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Regeneration of transgenic cassava plants (Manihot esculenta Crantz) through Agrobacterium-mediated transformation of embryogenic suspension cultures
Authors:A E González  C Schöpke  N J Taylor  R N Beachy  C M Fauquet
Institution:(1) International Laboratory for Tropical Agricultural Biotechnology (ILTAB/ORSTOM-TSRI), Division of Plant Biology – BCC 206, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA Fax: +01-619-784-2994 e-mail: ILTAB@Scripps.Edu, US
Abstract:A protocol was developed for Agrobacterium-mediated transformation of embryogenic suspension cultures of cassava. The bacterial strain ABI containing the binary vector pMON977 with the nptII gene as selectable marker and an intron-interrupted uidA gene (encoding β-glucuronidase) as visible marker was used for the experiments. Selection of transformed tissue with paromomycin resulted in the establishment of antibiotic-resistant, β-glucuronidase-expressing lines of friable embryogenic callus, from which embryos and subsequently plants were regenerated. Southern blot analysis demonstrated stable integration of the uidA gene into the cassava genome in five lines of transformed embryogenic suspension cultures and in two plant lines.
Keywords:Embryogenic suspension cultures  β  -Glucuronidase  Agrobacterium tumefaciens  Cassava  Paromomycin
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