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Cotransformation and differential expression of introduced genes into potato (Solanum tuberosum L.) cv Bintje
Authors:M P Ottaviani  Ch H Hänisch ten Cate
Institution:(1) Center for Plant Breeding and Reproduction Research (CPRO), PO Box 16, NL-6700 AA Wageningen, The Netherlands
Abstract:Summary The Dutch potato cultivar Bintje has been transformed by Agrobacterium strain LBA1060KG, which contains two plasmids carrying three different DNAs (TL- and TR-DNA on the Agrobacterium rhizogenes plasmid and TKG-DNA on the pBI121 plasmid). Several transformed root clones were obtained after transformation of leaf, stem, and tuber segments, and plants were then regenerated from these root clones. The expression of the various marker genes rol, opine, beta-glucuronidase (GUS), and neomycin phosphotransferase (NPTII)] was determined in several root clones and in regenerated plants. The selection of vigorously growing root clones was as efficient as selection for kanamycin resistance. In spite of the location of NPTII and GUS genes on the same T-DNA, 17% of the root clones did not show GUS activity. Nevertheless, Southern blot analysis showed that these root clones contained at least three copies of the GUS gene. Sixty-four per cent of the root clones contained opines. The expression of these genes, however, was negatively correlated with plant regeneration capacity and normal plant development. The differential expression of the marker genes in the transgenic potato tissues is discussed.
Keywords:Agrobacterium rhizogenes  Binary vector  Cotransformation  GUS gene  Kanamycin resistance gene
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