Stable genetic transformation of mature wheat embryos using silicone carbide fibers and DNA imbibition

• Autorzy: Sawahel W. , Saker M.
• Języki publikacji: EN

Abstrakty

EN

Improvement of wheat (Triticum aestivum L) by biotechnological approaches is currently limited by a lack of tissue culture - free transformation system. In this paper, we present the development of a gene transfer system using DNA imbibition [DI]; and/or silicone carbide fibers [SCF] treatments for gene delivery and seed-derived embryos as gene target. Plasmid DNA (pBI221.23), containing the selectable "hpt" gene for hygromycin resistance and the reporter "gus" gene, was delivered into mature wheat embryos via [DI] and/or [SCF] treatments. In a histochemical analysis of β-GUS activity, an average of 13.8 % of the mature embryos receiving the combined treatment [DI + SCF] showed gus expression, whereas only 3.2% and 0.4% showed gus expression following [SCF] and [DI] respectively. The mature embryos also showed more multi-site gus expression (4.6%) than those treated only with [SCF] (0.4%). The growth of the produced plantlets on a medium containing 45 µg/ml hygromycin B indicated that the transformation efficiency of the combined treatments [DI + SCF] was 0.8% in comparison to 0.1% for that of [SCF] treatment and no hygromycin resistant-plantlets were detected following [DI]. Stable transformation was confirmed through polymerase chain reaction "PCR" and Southern hybridization which indicated that a functional hpt gene had integrated into wheat chromosomal DNA.

Słowa kluczowe

EN

DNA imbibition; transient expression; silicone carbide fibre; Triticum aestivum; polymerase chain reaction; wheat embryo; stable transformation; gene transfer system; plasmid DNA

• Wydawca: -
• Czasopismo: Cellular and Molecular Biology Letters
• Rocznik: 1997
• Tom: 02
• Numer: 4
• Opis fizyczny: p.421-429,fig.

Twórcy

autor

Sawahel W.

• National Research Center, Cairo, Egypt

autor

Saker M.

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