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Production and identification of somatic hybrids between Solanum tuberosum and S. papita by using the rolC gene as a morphological selectable marker
Authors:C Kaendler  M Fladung  H Uhrig
Institution:(1) Max-Planck-Institut für Züchtungsforschung, Abteilung Pflanzenzüchtung und Ertragsphysiologie, D-50829 Köln, Germany;(2) Present address: Organic Waste Systems, Dok Noord 4, B-9000 Gent, Belgium;(3) Present address: Bundesforschungsanstalt für Forst und Holzwirtschaft, Institut für Forstgenetik, Sieker Landstr 2, D-22927 Großhansdorf, Germany
Abstract:A successful hybridization of a diploid clone of Solanum tuberosum with a rolC-transgenic, diploid S. papita clone is reported. By using leaf expiants of this S. papita clone, which after transformation expressed kanamycin resistance, intact protoplasts were obtained, but these protoplasts did not develop to microcalli or regenerate to mature plants. However, protoplasts of the S. tuberosum clone showed a high capacity to regenerate plants from isolated protoplasts. On a medium containing Kanamycin only calli regenerated to plants, which revealed a rolC phenotype (reduced apical dominance with a large number of adventitious shoots and a pale green color of leaves) and later on turned out to be true hybrids. Self fusions of S. papita never developed to microcalli and those of S. tuberosum ceased to develop on the kanamycin-containing medium. Identification of somatic hybrids was done by RFLP and RAPD analysis. In the greenhouse, out of four selected hybrids only FK3.1 was successfully crossed with two standard S. tuberosum varieties (Datura, Desirée). Out of all the seeds germinated, only rolC-negative F1 seedlings were further characterized. Within the seedling population obvious differences were evident in respect of the S. papita and S. tuberosum characteristics.
Keywords:Marker genes  Protoplast fusion  RolC  Somatic hybrid  Transgenic potato
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