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  • 1
    Electronic Resource
    Electronic Resource
    Electrofusion, a simple and reproducible technique in somatic hybridization of Nicotiana plumbaginifolia mutants (1985)
    Springer
    Plant cell reports 4 (1985), S. 274-276 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Electrofusion of protoplasts from two complementary nitrate reductase deficient mutants of Nicotiana plumbaginifolia has resulted in somatic hybrid lines. Mesophyll protoplasts isolated from the cofactor mutant CNX 20 and fluorescein diacetate stained protoplasts derived from a cell suspension culture of the NA 36 line, being defective in the apoenzyme, were used in the fusion experiments. In total, 594 lines were recovered which could proliferate on a selective medium with nitrate as the sole nitrogen source. This is including 141 putative hybrid lines which were obtained after transfer of 1048 heterokaryons with a micromanipulator one day after electrofusion. The hybrid character of some of the selected lines was confirmed by nitrate reductase activity measurements. Plants were grown from hybrid calli.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00269376
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  • 2
    Electronic Resource
    Electronic Resource
    Somatic hybrid potato plants after electrofusion of diploid Solanum tuberosum and Solanum phureja (1986)
    Springer
    Plant cell reports 5 (1986), S. 262-265 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts from diploid S. tuberosum and diploid S. phureja were electrofused followed by selection of the heterokaryons with a micromanipulator. Visual identification of the heterokaryons was facilitated by fluorescein diacetate staining of the protoplasts from one of the parents, which was grown on herbicide containing medium to induce bleaching of the chlorophyll. In total, 840 heterokaryons showing red (chlorophyll) and yellow-green (fluorescein) fluorescence were selected and cultured at relatively low densities using various feeder systems. Finally, 18 putative hybrid plantlets were obtained and grown to maturity. DNA histograms indicated that the plants were hexaploid, octoploid or mixoploid. With Giemsa C-band pattern analysis of the chromosomes the hybrid character and the combinations of the chromosome sets of all plants investigated could be established.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00269817
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  • 3
    Electronic Resource
    Electronic Resource
    Production of somatic hybrids between frost-tolerant Solanum commersonii and S. tuberosum: characterization of hybrid plants (1993)
    Springer
    Theoretical and applied genetics 87 (1993), S. 193-200 
    Details
    ISSN: 1432-2242
    Keywords: Frost tolerance ; Male sterility ; Potato ; Solanum commersonii ; Somatic hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Somatic fusion of mesophyll protoplasts was used to produce hybrids between the frost-tolerant species Solanum commersonii (2n=2x=24) and dihaploid S. tuberosum (2n=2x=24). This is a sexually incompatible combination due to the difference in EBN (Endosperm Balance Number, Johnston et al. 1980). Species with different EBNs as a rule are sexually incompatible. Fifty-seven hybrids were analysed for variation in chromosome number, morphological traits, fertility and frost tolerance. About 70% of the hybrids were tetraploid, and 30% hexaploid. Chloroplast counts in stomatal guard cells revealed a low frequency of cytochimeras. The frequency of aneuploids was relatively higher at the hexaploid level (hypohexaploids) than at the tetraploid level (hypotetraploids). The somatic hybrids were much more vigorous than the parents, and showed an intermediate phenotype for several morphological traits and moderate to profuse flowering. Hexaploid hybrid clones were less vigorous and had a lower degree of flowering than the tetraploid hybrid clones. All of the hybrids were female fertile but male sterile except for one, which was fully fertile and self-compatible. Many seeds were produced on the latter clone by selfing and on the male-sterile clones by crossing. The somatic hybrid plants showed an introgression of genes for frost tolerance and an adaptability to cold from S. commersonii. Therefore, the use of these somatic hybrids in breeding for and in genetic esearch on frost tolerance and cold-hardening is suggested.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00223764
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  • 4
    Electronic Resource
    Electronic Resource
    Tetraploid potato hybrids through protoplast fusions and analysis of their performance in the field (1992)
    Springer
    Theoretical and applied genetics 83 (1992), S. 807-812 
    Details
    ISSN: 1432-2242
    Keywords: Solanum tuberosum ; Solanum phureja ; Somatic hybrids ; Field experiment ; Agronomic traits
