ISSN:
1432-203X
Keywords:
Datura
;
haploids
;
direct gene transfer
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract We developed a highly efficient transformation protocol for the PEG-mediated direct transfer of plasmid DNA into protoplasts of haploid Datura innoxia. Vectors harbouring a neomycin phosphotransferase II gene or a hygromycin B phosphotransferase gene under the control of different promoters were used in the transformation experiments. Various amounts of plasmid DNA were applied without any carrier DNA to show the direct influence of the plasmid DNA concentration on the transformation efficiency. Approximately 95% of the selected calli were regenerated to plants; 20% of them remained haploid. Total DNA of different transgenic plants was analysed with regard to the integration pattern of the plasmid DNA. Plants carrying only one or two copies of the vector DNA were observed as well as individuals with multi-copy integration (up to ten or more copies).
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00234698
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