Abstract
Direct gene transfer into plant protoplasts has been recently developed, and conditions for high frequency transformation of SR1 tobacco protoplasts established. In this paper we analyse numerous transformation parameters in a comparative study on SR1 Nicotiana tabacum and N. plumbaginifolia, and report on a simple chemical technique for very efficient protoplast transformation. It is based on the synergistic interaction of MgCl2 and PEG. The technique yielded up to 1400 transformants per 3×105 treated N. tabacum protoplasts (up to 4.8% of the survivors, late selected clones). Using N. plumbaginifolia, the frequencies were 10-fold lower, indicating that the ‘competence’ for transformation has a species-specific component.
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Abbreviations
- PEG:
-
polyethyleneglycol
- RTF:
-
relative transformation frequency
- ATF:
-
absolute transformation frequency
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Negrutiu, I., Shillito, R., Potrykus, I. et al. Hybrid genes in the analysis of transformation conditions. Plant Mol Biol 8, 363–373 (1987). https://doi.org/10.1007/BF00015814
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DOI: https://doi.org/10.1007/BF00015814