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Data on the production of tetraploid hybrid plants after electrofusion of protoplasts from various diploid (2n = 2x =24) Solarium tuberosum and S. phureja are reported. Ten different fusion combinations were used. Six hybrids with the tetraploid chromosome number 2n=4x=48 were tested under field conditions for their performance in various agronomic traits (tuber type, tuber yield, underwater weight of tubers, numbers of tubers, mean weight per tuber). Tuber yield in five of the six hybrid clones was similar to that of cv Bintje. One hybrid clone (35-4) had three times higher tuber yield than cv Bintje. The mean tuber weight of this hybrid was similar to that of cv Bintje. The results of this study prove that fertile tetraploid somatic hybrids having similar or higher tuber yield than that of cv Bintje can be obtained through somatic hybridization. This technique is now included in commercial potato breeding programs in The Netherlands.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00226701
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  • 5
    Electronic Resource
    Electronic Resource
    Nuclear genomic composition of asymmetric fusion products between irradiated transgenic Solanum brevidens and S. tuberosum: limited elimination of donor chromosomes and polyploidization of the recipient genome (1993)
    Springer
    Theoretical and applied genetics 86 (1993), S. 237-244 
    Details
    ISSN: 1432-2242
    Keywords: Solanum brevidens ; Solanum tuberosum ; Agrobacterium transformation ; Asymmetric somatic hybrids ; Restriction fragment length polymorphism (RFLP)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The production of asymmetric somatic hybrid calli after fusion between gamma-irradiated protoplasts from transgenic Solanum brevidens and protoplasts from S. tuberosum are reported. Transgenic (kanamycin-resistant, GUS-positive) S. brevidens plants and hairy root clones were obtained after transformation with Agrobacterium tumefaciens LBA 1060 (pRi1855) (pBI121) and LBA 4404 (pRAL4404) (pBI121), and A. rhizogenes LBA 9402 (pRi1855) (pBI121), respectively. Leaf protoplasts isolated from the transgenic plants or root protoplasts from the hairy root clones were fused with S. tuberosum leaf protoplasts, and several calli were selected on kanamycin-containing medium. The relative nuclear DNA content of the hybrid calli was measured by flow cytometry (FCM), and the percentages of DNA of the S. brevidens and S. tuberosum genomes in the calli were determined by dot blot analysis using species-specific DNA probes. Chromosome-specific restriction fragment length polymorphism (RFLP) markers were used to investigate the elimination of specific S. brevidens chromosomes in the hybrids. The combined data on FCM, dot blot and RFLP analysis revealed that 18–62% of the S. brevidens DNA was eliminated in the hybrid calli and that the RFLP marker for chromosome 7 was absent in seven out of ten calli. The absence of RFLP markers for chromosomes 5 and 11 hardly ever occurred. In most of the hybrids the ploidy level of the S. tuberosum genome had increased considerably.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00222085
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  • 6
    Electronic Resource
    Electronic Resource
    Elimination of Solanum phureja nucleolar chromosomes in S. tuberosum + S. phureja somatic hybrids (1987)
    Springer
    Theoretical and applied genetics 73 (1987), S. 878-882 
    Details
    ISSN: 1432-2242
    Keywords: Potatoes ; Somatic hybrid plants ; Nucleolar chromosomes ; Chromosome elimination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The karyotype of the dihaploid SVP1 line of S. tuberosum (2n=2x=24) showed two nucleolar chromosomes with differently sized satellites. The diploid SVP5 line (2n=2x=24) and tetraploid regenerants of S. phureja had larger but similar satellites. Somatic hybrids between the diploid lines of these potato species with genome combinations 4 tub + 2 ph (plants 1–3), 2 tub + 4 ph (plants 4–7) and 4 tub + 4 ph (plant 8) had lost 2 phureja nucleolar chromosomes if 4 phureja genomes were present. One phureja nucleolar chromosome of plants 1–3 and both of plants 5 and 7 had rearranged satellites. Elimination of the two nucleolar chromosomes occurred preferentially, was under genetic control, and probably took place during early callus development. NOR activity resulting in rear-rangements between NORs may have caused the elimination.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00289393
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  • 7
    Electronic Resource
    Electronic Resource
    Some methodological aspects of apple transformation by Agrobacterium (1955)
    Springer
    Euphytica 85 (1955), S. 131-134 
    Details
    ISSN: 1573-5060
    Keywords: apple ; transformation ; Agrobacterium ; preculture ; azacytidine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Leaf explants of apple cvs Gala and Golden Delicious were infected with the Agrobacterium tumefaciens strain AGL0(pMOG410). The effects of a 2 d preculture of the explants before infection and the addition of 5-azacytidine to the selection medium were studied. The percentages of GUS-positive explants after 5 w did not significantly alter due to these treatments. One of the ‘Gala’ shoots, which was removed from a leaf explant cultured for 8 w on selection medium, proved to be GUS-positive and will be analyzed further. In general, however, it should be concluded that regeneration of transgenic shoots directly from leaf tissue was not very effective.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023941
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