy thus represents a powerful approach in future research for the identification of novel organelles and transport vesicles in plant cells.