Detection of nitrogenase in individual cells of a natural population of trichodesmium using immunocytochemical methods for fluorescent cells

The intercellular distribution of nitrogenase was detected in a natural population of Trichodesmium spp. collected from Shitaba Bay, Uwajima, Ehime Prefecture, Japan. Polyclonal antibodies raised against the Fe-protein and the MoFe-protein (α-subunit) of nitrogenase were used as probes. Visualization of the antibody binding to the nitrogenase was performed using horseradish peroxidase-conjugated secondary antibody and the chromogenic substrate, 3,3′-diaminobenzidine tetrahydrochloride, to avoid the masking effects caused by the bright autofluorescence emitted by the Trichodesmium cells. Nitrogenase proteins were detected in 66 to 81% of the trichomes. More than 77% of the cells were immunostained in individual nitrogenase-containing trichome. The data show natural populations of Trichodesmium spp. do not developed heterocyst-equivalent cells.