caa a22 4500 475795032 CHVBK 20180406123721.0 cr unu---uuuuu 170329e20001201xx s 000 0 eng 10.1007/s001220051604 doi (NATIONALLICENCE)springer-10.1007/s001220051604 Characterization and molecular analysis of transgenic plants obtained by microprotoplast fusion in sunflower [Elektronische Daten] [P. C. Binsfeld, R. Wingender, H. Schnabl] Abstract : Asymmetric somatic hybrid (ASH) plants were obtained by PEG-mediated mass fusion of microprotoplasts from perennial Helianthus species and hypocotyl protoplasts of Helianthus annuus. The formation of micronuclei in perennial sunflower cell cultures was induced, at early log phase, by addition of the herbicides amiprophos-methyl or oryzalin. Sub-diploid microprotoplasts were isolated by high-speed centrifugation and the smallest enriched by sequential filtration through nylon sieves of decreasing pore size. Fusion products were cultured and the regenerated plants phenotypically, genetically and cytologically characterized. DNA analysis using RAPD markers revealed that 28 out of 53 regenerated plants were asymmetric hybrids. Subsequent nuclear-DNA flow cytometric analysis showed that these plants had a higher DNA content than the receptor H. annuus, suggesting that they represented addition lines. Cytological investigation of the metaphase cells of 16 hybrids revealed an addition of 2-8 extra chromosomes in these plants. The phenotype of most ASH plants resembled H. annuus. These results indicate that micronuclear induction and asymmetric somatic hybridization represent a potent tool for partial genome transfer aimed at the specific transfer of economically important traits in breeding programs. Springer-Verlag Berlin Heidelberg, 2000 Key words Micronuclei nationallicence Microprotoplasts nationallicence Chromosome transfer nationallicence RAPD nationallicence Helianthus nationallicence Binsfeld P. C. Center for Biotechnology, Federal University of Pelotas, C.P 354, CEP 96010-900, Brazil, Present address: Institute of Agricultural Botany, Department of Physiology and Biotechnology of Plants, University of Bonn, Karlrobert-Kreiten-Strasse 13, D-53115 Bonn, Germany, BR aut Wingender R. Institute of Agricultural Botany, Department of Physiology and Biotechnology of Plants, University of Bonn, Karlrobert-Kreiten-Strasse 13, D-53115 Bonn, Germany, DE aut Schnabl H. Institute of Agricultural Botany, Department of Physiology and Biotechnology of Plants, University of Bonn, Karlrobert-Kreiten-Strasse 13, D-53115 Bonn, Germany, DE aut https://doi.org/10.1007/s001220051604 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s001220051604 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Binsfeld P. C. Center for Biotechnology, Federal University of Pelotas, C.P 354, CEP 96010-900, Brazil, Present address: Institute of Agricultural Botany, Department of Physiology and Biotechnology of Plants, University of Bonn, Karlrobert-Kreiten-Strasse 13, D-53115 Bonn, Germany, BR aut NATIONALLICENCE 700 1- Wingender R. Institute of Agricultural Botany, Department of Physiology and Biotechnology of Plants, University of Bonn, Karlrobert-Kreiten-Strasse 13, D-53115 Bonn, Germany, DE aut NATIONALLICENCE 700 1- Schnabl H. Institute of Agricultural Botany, Department of Physiology and Biotechnology of Plants, University of Bonn, Karlrobert-Kreiten-Strasse 13, D-53115 Bonn, Germany, DE aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer