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  • protoplasts  (7)
  • shoot regeneration  (3)
  • 1
    ISSN: 1432-203X
    Schlagwort(e): Vigna sublobata ; protoplasts ; microcalli ; shoot bud formation ; plant regeneration
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Viable protoplasts of Vigna sublobata L. were isolated enzymatically from hypocotyls of axenic seedlings. Protoplast yields were dependent upon seedling age, with maximum yields (2.25 ± 0.35 × 106 g fwt−1) from seedlings aged 6 d. Protoplasts regenerated cell walls and underwent sustained divisions when cultured in either agarose-solidified or liquid K8P medium. The plating density affected the division frequency and plating efficiency; the division frequency (68 ±0 6.0%) was maximum at 4.0 × 104 ml−1 while plating efficiency was maximum (1.3 ± 0.1%) at 5.0 × 104 ml−1. Dividing protoplasts developed into microcalli, which produced glossy green compact nodular calli on transfer to 8.0 gl−1 w/v agar-solidified medium containing MS salts, B5 organic components, 30 g l−1 sucrose, NAA (0.2–0.5 mg l−1), zeatin riboside (0.5–2.0 mg l−1) and GA3 (0.5–1.0 mg l−1). These calli, after sub-culture on the same medium, produced shoot buds which underwent elongation following transfer of tissues to 6.0 g l−1 agar-solidified B5 medium containing 30g l−1 sucrose, IBA (0.01 mg l−1) and BAP (1.0 mg l−1). Elongated shoots developed roots after transfer to 8.0g l−1 agar-solidified, hormone-free MS medium with 30 g l−1 sucrose.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    ISSN: 1432-203X
    Schlagwort(e): Corchorus capsularis ; jute ; cotyledons ; surfactants ; Pluronic F-68 ; Triton X-100 ; Tween 20 ; shoot regeneration
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary The effects have been studied of the non-ionic surfactants, Plutonic F-68, Tween 20 or Triton X-100, on shoot regeneration from cultured jute (Corchorus capsularis L.) cotyledons with attached petioles. This group of non-ionic surfactants was selected in order to determine a possible relationship between the physico-chemical properties of individual compounds and their observable effects on plant morphogenesis in cultured jute cotyledons. Supplementation of culture medium with 0.001–0.5% (w/v) Pluronic F-68 increased the mean percentage of cotyledons producing shoots and the mean number of shoots/cotyledon, with maximal responses at 0.5% (w/v). By contrast, Tween 20 produced maximal effects at 0.001% (v/v), with inhibition of shoot formation at 0.5% (v/v). In both cases, phenotypically normal plants were recovered which could be grown to maturity. Culture of cotyledons with 0.001% (v/v) Triton X-100 similarly increased both the percentage of cotyledons producing shoots and the number of shoots/cotyledon. However, these shoots did not develop into mature plants. Additionally, shoots did not regenerate from cotyledons cultured with Triton at 0.01–0.5% (v/v). These results demonstrate mat there is an apparent relationship between the hydrophilic-hydrophobic (HLB) balance of surfactants which determine their cell permeabilising properties and consequently, their effects on morphogenesis.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 3
    ISSN: 1432-203X
    Schlagwort(e): Petunia hybrida ; protoplasts ; oxygen delivery ; perfluorochemicals ; Pluronic F-68 ; surfactant ; cell division
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Cell suspension-derived protoplasts of albino Petunia hybrida were grown for 10 d at the interface between aqueous culture medium (KM8P) and an oxygenated (10 mbar for 15 min) perfluorocarbon liquid, perfluorodecalin. Protoplasts synthesised new cell walls and divided normally at the perfluorodecalin/culture medium interface, with a mean viability after 10 d of 〉 92.0%. The mean plating efficiency of protoplasts was elevated by 37% (P〈0.05) following culture at the perfluorodecalin/medium interface, but was unaltered by perfluorodecalin or oxygen separately. The mean plating efficiency of protoplasts cultured at the interface was further increased to a maximium of 52% above control, in the presence of oxygenated perfluorodecalin and KM8P medium supplemented with the non-ionic, co-polymer surfactant, Pluronic F-68 at 0.01% (w/v). These findings demonstrate the effectiveness of oxygenated perfluorodecalin for promoting protoplast growth, by facilitating oxygen delivery. The finding that Pluronic F-68 further increased the plating efficiency of protoplasts cultured at the perfluorocarbon/aqueous interface suggests that these agents improve growth through separate, but cumulative, mechanisms.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 4
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell, tissue and organ culture 32 (1993), S. 311-317 
    ISSN: 1573-5044
    Schlagwort(e): cell suspension ; Lycopersicon chilense ; plant regeneration ; protoplasts ; wild tomato
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A protocol has been established for rapid, high frequency plant regeneration from protoplasts of the wild tomato species Lycopersicon chilense Dun. Cell suspension cultures were obtained from calli initiated from seedling stem explants. Protoplasts were isolated from cell suspensions by an overnight one-step enzyme digestion, purified by washing in salts solution and cultured in liquid medium. Dilution of liquid medium every 3 days, with medium containing low levels of growth regulators and sucrose, was critical for sustained colony formation. Up to 70% of protoplast-derived calli regenerated shoots when cultured on agar-solidified medium with Murashige & Skoog (1962) salts and vitamins, 2.0 mg l-1 zeatin and 0.1 mg l-1 indole acetic acid for 21 days, followed by transfer to the same medium lacking indole acetic acid.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 5
    ISSN: 1573-5044
