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  • 1
    Electronic Resource
    Electronic Resource
    Protoplasts from cotyledon and hypocotyl of sunflower (Helianthus annuus L.): shoot regeneration and seed production (1992)
    Springer
    Plant cell reports 11 (1992), S. 632-636 
    Details
    ISSN: 1432-203X
    Keywords: Helianthus annuus ; Protoplasts ; Plant regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cotyledon and hypocotyl protoplasts of Helianthus annuus inbred line 47 302 bcd were embedded in alginate and plated on L4 medium (Lenée and Chupeau 1986). After one month, the calli were transferred on MSSH regeneration medium (Murashige and Skoog 1962; Schenk and Hildebrandt 1972) where they regenerated shoots (overall efficiency 10−2%). The shoots were elongated on B5 (Gamborg et al. 1968) medium first without hormones, then supplemented with GA3 and BAP (both 0.05 mg/l). In order to overcome the difficulty to induce rooting by classical methods, the elongated shoots were grafted on a sunflower rootstock. The grafted shoots produced flowers and seeds. Different factors have been shown to have an important influence on the capacity to regenerate shoots: the genotype, the physical culture conditions at the callus regeneration step (e.g. protoplasts embedded in alginate), and the media composition.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00236388
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Visualisation of transgene expression at the single protoplast level (1992)
    Springer
    Plant cell reports 11 (1992), S. 346-350 
    Details
    ISSN: 1432-203X
    Keywords: Beet Necrotic Yellow Vein Virus ; Chenopodium quinoa ; Coat protein ; Frequency of expression ; Nicotiana tabacum ; NPT II ; Protoplast printing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts are currently used to study the expression of genes following transformation. Expression is followed on a population of protoplasts after total protein extraction by conventional western blotting or measure of the enzymatic activity of the transgenic protein. We describe here a new method, called protoplast printing, allowing easy detection of the fraction of cells expressing a certain protein within a population of protoplasts. It consists of immobilization of the protoplast proteins on a nitrocellulose filter, so as to retain the outlines of the cell, followed by immunological detection of the protein of interest. The only special requirement is an antibody specific for the protein. We have studied the expression of the BNYVV coat protein after electroporation of Chenopodium quinoa protoplasts with viral RNAs, and the expression of the NPT II gene in protoplasts isolated from transgenic tobacco plants as well as after direct transfer of plasmid DNA into tobacco protoplasts. In both cases — infection with viral RNAs and transformation with plasmid DNA — expressing and non-expressing cells can be distinguished as early as 12h after transfer of the transgenes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233363
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    Paper (German National Licenses)
    Fulltext
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