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  • 1
    ISSN: 1432-2048
    Keywords: Daucus ; Embryogenic potential ; Gene expression ; mRNA ; Somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Somatic embryogenesis can be synchronized by enriching carrot (Daucus carota L.) suspension cultures for small, dense clusters of cells termed proembryogenic masses (PEMs). Gene-expression programs of PEMs were compared with those of embryonic and mature tissues by in-vitro translation of representative mRNA populations and by nucleic-acid hybridization. Analysis of invitro-translated polypeptides by two-dimensional polyacrylamide gel electrophoresis revealed striking similarities between the mRNA populations of PEM and torpedo-stage embryos; substantial differences, however, were observed when in-vitro translation products of PEMs and torpedo embryos were compared with those of hypocotyls and leaves. Northern blots of RNA isolated from PEMs, staged embryos, and mature carrot tissues were hybridized with cDNA probes for Dc3, Dc5 and Dc13; these cDNA recombinants represent mRNAs that are regulated during carrot somatic embryogenesis. The pattern of expression of these embryo-regulated transcripts was similar in PEMs and somatic embryos but differed in other carrot tissues. These results indicate that many of the molecular processes of embryogenesis are already established in PEMs in the presence of auxin. Additional experiments indicate the utility of Dc3 as a molecular marker for the acquisition of embryogenic potential.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2048
    Keywords: Cell tracking ; Daucus ; Development (single cell) ; 2,4-Dichlorophenoxyacetic acid ; Phytagel (cell immobilisation) ; Somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cell-tracking system was established to determine the capability of individual single suspension cells of carrot (Daucus carota L.) to develop into somatic embryos. When immobilised in phytagel, 127 out of 30 318 single suspension cells smaller than 22 μm in diameter developed into a somatic embryo. Single cells present at the start of the experiment were classified on the basis of their morphology into five groups: small spherical vacuolated cells; small spherical cytoplasm-rich cells; oval vacuolated cells; elongated vacuolated cells and cells that could not be classified into either one of these groups. Single cells of all morphologically distinguishable single cell types developed into somatic embryos with a frequency that varied between 19 and 100 somatic embryos per 10 000 cells. This suggests that the capability of individual single cells to form somatic embryos is not restricted to a particular cell type distinguishable on the basis of its morphology. Three major pathways were observed during development. Oval and elongated cells developed into somatic embryos via an asymmetrical cell cluster. Spherical cells developed via a symmetrical cell cluster into somatic embryos. Before formation of a somatic embryo, cells of a more variable initial morphology first developed aberrantly shaped cell clusters. This suggests that the developmental pathway leading to a somatic embryo can be predicted by the initial single-cell morphology.
    Type of Medium: Electronic Resource
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