    Schlagwort(e): Hordeum vulgare ; plasmid uptake ; polyethylene glycol (PEG) ; protoplasts ; transient gene expression ; stable transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Cat gene expression has been investigated following PEG-mediated plasmid uptake into barley protoplasts. The uptake conditions optimised for transient expression were employed for stable transformation. Transformed protoplast-derived calli of the cvs. Dissa and Igri, were selected on medium containing G418 at 40 μg ml−1 or kanamycin sulphate at 250 μg ml−1. Absolute transformation frequencies of 28.9×10−5 and 21.3×10−5 were recorded for Dissa with kanamycin sulphate and G418 selection, respectively. The frequency for Igri was 11.5×10−5 with G418 selection. Antibiotic resistant protoplast-derived colonies expressed NPTII activity; Southern hybridisation confirmed integration of the nptII gene into barley genomic DNA.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 6
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell, tissue and organ culture 36 (1994), S. 255-258 
    ISSN: 1573-5044
    Schlagwort(e): tomato ; protoplasts ; plant regeneration
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Three Indian cultivars ofL. esculentum were assessed for shoot regeneration from protoplast-derived calli. Consistent yields of viable protoplasts (〉9.0×106 g f.wt.-1) were obtained from leaflets of 14 days old cultured shoots. Protoplast viability (88–94%) and planting efficiency (55–70%) were recorded for the three cultivars. Up to 71% of the protoplast-derived tissues regenerated shoots.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 7
    Digitale Medien
    Digitale Medien
    Springer
    Euphytica 102 (1998), S. 195-202 
    ISSN: 1573-5060
    Schlagwort(e): cotyledonary node ; Phaseolus vulgaris ; P. coccineus ; shoot regeneration
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Phaseolus vulgaris L. is the most important economic species within the genus Phaseolus, and it is grown in all parts of the world. Genetic improvement by conventional breeding has met considerable success, although production of hybrids between species within the genus has been limited due to sexual incompatibility or other evolutionary lethalities. Recent advances in tissue culture have offered the opportunity to produce cultivars which could not be obtained by conventional breeding methods, but regeneration protocols are influenced by the genotype. A standard regeneration procedure was assessed for its applicability to elite breeding lines of P. vulgaris L. and landraces of P. coccineus L. from seedling explants containing a cotyledon and a small portion of the split embryonic axis. In vitro culture response and regeneration ability varied significantly between species and amongst genotypes. P. coccineus produced more shoots per explant with a higher rooting efficiency than P. vulgaris. These significant genotype effects suggest that genetic factors are important in the response to in vitro tissue culture. Different genotypes were identified, which were more competent to in vitro culture and could produce highly responsible hybrids. This in vitro culture system carry a high potential for propagation of P. vulgaris and P. coccineus, and subsequent exploitation of hybrid forms, which could also be incorporated into somatic cell approaches to improve these species.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 8
    ISSN: 1573-5060
    Schlagwort(e): Oryza sativa L. cv. Taipei 309 ; rice ; protoplasts ; direct DNA uptake ; kanamycin-resistant transgenic plants ; field trial ; glasshouse trial ; neomycin phosphotransferase II (npt II) gene ; gene expression and inheritance
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary The phenotypes of seed progeny (R2 generation) of Oryza sativa L. cv. Taipei 309, which carried the neomycin phosphotransferase II (npt II) gene, were compared with those of non-transformed, protoplast-derived plants of the same generation and non-transformed, seed-derived plants under field and glasshouse conditions. Under both conditions the transgenic plants were generally smaller, took longer to flower and had reduced fertility. Significant differences were observed between individuals within the group of transgenic plants. The npt II gene was present in most of the transgenic plants, but NPT II activity was only detected in a minority of individuals.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 9
    Digitale Medien
    Digitale Medien
    Springer
    Plant growth regulation 17 (1995), S. 233-239 
    ISSN: 1573-5087
    Schlagwort(e): micropropagation ; shoot regeneration ; Pluronic F-68 ; Poplar
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Stem explants and leaves (without petioles) excised from axenic shoots of Populus tremula cv. Ahle or P. tremula × tremuloides cv. Münden were cultured in the presence of the non-ionic, co-polymer surfactant, Pluronic F-68. Stem explants developed shoots within 10 d of culture and significant (p〈0.05), but genotype-dependent, increases in total shoot fresh weight (maximum 2 × control) occurred in cultures supplemented with 0.001–0.1% Pluronic F-68 over a 72 d period. Similarly, increases in both fresh weight (up to 10-fold) and number of shoots per P. tremula × tremuloides leaf explant (5-fold maximum) over 60 d occurred with Pluronic F-68 at 0.001%.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 10
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell, tissue and organ culture 30 (1992), S. 69-75 
    ISSN: 1573-5044
    Schlagwort(e): chloramphenicol acetyltransferase ; electroporation ; Eucalyptus citriodora ; protoplasts ; transient gene expression
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Protoplasts isolated from cotyledons of Eucalyptus citriodora were electroporated using a rectangular pulse, with plasmid carrying the cat gene. The levels of transient expression and protoplast viability were influenced by the voltage and pulse duration. At a field strength of 800 V cm-1 (1000 μs), a protoplast viability of 57%, and 47% conversion of 14C-chloramphenicol to its acetylated forms, were obtained. Expression levels were improved by an increase in plasmid concentration (up to 60 μg ml-1), and also by the addition of carrier DNA. Gene expression was further enhanced by the addition of 40% (w/v) PEG, in the presence of the carrier DNA, to the protoplasts after electroporation.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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