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  • 1
    ISSN: 1432-2242
    Schlagwort(e): Microspore protoplasts ; Somatic protoplasts ; Fusions ; Triploids ; Nicotiana
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Microspore protoplasts (n) isolated at the tetrad stage from plants of Nicotiana tabacum Km+ (2n=4x=48) were fused with somatic cell protoplasts (2n) of WT N. rustica (2n=4x=48) to produce triploid plants. A total of 21.2×106 microspore protoplasts were fused with 11.2×106 somatic cell protoplasts using the high pH/Ca+ + method. Microspore protoplasts did not divide and WT N. rustica protoplasts stopped dividing when the protoplast-derived colonies were transferred to a selection medium containing kanamycin. A total of 104 actively growing green colonies were recovered on the selection medium. Ninety-six of these colonies were tested for their hybrid nature by PAGE of peroxidases and were found to contain bands characteristic of both parents. Hybrid nature of the plants regenerated from some of the selected colonies was confirmed by IEF of leaf esterases, by NPT II activity assay and by hybridizing total DNAs restricted with EcoR I to a cloned 18s rDNA fragment. Root tip squashes of six of the hybrid plants revealed chromosome numbers ranging from 58–72. From chromosomal and biochemical analysis, it can be concluded that the procedure of fusing microspore protoplasts (n) of species A carrying a dominant selection marker with WT somatic cell protoplasts (2n) of species B can be a convenient selection method for the synthesis of triploid plants. The significance of triploids lies in their subsequent use for transferring alien chromosomes and genes of species A to species B.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 78 (1989), S. 547-552 
    ISSN: 1432-2242
    Schlagwort(e): Microspore protoplasts ; Protoplast fusion ; Gametosomatic hybrids ; Organelle transmission ; Nicotiana species
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Gametosomatic hybrids produced by the fusion of microspore protoplasts of Nicotiana tabacum Km+Sr+ with somatic cell protoplasts of N. rustica were analysed for their organelle composition. For the analysis of mitochondrial (mt)DNA, species-specific patterns were generated by Southern hybridization of restriction endonuclease digests of total DNA and mtDNA with four DNA probes of mitochondrial origin: cytochrome oxidase subunit I, cytochrome oxidase subunit II, 26s rDNA and 5s-18s rDNA. Of the 22 hybrids analyzed, some had parental-type pattern for some probes and novel-type for the others, indicating interaction between mtDNA of the two parent species. For chloroplast (cp)DNA analysis, species-specific patterns were generated by Southern hybridization of restriction endonuclease digests of total DNA with large subunits of ribulose bisphosphate carboxylase and cpDNA as probes. All the hybrids had N. rustica-specific patterns. Hybrids were not resistant to streptomycin, a trait encoded by the chloroplast genome of N. tabacum. In gametosomatic fusions of the two Nicotiana species, mitochondria but not the chloroplasts are transmitted from the parent contributing microspore protoplasts.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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