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  • 1
    Electronic Resource
    Electronic Resource
    Factors influencing transformation frequency of tomato (Lycopersicon esculentum) (1993)
    Springer
    Plant cell reports 12 (1993), S. 644-647 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We developed an efficient procedure for transformation and regeneration of L. esculentum cv. Moneymaker from cotyledon explants. The effect of two parameters on the transformation frequency was investigated in detail. The use of feeder layers during cocultivation proved to be critical. In addition, it was found that Agrobacterium strains harbouring a L,L-succinamopine type helper plasmid yielded significantly higher transformation frequencies than those with octopine or nopaline type helper plasmids. The optimized protocol was used to obtain transformation frequencies averaging 9%. Of the plants produced approximately 80% proved to be diploid, of which 67% contained the transgene(s) on a single locus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00232816
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  • 2
    Electronic Resource
    Electronic Resource
    Easy determination of ploidy level in Arabidopsis thaliana plants by means of pollen size measurement (1994)
    Springer
    Plant cell reports 13 (1994), S. 652-656 
    Details
    ISSN: 1432-203X
    Keywords: Transgenic Arabidopsis ; Ploidy ; Pollen size
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cytogenetic examination of transgenic Arabidopsis thaliana (L.) Heynh. plants obtained by Agrobacterium-mediated gene transfer to cotyledon- and root-explants or by direct gene transfer into protoplasts revealed a high percentage of tetraploid or aneuploid transformants. Depending on the transformation procedure used, 13% (root explant transformation), 33% (cotyledon explant transformation), or 38% (direct gene transfer) of the transformants showed aberrant ploidy levels. A good correlation between the ploidy level of a plant and the size of its pollen grains was observed. This allows quick and simple testing of the ploidy level of transgenic Arabidopsis plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00232939
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    Paper (German National Licenses)
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  • 3
    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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  • 4
    Electronic Resource
    Electronic Resource
    Regeneration of fertile plants from protoplasts of different Arabidopsis thaliana genotypes (1988)
    Springer
    Molecular genetics and genomics 213 (1988), S. 15-20 
    Details
    ISSN: 1617-4623
    Keywords: Arabidopsis thaliana ; Protoplast ; Plant regeneration ; Alginate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A protocol for obtaining regenerated fertile plants from mesophyll protoplasts of Arabidopsis thaliana is reported. Protoplasts were isolated from leaves of 21-to 28-day-old Arabidopsis plants grown in a controlled environment. Sustained divisions were achieved when protoplasts were embedded in beads formed by 1.4% sodium alginate in the presence of 50mM CaCl2 in 0.4 mannitol, which was then exchanged againts modified B5 medium. About 0.4%–0.6% of the protoplasts developed into colonies of which 80%–90% formed shoots and subsequently regenerated to fertile plants. Seeds harvested from more than 200 independently regenerated plants were sown and germination frequencies of more than 95% were obtained. Furthermore, the F1 plants did not show any evidence of somaclonal variation on visual inspection. This protocol was originally developed for Arabidopsis thaliana “Columbia”; however it was shown to be applicable also for the genotypes “Wassilewskija”, “Landsberg erecta” and “Estland” though with differing efficiencies.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00333392
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  • 5
    Electronic Resource
    Electronic Resource
    Efficient transformation of Arabidopsis thaliana using direct gene transfer to protoplasts (1989)
    Springer
    Molecular genetics and genomics 217 (1989), S. 6-12 
    Details
    ISSN: 1617-4623
    Keywords: Arabidopsis thaliana ; Protoplasts ; DNA transformation ; Polyethylene glycol ; Co-transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Direct gene transfer has proved to be an efficient transformation method for arabidopsis thaliana, a member of the Brassicaceae. Transgenic Arabidopsis plants resistant to hygromycin B have been regenerated from mesophyll protoplasts treated with polyethylene glycol and plasmid DNA carrying the hygromycin phosphotransferase (HPT) gene under the control of the 35 S promoter of cauliflower mosaic virus. The transformation procedure reproducibly yields transformants at frequencies of approximately 1×10-4 (based on the number of protoplasts treated) or 5% (based on the number of regenerating calli). DNA from plants regenerated from hygromycin resistant colonies was analysed by Southern blot hybridization demonstrating that the foreign gene is stably integrated into the plant chromosome. Genetic analysis of several hygromycin resistant plants showed that the HPT gene is transmitted to the progeny. Transformation experiments performed with a selectable and a non-selectable gene on separate plasmids resulted in a co-transformation rate of functionally active copies in about 25% of the transformants analysed. Hence this approach can be used to introduce non-selectable genes into the Arabidopsis genome.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00330935
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    Paper (German National Licenses)
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