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  • 1
    Electronic Resource
    Electronic Resource
    Factors promoting sustained divisions of mesophyll protoplasts isolated from dihaploid clones of potato (Solanum tuberosum L.) and a cytological analysis of regenerated plants (1991)
    Springer
    Plant cell, tissue and organ culture 27 (1991), S. 257-265 
    Details
    ISSN: 1573-5044
    Keywords: cytological analysis ; dihaploid potato ; protoplast culture ; shoot regeneration ; sustained division
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A culture protocol has been developed for mesophyll protoplasts isolated from various dihaploid clones of potato. A special effort was made to promote the growth of initially dividing cells to form cell colonies and calli. An increase in plating efficiency in 3 different dihaploid clones and one doubled dihaploid clone was obtained after serial dilution of cultures with a suitable amount and type of medium at different stages of cell colony development. Plating on a refined semi-solid medium after 14 days of culture further improved both the yield and the quality of calli obtained. The refined plating medium also enhanced shoot regeneration ability from 67 to 90% in one of the dihaploid clones (67:9). The refined culture protocol could also be used without causing a decrease in plating efficiency at a low population density adjusted after 3 days of culture. The ploidy level of plants regenerated from dihaploid protoplasts were determined by chromosome counting and DNA analysis by flow cytometry. Most of the plants were aneuploid or tetraploid although, some dihaploid plants were obtained after protoplast culture of 2 dihaploid clones derived from the same cultivar (cv. Stina).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00157589
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Survival and growth of pea protoplasts after transformation by electroporation (1992)
    Springer
    Plant cell, tissue and organ culture 30 (1992), S. 141-148 
    Details
    ISSN: 1573-5044
    Keywords: electroporation ; pea ; Pisum sativum L. ; protoplast ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts from two different pea cultivars, Belman and Filby, were stably transformed by direct gene transfer using electroporation. Transgenic calli could be obtained after selection, when hygromycin resistance was used as the selective trait introduced into the protoplasts, while no transformants were obtained when kanamycin resistance was used as selective marker in either of the two pea cultivars tested. The effect of the field strength on survival and division rates of the protoplasts was studied. Two different culture systems and osmotica were compared for induction of sustained divisions in and regeneration of transgenic callus from the protoplasts. The choice of the culture system had a considerable effect on the initial division frequency of the treated protoplasts, as well as on the later growth of the colonies. Transformation efficiency was monitored by histochemical GUS assay, and the transgenic nature of the calli selected for resistance against antibiotics was confirmed by DNA analysis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00034308
    Library Location Call Number Volume/Issue/Year Availability
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    Paper (German National Licenses)
    Fulltext
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