Abstract
The present study reports a procedure for efficient microinjection of tobacco mosaic virus (TMV) into tomato callus cells. The ‘plate-culture system’ was devised for injection and incubation of the cells. In the system, the cells were embedded in the solid medium and cytoplasmic streaming (CS) of the cells was directly observed with the aid of a phase contrast microscope of the injectoscope. The cells which showed the active CS were selected for injection. Success of injection was judged by a maintenance of CS in the cells after the glass pipette inserted was removed. Multiplication of TMV in the injected cells was detected by staining with fluorescein isothiocyanate-conjugated antibody. When the cells were stained soon after TMV injection, fluorescent cells were not detected. However, TMV-injected cells were stained when incubated for 2 days after injection. These results suggest that the present method is suitable for inoculating tomato callus cells with TMV and obtaining a high level of TMV multiplication in injected cells.
