Oxidoreduction sites and relationships of spiroplasmas with insect cells in culture.

We have previously shown that Spiroplasma citri oxidoreduction sites, as revealed by the reduction of potassium tellurite into electron-dense tellurium crystals detectable by electron microscopy, were located at the blunt end of the organisms. The time of incubation of S. citri in potassium tellurite had no influence on the labeling. We have investigated the presence and location of oxidoreduction sites for other spiroplasmas such as the corn stunt spiroplasma (CSS), 277F, B88, BNR1 and PPS1. For all of these strains (except CSS, which is similar to S. citri), the location of oxidoreduction sites was affected by the time of incubation in potassium tellurite, and could be observed in different locations of the helix. 277F showed first labeling at the blunt end and a second, strong labeling at the tapered end. The other strains showed labeling all along the helix, but labeling at the blunt end generally appeared first. When Drosophila (Dm-1) cell cultures were infected with S. citri or 277F, the organisms adsorbed to the cells. Observation of infected cells by electron microscopy revealed that S. citri attached to the cells by the blunt end, while 277F attached either by the blunt end or the tapered one. The infection of leafhopper cell cultures (AS-2) with S. citri has been followed by transmission electron microscopy after incorporation of tritiated thymidine in the organism, and an immunocytochemical method has been developed to locate the organisms inside the cells.