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Plant and Cell Physiology, 1985, Vol. 26, No. 2 229-236
© 1985

Article

Capillary Microinjection into Protoplasts and Intranuclear Localization of Injected Materials

Hiromichi Morikawa and Yasuyuki Yamada

Research Center for Cell and Tissue Culture, Faculty of Agriculture, Kyoto University Kyoto 606, Japan

By a direct microinjection through pressurized glass micropipettes, we injected fluorescent Lucifer Yellow, berberine and berberine-bound DNA into protoplasts isolated from cultured Euphorbia millii cells 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.

(Received August 6, 1984; Accepted November 12, 1984)
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