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Direct gene transfer to plant protoplasts by mild sonication

Summary

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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Abbreviations

CAT:

chloramphenicol acetyltransferase

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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

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Keywords

  • Chloramphenicol
  • Sugar Beet
  • Direct Gene
  • Transient Expression
  • Sucrose Concentration