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  • Electronic Resource  (11)
  • 1990-1994  (11)
  • 1955-1959
  • 1850-1859
  • 1994  (11)
  • protoplasts
  • 1
    Electronic Resource
    Electronic Resource
    Isolation and transformation of rice aleurone protoplasts (1994)
    Springer
    Plant cell reports 13 (1994), S. 394-396 
    Details
    ISSN: 1432-203X
    Keywords: Rice ; α-amylase ; protoplasts ; aleurone ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Protoplasts isolated from the aleurone have been used extensively in molecular studies focusing on hormone-mediated regulation of gene expression in barley seed. To extend the use of aleurone protoplasts to other species, we have determined the conditions necessary for the isolation of protoplasts from rice aleurone layers of germinated seed. Many of the common cell wall degrading enzymes used in making protoplasts were tested for their ability to release protoplasts from rice aleurone layers. Cellulysin was found to be the most effective. Transformation of these aleurone protoplasts was accomplished using polyethylene glycol and DNA constructs containing the firefly luciferase reporter gene under the control of two different promoters were tested. Luciferase expression was 24-fold greater when the reporter gene was under the control of the CaMV 35S promoter than when the promoter from the alcohol dehydrogenase 1 gene was used. With the isolation and transformation of aleurone protoplasts from rice, it is now possible to investigate molecular events occurring in this tissue during germination.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00234145
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  • 2
    Electronic Resource
    Electronic Resource
    A reproducible procedure for plant regeneration from seedling hypocotyl protoplasts of Vigna sublobata L. (1994)
    Springer
    Plant cell reports 14 (1994), S. 175-179 
    Details
    ISSN: 1432-203X
    Keywords: Vigna sublobata ; protoplasts ; microcalli ; shoot bud formation ; plant regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: 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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233785
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  • 3
    Electronic Resource
    Electronic Resource
    Synergistic enhancement of protoplast growth by oxygenated perfluorocarbon and Pluronic F-68 (1994)
    Springer
    Plant cell reports 13 (1994), S. 251-255 
    Details
    ISSN: 1432-203X
    Keywords: Petunia hybrida ; protoplasts ; oxygen delivery ; perfluorochemicals ; Pluronic F-68 ; surfactant ; cell division
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: 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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233314
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  • 4
    Electronic Resource
    Electronic Resource
    The biosynthetic pathway of the S-alk(en)yl-L-cysteine sulphoxides (flavour precursors) in species of Allium (1994)
    Springer
    Plant cell, tissue and organ culture 38 (1994), S. 181-188 
    Details
    ISSN: 1573-5044
    Keywords: Allium cepa ; biosynthesis ; compartmentation ; γ-glutamyl peptides ; onion ; protoplasts ; S-alkenyl-L-cysteine sulphoxide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Pulse labelling experiments with 35SO4 2- fed for 24h to intact plants (shooted onion sets)of Allium cepa (onion) showed that 〉70% of the label appeared in the S-alkenyl-L-cysteine sulphoxides within 18h, reached a maximum at 48h and thereafter decreased. The amount of label detected in the γ-glutamyl peptide fractions was below 20% of the total label at any time. It is concluded that in intact plants (at the growth stage used) the γ-glutamyl peptides are not the immediate precursors of the S-alkenyl-L-cysteine sulphoxides. The major S-alkenyl-L-cysteine sulphoxide in onion was found to be compartmentalized mainly within the endoplasmatic reticulum.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00033876
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  • 5
    Electronic Resource
    Electronic Resource
    Production and characterization of intergeneric somatic hybrids through protoplast electrofusion between potato (Solanum tuberosum) and Lycopersicon pennellii (1994)
    Springer
    Plant cell, tissue and organ culture 37 (1994), S. 137-144 
    Details
    ISSN: 1573-5044
