African Journal of Tropical Agriculture ISSN 2375-091X Vol. 7 (8), pp. 001-006, August, 2019. © International Scholars Journals
Full Length Research Paper
Detection of gfp expression from gfp-labelled bacteria spot inoculated onto sugarcane tissues
Joyce Prisca NJOLOMA1*, Moriya OOTA2, Yuichi SAEKI1 and Shoichiro AKAO1
1Department of Biochemistry and Applied Bioscience, Faculty of Agriculture, University of Miyazaki, Miyazaki, 889-2192 Japan.
2Okinawa Prefecture Agricultural Experimental station, Nishizato, Hirara. 906-0012, Japan.
Accepted 27 March, 2019
Green fluorescent protein (GFP) as a marker gene has facilitated biological research in plant-microbe interactions. However, there is one major limiting factor in the detection of GFP in living organisms whose cells emit background autofluorescence. In this study, Herbaspirillum sp. B501gfp1 bacterial cells were spot inoculated onto 5 month-old sterile micro-propagated sugarcane tissues to detect if the GFP fluorescence expression could be distinguished from the tissue’s background fluorescence. Stem tissues and leaf sections mounted on glass slides were directly inoculated with a single touch using the tip of a syringe previously dipped into the inoculum containing 108 bacterial cells/ml. We observed that GFP fluorescence could be easily distinguished in the stem than in the leaf tissues. However, the brightness level of the fluorescence varied with time as a result of fluctuations in the bacterial cell density. The presence of chloroplasts in the leaf tissues of sugarcane requires the use of bright GFP variants when monitoring bacteria-plant interactions using GFP labelled bacteria.
Key words: GFP fluorescence, autofluorescence, bacterial cell density, sugarcane plant tissue.