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Efficient production of transgenic plants by Agrobacterium-mediated transformation of cassava (Manihot esculenta Crantz)
Authors:MM Schreuder  CJJM Raemakers  E Jacobsen  RGF Visser
Institution:(1) The Graduate School of Experimental Plant Sciences, Laboratory of Plant Breeding, Wageningen University and Research Center, P.O. Box 386, 6700 AJ Wageningen, The Netherlands
Abstract:An efficient and reproducible method was developed for Agrobacterium-mediated transformation of embryogenic suspension cultures of cassava. LBA4404(pTOK233), containing the nptII, hph and gus marker genes, was used in the experiments. Chemical selection by means of kanamycin was used to establish 1037antibiotic resistant callus lines, of which 526 showed GUS expression. Of the 241 callus lines that were transferred to maturation medium 219formed somatic embryos. Thirty-seven of the 38 lines that were transferred to germination medium produced plants. GUS-positive plants could be obtained from 31 lines; in 14 of those lines 100% of the produced plants were GUS-positive, the remaining 17 lines yielded GUS-positive plants at an average of 72%. The transgenic nature of these plants was confirmed by Southern blot analysis. This revised version was published online in July 2006 with corrections to the Cover Date.
Keywords:Agrobacterium tumefaciens            cassava  friable embryogenic callus  genetic modification  hygromycin  kanamycin
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