Abstract
We have established a technique for isolating, culturing and transforming tobacco zygotes. Zygotes were isolated by microdissection or enzymatic maceration from fertilized embryo sacs. Viable zygotes cocultured with mesophyll protoplasts underwent first division after 3 days of culture. Zygotes isolated by microdissection underwent a higher frequency of first division (61.2%) than those isolated by enzymatic maceration (30.5%). Globular embryos were formed only from microdissected zygotes, at a frequency of 8.7% after 1–2 weeks in culture. An efficient millicell device for the electroporation of DNA into zygotes was established. The electroporated zygotes divided in vitro at a frequency of 54.6% and developed into proembryos. Introduced GFP gene constructs showed transient expression in about 2.6% of the electroporated tobacco zygotes.
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Received: 2 February 2000 / Revision received: 6 April 2000 / Accepted: 24 May 2000
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Li, S., Yang, H. Gene transfer into isolated and cultured tobacco zygotes by a specially designed device for electroporation. Plant Cell Reports 19, 1184–1187 (2000). https://doi.org/10.1007/s002990000249
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DOI: https://doi.org/10.1007/s002990000249