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  • 1
    ISSN: 1572-9788
    Keywords: Alstroemeria ; bar ; GUS ; luciferase ; ubiquitin promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Transgenic plants were obtained after particle bombardment of embryogenic callus derived from stem segments of two tetraploid Alstroemeria genotypes with plasmids containing different selection/reporter genes. Firstly, a plasmid containing a firefly luciferase reporter gene driven by the maize ubiquitin promoter (Ubi1), was bombarded into both friable embryogenic callus and proembryos. Transient and stable expression of luciferase was visually detected by a luminometer. This selection method is non-destructive and can be applied over the whole developmental process from callus to embryo and plantlet. Molecular proof of transformation was obtained both by PCR analysis and Southern hybridization. Secondly, a plasmid containing the bar gene together with an uidA gene coding for β-glucuronidase both driven by the Ubi1 promoter was bombarded into proembryos. The transgenic callus was effectively selected from the callus clumps four months after bombardment on a medium containing 5 mg/l phosphinotricin (PPT). Selection by PPT was efficient and labour-saving. Stable expression of GUS was confirmed by the histochemical staining assay and molecular proof was obtained by PCR analysis.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 52 (1998), S. 165-169 
    ISSN: 1573-5044
    Keywords: age ; in vitro ; Inca lily ; monocots ; micropropagation ; regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Direct shoot regeneration was induced from leaf explants of Alstroemeria. The explants contained a leaf blade and a small portion of stem node, which were cut from the erect shoots of in vitro multiplicated plantlets. The shoot regeneration capacity of the excised leaf explants was significantly related to the position of the explant on the stem. The youngest explant which was located closest to the shoot apex gave the highest response. A gradient response toward the shoot apex was observed in percentage of shoot regeneration and in the number of shoots per regenerating explant. Histological studies revealed that the shoots were initiated at the leaf axils. The origin of the adventitious buds was located at the epidermal layer of stem peripheral cells.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-5060
    Keywords: line ; onion ; regeneration ; shallot ; suspension culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Suspension cultures were initiated from callus cultures of Allium cepa, which had been precultured on a solidified medium for 7 months. For another 3 months the 83 callus lines were kept in suspension culture. Each line is derived from a single zygotic embryo, 24 lines from onion and 59 lines from shallot. Of these, 20 suspension lines showed adequate growth and were used to test the effect of cytokinins on plant regeneration. On average, 1.25% of the calli produced shoots on a growth regulator-free medium (MS30: MS basal medium supplemented with 30 g/l sucrose). Thus plant regeneration decreased significantly with time because the overall average plant regeneration frequency was 35.5% after 3 months of culture on solidified medium. Plant regeneration after 3 months proved not to be a reliable predictor of plant regeneration after 10 months in vitro culture. Overall only 0.25–0.88% of the calli produced shoots when the regeneration medium was supplemented with different types and concentrations of cytokinins. Contrary to expectations, the type and concentration of cytokinins could not increase the shoot regeneration capacity. However as expected, the increasing cytokinin concentration (especially TDZ at the range of 1–5 mg/l) led to a decrease in root formation. Plant regeneration proved to be highly dependent upon the line used. The best line was atm24 with an overall shoot regeneration capacity of 4.62% (among the 13 treatments); it had its highest shoot regeneration on MS30 treatment with 12.5%. The results obtained show that for the development of a reliable transformation protocol only young callus material (〈 3 months), which has still a high regeneration potential, can be used.
    Type of Medium: Electronic Resource
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