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  • Agrobacterium  (2)
  • Transformation  (2)
  • 1
    ISSN: 1432-203X
    Keywords: Brassica oleracea ; Direct gene transfer ; Protoplast regeneration ; Transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Efficient regeneration (80%) and high frequency genetic transformation (10–33%) were achieved by culturing protoplasts isolated from hypocotyl tissues of six day old Brassica oleracea seedlings and by subjecting these protoplasts to PEG mediated direct plasmid uptake. Three different plasmid vectors carrying marker genes for resistance to methotrexate (dhfr), hygromycin (hpt) and phosphinotricin (bar) were constructed and used for transformation. Large number of normal, fertile transformants were obtained with vectors carrying hpt and bar genes. No transformants could be regenerated for resistance to methotrexate as it severely suppressed shoot differentiation.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5028
    Keywords: Agrobacterium ; growth retardants ; plant hormones ; Ri plasmid ; transgenic tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The rolA gene of the TL-DNA of Agrobacterium rhizogenes Ri-plasmid plays a major role in establishing the hairy root syndrome in transgenic plants. Transgenic tobacco plants (Nicotiana tabacum L.) expressing constitutively the rolA gene under the transcriptional control of the 35S RNA promoter show pronounced phenotypical alterations. P35S-rolA transgenic tobacco plants are characterized by stunted growth, dark green wrinkled leaves with an altered length-to-width ratio, condensed inflorescences, retarded onset of flowering, a reduced number of flowers and shortened styles. To investigate whether the pleiotropic alterations of growth and development are linked to an altered hormonal status we have compared the immunoreactive content of indole-3-acetic acid, cytokinins, abscisic acid, gibberellin and the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) of seedlings and different tissues of P35S-rolA transgenic plants, transgenic plants expressing the rolA gene under control of its own phloem-specific promoter and wild-type plants. Multiple tissue-specific alterations of phytohormone concentrations are the consequence of rolA gene activity. Changes of phytohormonal content can explain part of the rolA-induced phenotypic alterations. Most strikingly, in young and fully developed leaves of rolA and P35S-rolA transgenic clones a 40–60% reduction of immunoreactive gibberellin A1 was found, as compared to wild-type leaves. Treatment of wild-type tobacco plants with inhibitors of gibberellin biosynthesis phenotypic alterations similar to those of rolA transgenic plants. This suggests that the reduction of gibberellic acid content is indirectly but causally involved in rolA-induced alterations of stem elongation and planar leaf blade growth.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 219 (1989), S. 209-216 
    ISSN: 1617-4623
    Keywords: Agrobacterium tumefaciens ; Transformation ; Leaf disc infection ; Nicotiana tabacum ; TR promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The physiological function in planta of T-DNA gene 6b was studied under various experimental conditions. For this purpose the coding region of gene 6b was cloned behind the 1′-promoter of the TR-DNA to enhance expression of the gene product in transformed plant cells. Expression of the recombinant gene in leaf discs of Nicotiana tabacum altered the capacity for shoot formation of the discs, induced by exogenous (i.e. BAP in the growth medium or agrobacterial trans-zeatin produced under control of gene tzs) or endogenous cytokinins (i.e. isopentenyladenosine produced under control of T-DNA gene 4). The data obtained indicate a reduction of cytokinin activity within the plant cells by the product of T-DNA gene 6b.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1617-4623
    Keywords: Agrobacterium ; Light induction ; Position effect ; Transformed plants ; Gene isolation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The isolation and analysis of several cDNAs and of one genomic clone encoding ST-LS1, a single-copy gene fromSolanum tuberosum with leaf/stem-specific, light-inducible expression, is described. The structure of the gene was determined by sequencing several overlapping partial cDNA clones as well as the genomic clone and by determining the transcription start site by RNA protection experiments. A “tagged” derivative of the gene, obtained by exon modification, was reintroduced into potato and into tobacco shoots usingAgrobacterium/Ti-plasmid vector systems. The modified gene was expressed in both tobacco and potato shoots giving rise to an RNA of approximately 1,200 nucleotides which exceeded the length of the RNA made from the endogenous gene by the expected size of the “tag” (470 nucleotides). The level of expression of the modified gene varied substantially between independent transformants. A high proportion of the transformants (20%–40%) synthesized as much RNA from the added gene as from the resident gene. Expression of the transferred gene was light induced. Qualitatively and quantitatively the expression of the introduced gene was similar in a homologous (potato) and in a heterologous, but related, cellular background (tobacco).
    Type of Medium: Electronic Resource
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