A gfp reporter plasmid to visualize Azorhizobium caulinodans during nodulation of Sesbania rostrata.
Compared with other labeling techniques, the use of the green fluorescent protein (GFP) is advantageous to visualize bacteria because observations can be performed in real time. This feature is particularly interesting to study invasion events of rhizobia during nodule development on their legume host plant. To investigate the symbiotic interaction between Azorhizobium caulinodans ORS571 and Sesbania rostrata, we constructed two plasmids, pMP220-hem-gfp5 and pBBR5-hem-gfp5-S65T, that carry a modified gfp gene, the expression of which is controlled by the constitutive hem promoter. Introduction of either of these plasmids into A. caulinodans allowed the visualization of single bacteria. Determination of the plasmid stability in cultured bacteria and in nodules demonstrated that pBBR5-hem-gfp5-S65T is more stable than pMP220-hem-gfp5. The plasmid pBBR5-hem-gfp5-S65T can be used to study early invasion events during nodule development on hydroponic roots of S. rostrata.
D'Haeze
Wim
W
Department of Plant Systems Biology, Flanders Interuniversity Institute for Biotechnology (VIB), Ghent University, Technologiepark 927, B-9052, Belgium.
Gao
Mengsheng
M
Holsters
Marcelle
M
D020599:Azorhizobium caulinodans / Q000502:physiology*; D017931:DNA Primers; D007887:Fabaceae / Q000382:microbiology*; D017930:Genes, Reporter / Q000235:genetics*; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D015336:Molecular Probe Techniques; D018517:Plant Roots / Q000382:microbiology*; D010957:Plasmids / Q000235:genetics*
D017931:DNA Primers; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins