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  • 1
    Electronic Resource
    Electronic Resource
    Analysis of DNA polymerase activity in Petunia protoplasts treated with clastogenic agents (1994)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 90 (1994), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Clastogenic agents, i.e. agents that can induce chromosome or DNA breakage, have been shown to enhance the rale of direct gene transfer to protoplasts. The effect was analysed at the enzymatic level using protoplast homogenates as well as intact protoplasts. For that purpose existing procedures were modified to enable measurement of DNA polymerase in vivo. In the system used, external DNA was able to enter the cells without the addition of membrane-permeabilizing compounds. When comparing total DNA polymerase activity of protoplasts irradiated with X-rays or UV-light with that of untreated cells we did not observe significant differences. Incubation of protoplasts with high doses of bleomycin affected total DNA polymerase activity negatively. but dideoxythymidine triphosphate-sensitive activity was not influenced. We conclude that the DNA strand-breaks induced by low doses of X-rays. UV-light or bleomycin do not increase the total or the repair-DNA polymerase activity and. therefore. that the increase in the transformation rates after DNA strand-breaking is not preceded by enhanced DNA polymerase activity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1994.tb08800.x
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  • 2
    Electronic Resource
    Electronic Resource
    Plant regeneration from protoplasts of sugar beet (Beta vulgaris) (1995)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 94 (1995), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Mesophyll protoplasts from two of five sugar beet lines tested were regenerated into plants. Mesophyll protoplasts of all lines showed high plating efficiencies up to 4.0% developed hard compact callus, and two of the lines also developed white, soft and friable callus consisting of starch grain-containing cells. Whereas the compact callus never regenerated into plants, the white friable ones frequently developed globular structures, which became green in the light and formed adventitious shoots after cytokinin (BAP or thidiazuron) treatment. Genetic analysis by PCR-fingerprinting and flow cytometry showed uniformity and unchanged ploidy levels in 15 independently regenerated plantlets in line NF. but altered ploidy level (from diploid to triploid) in a regenerated plantlet of clone VRB.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1995.tb05321.x
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  • 3
    Electronic Resource
    Electronic Resource
    Foreign DNA sequences are received by a wild-type strain of Aspergillus niger after co-culture with transgenic higher plants (1994)
    Springer
    Current genetics 27 (1994), S. 70-76 
    Details
    ISSN: 1432-0983
    Keywords: Hygromycin resistance ; Expression ; Aspergillus niger ; Gene transfer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Different transgenic plants of Brassica napus, Brassica nigra, Datura innoxia and Vicia narbonensis expressing the hph gene under the control of the 35s promoter were co-cultivated with mycelial material of Aspergillus niger in microcosms under sterile conditions. A significantly higher number of hygromycin B-resistant colonies of re-isolated fungi was obtained if compared with co-cultures with non-transgenic plants. The hph gene and other foreign sequences could be detected in some of the resistant strains only for a short time after selection, indicating a rapid loss of foreign DNA. A more stable transgenic strain was obtained after co-culture with transgenic plants of D. innoxia including a high number of hph copies in their genome. DNA with detected pUC sequences was prepared to transform E. coli DH5α. One of the recovered plasmids is shown to include pieces of the plant-transforming vector and a foreign sequence. The 35s-regulated expression of genes is studied in A. niger.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00326581
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  • 4
    Electronic Resource
    Electronic Resource
    Plant regeneration from protoplasts ofVicia narbonensis via somatic embryogenesis and shoot organogenesis (1996)
    Springer
    Plant cell reports 16 (1996), S. 22-25 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts ofVicia narbonensis isolated from epicotyls and shoot tips of etiolated seedlings were embedded in 1.4% sodium-alginate at a final density of 2.5×105 protoplasts/ml and cultivated in Kao and Michayluk-medium containing 0.5 mg/I of each of 2,4- dichlorophenoxyacetic acid, naphthylacetic acid and 6 -benzylaminopurine. A division frequency of 36% and a plating efficiency of 0.40–0.5% were obtained. Six weeks after embedding, protoplast-derived calluses were transferred onto gelrite-solidified Murashige and Skoog-media containing various growth regulators. Regeneration of plants was achieved via two morphologically distinguishable pathways. A two step protocol (initially on medium with a high auxin concentration followed by a culture phase with lowered auxin amount) was used to regenerate somatic embryos, whereas cultivation on medium containing thidiazuron and naphthylacetic acid resulted in shoot morphogenesis. Mature plants were recovered from both somatic embryos as well as from thidiazuron-induced shoots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01275442
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  • 5
    Electronic Resource
    Electronic Resource
    Genetic evidence for the hybrid nature of somatic hybrids from Datura innoxia Mill. (1978)
    Springer
    Planta 141 (1978), S. 333-334 
    Details
    ISSN: 1432-2048
    Keywords: Anther culture ; Datura ; Somatic hybrids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The hybrid nature of tetraploid somatic hybrids of two genetically different chlorophyll-deficient mutants from Datura innoxia Mill. was demonstrated with the aid of anther culture. Green and chlorophyll-deficient androgenetic lines could be regenerated from the pollen grains.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00388353