    Keywords: electrofusion ; L. pennellii ; protoplasts ; S. tuberosum ; salt tolerance ; somatic hybrids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mesophyll protoplasts of Lycopersicon pennelli Corr., a wild relative of tomato, were electrofused with those from a dihaploid potato clone, cv Nicola, with the objectives of transferring saline tolerance from L. pennellii to cultivated potato. 150 calli were selected from the fusion experiments, finally giving 2 hybrid shoots. Their hybrid nature was verified by examining isoenzyme patterns for esterases (EST), peroxidase (PRX), phosphogluconate dehydrogenase (6-PGD), and glutamate oxaloacetate transaminase (GOT). The hybrid plants had an intermediate morphology, and grew vigorously in vitro. When transplanted to soil, they were less vigorous, due to difficulties in rooting, but were still capable of flowering, and forming short stolons and mishaped tubers, probably resulting from the effects of gene dosage due to the novel association of two genomes from a tuberizing (potato) and a non tuberizing species (L. pennellii). The characteristics of such mishaped tubers provided strong evidence of a hybrid nature for the selected plants. The hybrid plants were highly sterile, producing only 3–7% viable pollen. Tests for salt tolerance showed that the growth of the somatic hybrid plants was reduced by 50% as for L. pennellii, whilst potato did not grow at all under saline conditions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00043607
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  • 6
    Electronic Resource
    Electronic Resource
    Isolation of cells and protoplasts from leaves of in vitro propagated peach (Prunus persica) plants (1994)
    Springer
    Plant cell, tissue and organ culture 36 (1994), S. 99-105 
    Details
    ISSN: 1573-5044
    Keywords: cell division ; peach ; protoplasts ; Prunus persica ; tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Yields of 106–108 peach mesophyll cells and protoplasts · gfw-1 were obtained depending on factors such as digesting enzymes, and leaf size. Onozuka R-10 (2%) in combination with Macerase (0.5%) was found best for protoplast isolation and mediocre for cell isolation among several enzyme combinations tested. Viability was 90% for protoplasts and 60% for cells. Pectolyase Y23 was found to be ineffective in our investigation. Small leaves, 4–10 mm in length, were a superior source for protoplast isolation than medium or big expanded leaves, 22–30 mm in length. The high yields of protoplasts could be obtained only when keeping the ratio of leaf biomass to volume of digesting enzyme solution under 20 mg ml-1. Purification of protoplasts on a sucrose gradient yielded about 107 protoplasts · gfw-1, however, the preparation was still contaminated by intact cells. Protoplasts were cultured under different growth regulators and physical conditions. Limited growth and division of protoplasts embedded in agarose drops were observed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00048320
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  • 7
    Electronic Resource
    Electronic Resource
    Highly efficlent plant regeneration from mesophyll protoplasts of Indian field cultivars of tomato (Lycopersicon esculentum) (1994)
    Springer
    Plant cell, tissue and organ culture 36 (1994), S. 255-258 
    Details
    ISSN: 1573-5044
    Keywords: tomato ; protoplasts ; plant regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: 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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00037728
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  • 8
    Electronic Resource
    Electronic Resource
    Expression of human α-interferon cDNA in transgenic rice plants (1994)
    Springer
    Plant cell, tissue and organ culture 36 (1994), S. 197-204 
    Details
    ISSN: 1573-5044
    Keywords: interferon ; Lipofectin ; protoplasts ; rice ; transgenic plant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The plasmid pIG3031 containing human α-interferon cDNA and the neomycin phosphotransferase II coding sequence was successfully transferred into rice protoplasts (Indica type rice) by Lipofectinmediated transformation. Assays for NPT II enzyme activity indicated that the transformation frequency was 10%. Transgenic plants were regenerated from transformed calli. Southern blots showed that the human α-interferon cDNA sequence was present in rice DNA. RNA slot blots indicated apparent transcription of human α-interferon cDNA under the control of the plant-active promoter PI'. Extracts of transgenic cell cultures and plants contained apparent interferon activity as measured by resistance of a human amniotic cell line to viral infection in the presence of plant extracts. These studies demonstrate that human α-interferon cDNA may be correctly expressed in rice cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00037720
