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  • 1
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell reports 19 (2000), S. 1041-1048 
    ISSN: 1432-203X
    Schlagwort(e): Key words Cassava (Manihot esculenta Crantz) ; Transformation ; Biolistics ; Positive selection ; Negative selection ; Abbreviations2,4-D 2,4-Dichlorophenoxyacetic acid ; BA 6-Benzylaminopurine ; CBM Basic medium ; CEM Elongation medium ; CIM Embryo induction medium ; CMM Maturation medium ; COM Organogenesis medium ; CTM Transformation medium ; GD Gresshoff and Doy medium ; IBA Indole-3-butyric acid ; FEC Friable embryogenic callus ; NAAα-Naphthalene acetic acid ; MSM Solid mannose selection medium ; MSN FEC Formation and embryo conversion medium ; SCV Settled cell volume ; SH Schenk and Hildebrandt medium ; SHM Liquid mannose selection medium
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  In order to develop new selection systems for production of transgenic cassava (Manihot esculenta Crantz), two different selection regimes were assessed for their efficiency on regeneration of transgenic cassava plants: positive selection using mannose and negative selection using hygromycin. Explants from somatic cotyledons and embryogenic suspensions were used as target tissues in the transformation experiments and bombarded using the particle inflow gun. Different culture and selection strategies were assessed to optimise the selection protocols. For the first time transgenic plants could be obtained using positive, and in the case of embryogenic suspensions, hygromycin-based negative selection. The stably transformed nature of the regenerated cassava plant lines and the expression of the transgenes were verified with PCR, RT-PCR, Southern and northern analyses. A rooting test for transgenic plants on a medium supplemented with mannose was developed to further improve the efficacy of the positive selection system. Our results demonstrate that it is possible to obtain transgenic cassava plants using non-antibiotic positive selection.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell reports 19 (2000), S. 939-945 
    ISSN: 1432-203X
    Schlagwort(e): Key words Cassava ; (Manihot esculenta Crantz) ; Transformation ; Biolistics ; Organogenesis ; Rooting test
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  A novel protocol, based on biolistics and regeneration via organogenesis, was developed for genetic transformation of cassava (Manihot esculenta Crantz). The in vitro performance of cassava cultivars CMC40, MPer183 and MCol22 was evaluated, and the regeneration protocol was modified to improve shoot production from explants for transformation experiments. Somatic cotyledons were used as a target tissue in the transformation experiments using the Particle Inflow Gun and a plasmid containing the uidA gene in transient assays. The effect of different parameters for particle bombardment efficiency, including the amount of DNA used, the flying distance of the projectiles and the pre- and post-plasmolysis time of the target tissue, was evaluated and the conditions were partially optimised. Stably transformed cassava plants of cvs. MCol22 and TMS60444 were produced using the partially optimised conditions and two different vector constructs carrying the hpt gene as the selectable marker. The selection protocol was optimised further, and a rooting test was developed for screening the regenerants for antibiotic resistance to reduce the number of escapes obtained after primary selection. The production of stably transformed cassava lines and the expression of the transgenes was verified by Southern blot analysis and RT-PCR.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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