Capillary Microinjection into Protoplasts and Intranuclear Localization of Injected Materials

摘要

By a direct microinjection through pressurized glass micropipettes, we injected fluorescent Lucifer Yellow, berberine and berberine-bound DNA into protoplasts isolated from culturedEuphorbia milliicells and from tobacco mesophyll cells. The protoplasts to be injected were held by a holding pipette with which the cytoplasm or nucleus of the protoplasts could be oriented with reference to the tip of the injection pipette. Upon microinjection of a berberine-bound DNA, the nucleus and intranuclear organelles, possibly including nucleolus, specifically exhibited yellow fluorescence due to berberine. This provides a direct evidence that DNA molecules can be microinjected into the intranuclear compartment of the protoplasts. The injected protoplasts survived the capillary microinjection.