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  • 9
    Electronic Resource
    Electronic Resource
    Isolation and culture of protoplasts from young sporophytes ofSalvinia natans aseptically obtained by co-culture of female and male gametophytes (1994)
    Springer
    Plant cell, tissue and organ culture 36 (1994), S. 237-242 
    Details
    ISSN: 1573-5044
    Keywords: aseptic culture ; female gametophyte ; heterospory ; male gametophyte ; protoplasts ; Salvinia natans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Sporophytes were aseptically obtained by co-culture of female and male gametophytes derived from two types of spores (megaspores and microspores) of the heterosporous fernSalvinia natans All. Protoplasts isolated enzymatically from juvenile leaflets of sporophytes were cultured in a 1/10 Murashige and Skoog's medium containing 2.2 μM naphthalene acetic acid, 2.2 μM 6-benzyl-aminopurine, 0.35 M mannitol, and 0.05 M sucrose. Cell division took place within 6 days of culture, and cell-clusters composed of 9–10 cells were observed after 30 days of culture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00037725
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  • 10
    Electronic Resource
    Electronic Resource
    Single-cell cloning of a crown gall from protoplasts regenerated in hormone-free medium. Establishment of pure transformed cell-lines of Catharanthus roseus (1994)
    Springer
    Plant cell, tissue and organ culture 37 (1994), S. 25-30 
    Details
    ISSN: 1573-5044
    Keywords: Catharanthus roseus ; crown gall tumor ; heterokaryons ; protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts enzymatically isolated from cell line of Catharanthus roseus G. Don crown gall, were cultured at high density (105 P ml-1) in modified B5 liquid medium (Gamborg et al. 1976). In the absence of growth regulators C. roseus protoplasts were able to regenerate a cell-wall, divide and, subsequently, yield very numerous clones in the absence of growth regulators. After two weeks, the cultures were greatly diluted in order to obtain clones of single-cell origin. Most of the clones individually transferred onto solid medium can proliferate indefinitely, without growth regulators. Among analyzed clones, 90% were nopaline positive. Their ajmalicine and serpentine content was compared with that of the parental crown gall line, and was found to be low. The CR10 protoplasts were very easy to grow, they were an interesting model for the development of pure tumorous lines. Moreover, we found that the tumorous protoplasts were useful for cell fusion experiments or for the delicate culture of tree protoplasts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00048113
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  • 11
    Electronic Resource
    Electronic Resource
    Recovery of transgenic trees after electroporation of poplar protoplasts (1994)
    Springer
    Transgenic research 3 (1994), S. 13-19 
    Details
    ISSN: 1573-9368
    Keywords: gene transfer ; electroporation ; stable transformation ; protoplasts ; transgenic trees ; Populus tremula x P. alba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts from leaflets ofin vitro cuttings were electroporated in osmotically adjusted and buffered solutions containing plasmid DNA: pABD1, carrying thenptII gene for resistance to neomycin; pGH1, carrying a mutant acetolactate synthase gene,als, for resistance to sulfonylurea; and pGSFR781A, carrying a synthetic phosphinothricin acetyltransferase (pat) for resistance to phosphinothricin (Basta). Gene transfer was repeatedly efficient, without use of carrier DNA, in the range of one transformant for 105 to 104 protoplast-derived cell colonies. This was probably due to the high plating efficiency (30%) of protoplasts in our culture process. Selection for expression of foreign genes was applied in liquid medium and repeatedly achieved with 30 μM paromomycin for NPTII, 200 nM chlorsulfuron for the mutant ALS ofArabidopsis and 25 μM phosphinothricin for PAT expression. Integration of foreign genes into genomic DNA of resistant poplar trees was demonstrated by Southern blot hybridizations, which revealed that for some transformants practically no other part of the vector plasmid than the selected gene was integrated. Effective processes for protoplast culture, efficient selection at the cell colony stage and gene transfer will provide new possibilities in poplar breeding.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01976022
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