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  • 6
    Electronic Resource
    Electronic Resource
    Transformation of haploid Datura innoxia protoplasts and analysis of the plasmid integration pattern in regenerated transgenic plants (1993)
    Springer
    Plant cell reports 12 (1993), S. 390-394 
    Details
    ISSN: 1432-203X
    Keywords: Datura ; haploids ; direct gene transfer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We developed a highly efficient transformation protocol for the PEG-mediated direct transfer of plasmid DNA into protoplasts of haploid Datura innoxia. Vectors harbouring a neomycin phosphotransferase II gene or a hygromycin B phosphotransferase gene under the control of different promoters were used in the transformation experiments. Various amounts of plasmid DNA were applied without any carrier DNA to show the direct influence of the plasmid DNA concentration on the transformation efficiency. Approximately 95% of the selected calli were regenerated to plants; 20% of them remained haploid. Total DNA of different transgenic plants was analysed with regard to the integration pattern of the plasmid DNA. Plants carrying only one or two copies of the vector DNA were observed as well as individuals with multi-copy integration (up to ten or more copies).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00234698
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  • 7
    Electronic Resource
    Electronic Resource
    Thidiazuron-induced plant regeneration from protoplasts of Vicia faba cv. Mythos (1995)
    Springer
    Plant cell reports 15 (1995), S. 164-169 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Protoplasts of 10 cultivars of V. faba were isolated from etiolated shoot-tips and tested for their regeneration capacity. After purification, protoplasts were embedded in sodium alginate and cultivated in the medium of Kao and Michayluk (1975) containing 0.5 mg·1−1 of each 2,4-dichlorophenoxyacetic acid, naphthylacetic acid and 6-benzylaminopurine. Depending on cultivar, division frequencies of up to 40% were obtained. Six weeks after embedding, protoplast-derived calluses were transferred to Gelrite-solidified media with different combinations of growth regulators. A two step protocol (auxin high/low) was tested for its ability to induce somatic embryogenesis. The formation of globular structures was observed, but no embryo formation could be achieved. In contrast, cultivation of protocalluses on medium supplemented with thidiazuron resulted in shoot development in cultivar Mythos. To generate mature plants, the shoots were grafted onto young seedlings. In order to optimize the in vitro-conditions, different concentrations of thidiazuron alone or in combination with naphthylacetic acid were tested, showing that an increase of thidiazuron and the addition of naphthylacetic acid positively affects both the viability of protocalluses and the regeneration frequency.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00193712
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  • 8
    Electronic Resource
    Electronic Resource
    Über das Chemotaktikum des Lebermooses (1971)
    Springer
    Naturwissenschaften 58 (1971), S. 568-568 
    Details
    ISSN: 1432-1904
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00598724
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  • 9
    Electronic Resource
    Electronic Resource
    Plant regeneration from protoplasts of Vicia narbonensis via somatic embryogenesis and shoot organogenesis (1996)
    Springer
    Plant cell reports 16 (1996), S. 22-25 
    Details
    ISSN: 1432-203X
    Keywords: Abbreviations:BAP: 2,4-D, 2,4-dichlorophenoxyacetic acid; BA, 6-benzylaminopurine; CH, casein hydrolysate; FDA, fluorescein diacetate; IBA, indole-3-butyric acid; KM, Kao and Michayluk's medium (1975); MS, Murashige and Skoog's medium (1962); NAA, maphthylacetic acid; TDZ, thidiazuron
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts of Vicia narbonensis isolated from epicotyls and shoot tips of etiolated seedlings were embedded in 1.4% sodium-alginate at a final density of 2.5×105 protoplasts/ml and cultivated in Kao and Michayluk-medium containing 0.5 mg/l of each of 2,4-dichlorophenoxyacetic acid, naphthylacetic acid and 6-benzylaminopurine. A division frequency of 36% and a plating efficiency of 0.4–0.5% were obtained: Six weeks after embedding, protoplast-derived calluses were transferred onto gelrite-solidified Murashige and Skoog-media containing various growth regulators. Regeneration of plants was achieved via two morphologically distinguishable pathways. A two step protocol (initially on medium with a high auxin concentration followed by a culture phase with lowered auxin amount) was used to regenerate somatic embryos, whereas cultivation on medium containing thidiazuron and naphthylacetic acid resulted in shoot morphogenesis. Mature plants were recovered from both somatic embryos as well as from thidiazuron-induced shoots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002990050168
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  • 10
    Electronic Resource
    Electronic Resource
    Transformation of Vicia narbonensis via Agrobacterium-mediated gene transfer (1991)
    Springer
    Plant cell reports 9 (1991), S. 535-538 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Shoot tips and epicotyl-segments of Vicia narbonensis were co-cultivated with Agrobacterium tumefaciens strain C58C1 pGV 3850 HPT, carrying a plasmid coding for hygromycin-phosphotransferase. On callus-induction medium containing 60 mg/l hygromycin for selection, approximately 18% of the explants produced hygromycin-resistant callus. After transfer to regeneration-medium these calluses produced hygromycin-resistant and nopaline-positive somatic embryos which could be regenerated to plantlets. The integration of the T-DNA into the plant genome was confirmed by Southern analysis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00232326
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