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  • 1
    Electronic Resource
    Electronic Resource
    Expression and inheritance of inserted markers in binary vector carrying Agrobacterium rhizogenes-transformed potato (Solanum tuberosum L.) (1989)
    Springer
    Theoretical and applied genetics 78 (1989), S. 705-714 
    Details
    ISSN: 1432-2242
    Keywords: Agrobacterium rhizogenes ; Solanum tuberosum ; Binary vector ; Neomycin phosphotransferase ; β-Glucuronidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transgenic shoots were regenerated from eight diploid potato hairy root clones obtained by transformation with Agrobacterium rhizogenes harboring next to its wild-type Ri-plasmid a binary vector containing the neomycin phosphotransferase and the β-glucuronidase genes. The plants exhibited the typical hairy root phenotype. Of the plants isolated, 58% were tetraploid and 38% were diploid. Flowering and tuberization was much better in the diploid than in the tetraploid plants. Transgenic plants formed a significantly larger root system when grown on kanamycin-containing medium as compared to growth on kanamycin-free medium. Direct evidence for genetic transformation was obtained by opine, neomycin phosphotransferase and β-glucuronidase assays, and by molecular hybridization. Fourteen flowering diploid plants were reciprocally crossed with untransformed S. tuberosum plants, but only six were successful. Seedlings obtained from four crosses showed that all traits were transmitted to the offspring. Molecular analysis confirmed the presence of multiple integrations (copies) of both vector T-DNA and Ri-T-DNA. The genetic data, furthermore, suggest that the traits derived from Ri-T-DNA and binary vector T-DNA are linked, as no recombination between the different traits was observed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00262567
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  • 2
    Electronic Resource
    Electronic Resource
    Isolation of an amylose-free starch mutant of the potato (Solanum tuberosum L.) (1987)
    Springer
    Theoretical and applied genetics 75 (1987), S. 217-221 
    Details
    ISSN: 1432-2242
    Keywords: Solanum tuberosum ; Mutant ; Starch composition ; Granule-bound starch synthase ; amylose-free
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An amylose-free potato mutant was isolated after screening 12,000 minitubers. These minitubers had been induced on stem segments of adventitious shoots, which had been regenerated on leaf explants of a monoploid potato clone after Röntgen-irradiation. The mutant character is also expressed in subterranean tubers and in microspores. Starch granules from the mutant showed a strongly reduced activity of the granule bound starch synthase and loss of the major 60 kd protein from the starch granules.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00249167
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  • 3
    Electronic Resource
    Electronic Resource
    Expression of a wild-type GBSS gene introduced into an amylose-free potato mutant by Agrobacterium tumefaciens and the inheritance of the inserts at the microsporic level (1994)
    Springer
    Theoretical and applied genetics 88 (1994), S. 369-375 
    Details
    ISSN: 1432-2242
    Keywords: Complementation ; Granule-bound starch synthase ; Amylose content ; Gene expression ; Inheritance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Granule-bound starch synthase (GBSS) catalyses the synthesis of amylose in starch granules. Transformation of a diploid amylose-free (amf) potato mutant with the gene encoding GBSS leads to the restoration of amylose synthesis. Transformants were obtained which had wild-type levels of both GBSS activity and amylose content. It proved to be difficult to increase the amylose content above that of the wild-type potato by the introduction of additional copies of the wild-type GBSS gene. Staining of starch with iodine was suitable for investigating the degree of expression of the inserted GBSS gene in transgenic amf plants. Of the 19 investigated transformants, four had only red-staining starch in tubers indicating that no complementation of the amf mutation had occured. Fifteen complemented transformants had only blue-staining starch in tubers or tubers of different staining categories (blue, mixed and red), caused either by full or partial expression of the inserted gene. Complementation was also found in the microspores. The segregation of blue- and red-staining microspores was used to analyse the inheritance of the introduced GBSS genes. A comparison of the results from microspore staining and Southern hybridisation indicated that, in three tetraploid transgenics, the gene was probably inserted before (duplex), and in all others after, chromosome doubling (simplex). The partial complementation was not due to methylation of the HPAII/MSPI site in the promoter region. Partially complemented plants had low levels of mRNA as was found when the GBSS expression levels were inhibited by anti-sense technology.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00223647
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  • 4
    Electronic Resource
    Electronic Resource
    Efficient transformation of potato (Solanum tuberosum L.) using a binary vector in Agrobacterium rhizogenes (1989)
    Springer
    Theoretical and applied genetics 78 (1989), S. 594-600 
    Details
    ISSN: 1432-2242
    Keywords: Agrobacterium rhizogenes ; Solanum tuberosum ; Binary vector ; Neomycin phosphotransferase II ; β-Glucuronidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We transformed three potato (Solanum tuberosum L.) genotypes by using A. rhizogenes or a mixture of A. rhizogenes and A. tumefaciens. Inoculations of potato stem segments were performed with Agrobacterium rhizogenes AM8703 containing two independent plasmids: the wild-type Ri-plasmid, pRI1855, and the binary vector plasmid, pBI121. In mixed inoculation experiments, Agrobacterium rhizogenes LBA1334 (pRI1855) and Agrobacterium tumefaciens AM8706 containing the disarmed Ti-plasmid (pAL4404) and the binary vector plasmid (pBI121) were mixed in a 1∶1 ratio. The T-DNA of the binary vector plasmid pBI121 contained two marker genes encoding neomycin phosphotransferase, which confers resistance to kanamycin, and β-glucuronidase. Both transformation procedures gave rise to hairy roots on potato stem segments within 2 weeks. With both procedures it was possible to obtain transformed hairy roots, able to grow on kanamycin and possessing β-glucuronidase activity, without selection pressure. The efficiency of the A. rhizogenes AM8703 transformation, however, was much higher than that of the “mixed” transformation. Up to 60% of the hairy roots resulting from the former transformation method were kanamycin resistant and possessed β-glucuronidase activity. There was no correlation between the height of the kanamycin resistance and that of the β-glucuronidase activity in a root clone. Hairy roots obtained from a diploid potato genotype turned out to be diploid in 80% of the cases. Transformed potato plants were recovered from Agrobacterium rhizogenes AM8703-induced hairy roots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00290847
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  • 5
    Electronic Resource
    Electronic Resource
    Inhibition of the expression of the gene for granule-bound starch synthase in potato by antisense constructs (1991)
    Springer
    Molecular genetics and genomics 225 (1991), S. 289-296 
    Details
    ISSN: 1617-4623
    Keywords: Amylose content ; Antisense RNA ; Dominant (hemizygous) suppression ; Granule-bound starch synthase ; Transgenic potato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Granule-bound starch synthase [GBSS; EC 24.1.21] determines the presence of amylose in reserve starches. Potato plants were transformed to produce antisense RNA from a gene construct containing a full-length granule-bound starch synthase cDNA in reverse orientation, fused between the cauliflower mosaic virus 35S promoter and the nopaline synthase terminator. The construct was integrated into the potato genome by Agrobacterium rhizogenes-mediated transformation. Inhibition of GBSS activity in potato tuber starch was found to vary from 70% to 100%. In those cases where total suppression of GBSS activity was found both GBSS protein and amylose were absent, giving rise to tubers containing amylose-free starch. The variable response of the transformed plants indicates that position effects on the integrated sequences might be important. The results clearly demonstrate that in tubers of potato plants which constitutively synthesize antisense RNA the starch composition is altered.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00269861
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    Paper (German National Licenses)
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