|Monier, Jean-Michel - UNIV.CLAUDE-BERNARD, FR.|
Submitted to: CRC Press
Publication Type: Book / Chapter
Publication Acceptance Date: January 1, 2005
Publication Date: April 1, 2006
Citation: Brandl, M., Monier, J. 2006. Methods in microscopy for the visualization of microbes and their behavior on plants. CRC Press LLC. Ames Iowa.595-619 Interpretive Summary: The discovery of confocal microscopy, and the green fluorescent protein (GFP) as an intrinsic bacterial label, spurred a new revolution, starting in the 1990's, in the use of fluorescence microscopy to study bacteria in their natural habitat. Because most bacterial species or strains cannot be distinguished from each other microscopically, intrinsic labeling of bacteria with GFP or other fluorescent proteins has been used widely to track specific bacteria in complex environments, including plants. This chapter focuses on novel experimental approaches in fluorescence microscopy to detect bacteria and investigate their behavior on plants. Recent advances in microscope technologies that may be applied to plant microbiology research are discussed also.
Technical Abstract: The chapter focuses on methods in fluorescence microscopy that are useful for the investigation of the ecology of plant-associated, plant pathogenic, and human enteric bacteria on plants. More precisely, we discuss the various fluorescent proteins, dyes, and bioconjugates that are available to visualize the localization of bacteria on plants, as well as some of the advantages and pitfalls related to theses technologies. Applications of methods in fluorescence microscopy are presented also, such as the study of spatial distribution of bacterial cells on plants, of cell-cell interactions, and of bacterial gene expression in planta, and the measurement of biological parameters, e.g. cell viability and pH. Finally, recent advances in microscopy methods that may impact plant microbial ecology research is presented.