Selection of transgenic flax plants is facilitated by spectinomycin |
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Authors: | Bérénice Bretagne-Sagnard Yves Chupeau |
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Institution: | (1) Laboratoire de Biologie Cellulaire, INRA, Route de Saint-Cyr, 78026 Versailles Cedex, France |
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Abstract: | Regeneration of transformed flax shoots after inoculation withAgrobacterium tumefaciens carrying a binary vector with either a neomycin phosphotransferase (nptII) gene and a -glucuronidase (GUS) reporter gene or a spectinomycin resistance gene was examined. Hypocotyls from 4-day-old seedlings were inoculated with either of the twoA. tumefaciens strains. Selection and regeneration were achieved on a medium containing 0.1 M thidiazuron, 0.01 M napthalene acetic acid, 100 mgl–1 kanamycin sulphate or spectinomycin sulphate and 300 mgl–1 cefotaxime. Use of different neomycins for the selection of transformed tissues did select transformed calli but not transformed shoots either directly or via a callus phase. Selection based on spectinomycin resistance allowed the growth of transformed shoots. Transgenic shoots were rooted on a medium containing 100 mgl–1 spectinomycin sulphate. Integration of the spectinomycin resistance gene into the flax genome was confirmed by Southern blot hybridizations and spectinomycin resistance was shown to be inherited as a dominant Mendeliant trait. Therefore, spectinomycin resistance is more suitable for genetic engineering of flax than aminoglycoside resistance. |
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Keywords: | Agrobacterium tumefaciens hypocotyl gene transfer flax neomycin spectinomycin |
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