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  • 1
    Electronic Resource
    Electronic Resource
    Expression of foreign genes in sunflower (Helianthus annuus L.) — evaluation of three gene transfer methods (1955)
    Springer
    Euphytica 85 (1955), S. 63-74 
    Details
    ISSN: 1573-5060
    Keywords: Agrobacterium tumefaciens ; electroporation ; particle gun ; polyethylene glycol ; regeneration ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Suitable sunflower tissues and cells were transformed either by direct gene transfer into protoplasts, particle bombardment, or Agrobacterium co-culture. While all techniques allowed efficient short-term or transient expression of the introduced gene(s) in the respective tissues, stable transformation was only observed after transformation with Agrobacterium. The latter technique was suitable for the production of transgenic callus from seedling cotyledons and occasional shoots with chimaeric expression of the transgene. Detailed analysis of the interaction of Agrobacterium with this explant showed that infection efficiency was critically dependent on the co-culture conditions, and that the preferentially-transformed cells were not the ones competent for regeneration.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023931
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Gene specificity is maintained in transient expression assays with protoplasts derived from different tissues of barley (1955)
    Springer
    Euphytica 85 (1955), S. 203-207 
    Details
    ISSN: 1573-5060
    Keywords: barley ; electroporation ; PEG-mediated DNA uptake ; promoter analysis ; protoplasts ; transient expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary In some cereal species that are still recalcitrant to stable transformation and regeneration, transient expression in isolated protoplasts is a useful tool for the study of gene expression and regulation. We have successfully applied these techniques to barley protoplasts derived from developing endosperm, aleurone, leaves and roots in order to characterize functionally cis-acting motives in two gene promoters, corresponding to trypsin inhibitor BTI-CMe and to sucrose synthase Ss1. Gene specificity is maintained in transient expression assays with protoplasts isolated from these different barley tissues and the pattern of expression parallels the mRNA levels observed for the corresponding genes in the same tissues.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023949
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    Paper (German National Licenses)
    Fulltext
  • 3
    Electronic Resource
    Electronic Resource
    Transient gene expression in transformed banana (Musa cv. Bluggoe) protoplasts and embryogenic cell suspensions (1955)
    Springer
    Euphytica 85 (1955), S. 89-95 
    Details
    ISSN: 1573-5060
    Keywords: banana ; biolistic transformation ; electroporation ; embryogenic cell suspension ; Musa spp. ; protoplast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary In order to introduce currently-available genes with agronomical value into banana, two genetic transformation protocols have been optimized. Firstly, regenerable protoplasts isolated from embryogenic cell suspensions of the cultivar Bluggoe have been used for the introduction of several chimaeric uidA gene constructs by electroporation. With the inclusion of polyethylene glycol and heat shock, the frequency of transiently expressing protoplasts reached 1.8% as shown by an in situ β-glucuronidase assay. A duplicated 35S promoter with an alfalfa mosaic virus leader sequence (pBI-426) induced the highest expression rate among the constructs tested. Embryogenic cell suspensions of cv. Bluggoe have also been bombarded with accelerated particles coated with a high expression uidA gene construct (pEmuGN) using a biolistic gun. After a partial optimization of the procedure, transient GUS assays reproducibly demonstrated the presence of 400 blue foci in 30 μl of settled cell volume (approximately 25 mg cells). Selection and characterization of antibiotic-resistant transformed cultures is in progress.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023934
    Library Location Call Number Volume/Issue/Year Availability
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    Paper (German National Licenses)
    Fulltext
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