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  • 1
    Electronic Resource
    Electronic Resource
    Isolation of an isoleucine-valine-requiring auxotroph fromDatura innoxia cell cultures by arsenate counterselection (1983)
    Springer
    Planta 159 (1983), S. 12-17 
    Details
    ISSN: 1432-2048
    Keywords: Cell culture (auxotroph isolation) ; Datura ; Isoleucine ; Mutant counterselection ; Valine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cells of adenine(Ad1)- and pantothenate(Pn1)-requiring cells lines ofDatura innoxia Mill. were killed by 2 mM arsenate in the medium when growing but were not killed when in a non-growing state which was induced by starvation for required metabolites. This observation confirmed that arsenate might be useful as a counterselection agent to isolate new auxotrophs from plant cell cultures. Arsenate counterselection using predominantly haploidDatura cell suspensions led to the isolation of a cell line (I–IV) for which isoleucine and valine were both necessary and sufficient for growth. The cell line has been maintained in solid and liquid culture for about three years. The arsenate counterselection technique devised requires improvement. In some experiments, many colonies survived arsenate treatment; in others, there were no survivors. In the former experiments, putative but unstable auxotrophs were regularly observed but all were shown not to be auxotrophic after several retests.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00998808
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  • 2
    Electronic Resource
    Electronic Resource
    The isolation of auxotrophs from Datura innoxia Mill. cell cultures following recovery of arsenate-treated cells on feeder plates (1984)
    Springer
    Planta 160 (1984), S. 168-173 
    Details
    ISSN: 1432-2048
    Keywords: Cell culture ; Datura ; Mutant (counterselection) ; Nitrate reductase (deficient mutant) ; Threonine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Wild-type (Ph1) and adenine-requiring (Ad1) cell lines of Datura innoxia Mill. were used in experiments to evaluate arsenate as a growth-lethal compound and its use as a counterselection agent. These experiments led to the devising of methods for the recovery of Ph1 and Ad1 cells after arsenate treatment when plated at low density on feeder plates. The modified counterselection technique was then used to isolate three new auxotrophs from mutagenized suspensions of Ph1 cells, two of which were partially characterized. One, C18, requires casein hydrolysate for growth and lacks an active nitrate reductase; the other, JM3, will grow only when the medium contains threonine.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00392866
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  • 3
    Electronic Resource
    Electronic Resource
    Transplantation of isolated nuclei into plant protoplasts (1986)
    Springer
    Planta 168 (1986), S. 29-35 
    Details
    ISSN: 1432-2048
    Keywords: Auxotrophy ; Datura ; DNA transfer ; Nuclear transplantation ; Protoplast ; Prototrophy ; Vicia (nuclear transplantation)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The uptake of isolated nuclei from Vicia hajastana Grossh. cells into protoplasts of an auxotrophic cell line of Datura innoxia P. Mill. was induced under the influence of polyethylene glycol and Ca2+ at pH 6.8. The frequency of nuclear uptake varied from 0.8 to 2.3% and that of the recovery of prototrophic clones from 10-5 to 6·10-4. The prototrophic nuclear fusion products following nuclear uptake could be rescued by initial culture of the protoplasts in non-selective conditions and by the subsequent use of feeder cell layers to support the growth of surviving colonies on a selective medium. The presence of Vicia genomic DNA in some prototrophic clones was confirmed by dot-blot hybridization using Datura and Vicia DNA probes. In certain transformed clones, the recovery of prototrophy was accompanied by the restoration of morphogenetic potential. Welldeveloped shoots typical of wild-type Datura could be regenerated employing an appropriate regeneration medium.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00407005
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  • 4
    Electronic Resource
    Electronic Resource
    A transient duplication of the acetolactate synthase gene in a cell culture of Datura innoxia (1987)
    Springer
    Theoretical and applied genetics 74 (1987), S. 417-422 
    Details
    ISSN: 1432-2242
    Keywords: Gene amplification ; Datura ; Sulfonylurea resistance ; Plant cell culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 2.0 kb fragment of the yeast ILV2 gene, which codes for the target enzyme acetolactate synthase (ALS) of the herbicide chlorsulfuron, was shown to hybridize to the nuclear DNA of a haploid cell culture of Datura innoxia P. Mill. Nuclear DNA of a chlorsulfuron resistant line of D. innoxia, CSR6, gave a prominent 2.65 kb band when cleaved by either EcoRI or HindIII. The 2.65 kb band has been shown to hybridize with the yeast ILV2 probe. A herbicide resistant line descended from CSR6 by continuous culture resulted in the loss of the 2.65 kb restriction fragment. These observations suggest that CSR6 resulted from a large tandem duplication of the ALS gene and that a point mutation for herbicide resistance in an ALS gene repeat unit of the duplication was selected during subsequent growth of the resistant line.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00289814
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  • 5
    Electronic Resource
    Electronic Resource
    Partial characterization of two stable auxotrophic cell strains of Datura innoxia Mill (1980)
    Springer
    Planta 149 (1980), S. 480-484 
    Details
    ISSN: 1432-2048
    Keywords: Adenine ; Auxotroph ; Cell cultures, variant lines ; Datura ; Pantothenate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A growth analysis of several presumptive “leaky” auxotrophs from Datura innoxia suspension cultures led to the discovery of an adenine-requiring cultures led to the discovery of an adenine-requiring cell strain (Ad1). Both Ad1 and Pn1, a pantothenate-requiring strain isolated earlier from these cultures, still require either adenine or pantothenate for growth after more than one year in culture. Attempts to select prototrophic revertants have failed. Ad 1 also grew well in a medium containing either 5-aminoimidazole-4-carboxamide ribotide or inosine instead of adenine; Pn 1 with pantoic acid alone but not at all in the presence of β-alanine or α-ketoisovalerate alone instead of pantothenate. Pn 1 cells starved of pantothenate for up to 4d and Ad1 of adenine for 10d or more resumed growth when transferred to appropriately supplemented media. Wild-type Datura cells grown on unsupplemented medium would not crossfeed the required nutrients to the auxotrophs. The starvation and cross-feeding experiment showed that both auxotrophs could be used in reconstruction experiments to develop enrichment-selection techniques for the isolation of more auxotrophs.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00385752
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