Transgenic turf-type tall fescue (Festuca amndinacea Schreb.) plants regenerated from protoplasts |
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Authors: | Sam-Bong Ha Fang-Sheng Wu Tracy K. Thorne |
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Affiliation: | (1) Department of Crop and Soil Environmental Sciences, Virginia Polytechnic Institute and State University, 24061 Blacksburg, VA, USA;(2) Department of Biology, Virginia Commonwealth University, 23284 Richmond, VA, USA |
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Abstract: | To improve turfgrasses using genetic engineering, we have developed a transformation system in turf-type tall fescue, one of the most important turfgrass species. Embryogenic cell cultures were established after callus induction from embryos of mature seed. The agarose-bead method with nurse cells was used to culture protoplasts and plants were regenerated from protoplasts of tall fescue cultured cells. To develop transgenic tall fescue plants, the hygromycin resistance gene and the -glucuronidase gene were introduced into the tall fescue protoplasts by electroporation. A high concentration (200 mg/l) of hygromycin was required to select transformed cells because of the high level of endogenous resistance to the antibiotic in tall fescue. Most of the transformed cells exhibited GUS activity and several plants were regenerated from these cells. The presence of introduced genes was confirmed by Southern blot hybridization of PCR amplified DNA from transgenic plants.Abbreviations Adh alcohol dehydrogenase - BAP benzylaminopurine - bp base pair(s) - GUS -glucuronidase - Kb kilobase(s) - MS Murashige and Skoog's medium - PCR polymerase chain reaction |
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