A novel procedure employing mild sonication for transformation of plant protoplasts is described. Transient expression of a chloramphenicol acetyltransferase (CAT) gene in protoplasts of sugar beet (Beta vulgaris L.) and tobacco (Nicotiana tabacum L.) was obtained by a brief exposure of the protoplasts to 20 kHz ultrasound in the presence of plasmid DNA. Maximum levels of CAT activity were achieved by sonication for 500–900 ms at 30–70 W electric power (0.65–1.6 W/cm2 acoustic power). This reduced the viability to 15–20 % and 60 % for sugar beet and tobacco protoplasts, respectively. Up to 12 % (sugar beet) and 81 % (tobacco) of maximum transient expression could be achieved with no significant loss of viability. Protoplasts surviving exposure to ultrasound were found to have a similar long-term viability and to regenerate to micro-calli as untreated protoplasts. Plasmid DNA concentrations of 80–110 μg/ml and sucrose concentrations of 21–28 % in the sonication medium were found to be optimal for transient expression.
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Birnboim HC (1983) Meth. Enzymol. 100:243–255.
Chapman IV (1974) Br. J. Radiol. 47:411–415.
Ciaravino V, Miller MW, Kaufman GE (1981) Ultrasound Med. and Biol. 7:175–184.
Crossway A, Oakes JV, Irvine JM, Ward B, Knauf VC, Shewmaker CK (1986) Mol. Gen. Genet. 202:179–185.
Davey MR, Cocking EC, Freeman J, Pearce N, Tudor I (1980) Plant Sci. Lett. 18:307–313.
Fizzell LA (1988) Biological effects of acoustic cavitation. In: Suslick KS (ed) Ultrasound. Its chemical, physical and biological effects. VCH publishers, Weinheim, FRG. pp. 287–304.
Frearson EM, Power JB, Cocking EC (1973) Dev. Biol. 33:130–137.
Fromm ME, Taylor LP, Walbot V (1986) Nature 319:791–793.
Hain R, Stabel P, Czernilofsky AP, Steinbiss HH, Herrera-Estrella L, Schell J (1985) Mol. Gen. Genet. 199:161–168.
Joersbo M, Brunstedt J (1990a) Plant Cell Rep. In press.
Joersbo M, Brunstedt J (1990b) Ultrasound Med. and Biol. In press.
Klein TM, Wolf ED, Wu R, Sanford JC (1987) Nature 327:70–73.
Krens FA, Molendijk L, Wullens GJ, Schilperoort RA (1982) Nature 296:72–74.
Lindsey K, Jones MGK (1987) Plant Mol. Biol. 10:43–52.
Miller MW, Voorhees SM, Carstensen EL, Eames FA (1974) Radiat. Bot. 14:201–206.
Miller MW, Ciaravino V, Allen D, Jensen S (1976) Int. J. Radiol. Biol. 30:217–222.
Mortimer AJ, Dyson M (1988) Ultrasound Med. and Biol. 14:499–506.
Pietrzak M, Shillito RD, Hohn T, Potrykus I (1986) Nucl. Acids Res. 14:5857–5868.
Potrykus I, Paszkowski J, Saul MW, Petruska J, Shillito RD (1985) Mol. Gen. Genet. 199:169–177.
Shillito RD, Saul MW, Paszkowski J, Mueller M, Potrykus I (1985) Bio/Technol. 3:1099–1103.
Szabados L, Gaggero C (1985) Plant Cell Rep. 4:195–198.
Werr W, Loerz H (1986) Mol. Gen. Genet. 202:471–475.
Widholm JM (1972) Stain Technol. 47:189–194.
Zimmermann U, Pilwat G, Riemann F (1974) Biophys J. 14:881–899.
Zimmermann U, Pilwat G, Pequeux A, Gilles R (1980) J. Membrane Biol. 54:103–113.
Communicated by H. Lörz
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Joersbo, M., Brunstedt, J. Direct gene transfer to plant protoplasts by mild sonication. Plant Cell Reports 9, 207–210 (1990). https://doi.org/10.1007/BF00232181
- Sugar Beet
- Direct Gene
- Transient Expression
- Sucrose Concentration