Gene transfer into isolated and cultured tobacco zygotes by a specially designed device for electroporation |
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Authors: | S T Li H Y Yang |
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Institution: | (1) Research Center for Developmental Biology, College of Life Science, Wuhan University, Wuhan 430072, China e-mail: hyyang@whu.edu.cn Fax: +86-27-87646010, CN |
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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.
Received: 2 February 2000 / Revision received: 6 April 2000 / Accepted: 24 May 2000 |
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Keywords: | Zygote In vitro culture Electroporation Transformation Tobacco |
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