ISSN:
1432-203X
Keywords:
Keywords Zygote
;
In vitro culture
;
Electroporation
;
Transformation
;
Tobacco
;
AbbreviationsNAA: Naphthaleneacetic acid
;
¶BA: 6-Benzyladenine
;
GUS: β-Glucuronidase
;
¶GFP: Green fluorescent protein
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
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.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/s002990000249
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