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High Efficiency Electro-transfection of Tobacco Mesophyll Protoplasts with Tobacco Mosaic Virus RNA

National Institute of Agrobiological Resources, Tsukuba Science City, Yatabe, Ibaraki 305
and¹ Japan Spectroscopic Co. Ltd, Ishikawa 2967-5, Hachioji, Tokyo 192, Japan

Tobacco mesophyll protoplasts were transfected with tobacco mosaic virus (TMV) RNA in an electric field using a newly devised chamber constructed with gold-coated glass panel electrodes. Up to 95% of protoplasts subjected to several DC pulses (50 µs, 550 to 800 V/cm) while suspended in 0.5 M-mannitol containing 10 µg/ml TMV RNA became infected. The treatment did not affect the viability of the protoplasts or their stability during 40 h of incubation.

Keywords: TMV RNA, electro-transfection, transfection chamber, tobacco protoplast

Received 5 February 1986; accepted 24 June 1986.