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  • 1
    Electronic Resource
    Electronic Resource
    Chloroplast transfer in Nicotiana based on metabolic complementation between irradiated and iodoacetate treated protoplasts (1981)
    Springer
    Planta 152 (1981), S. 341-345 
    Details
    ISSN: 1432-2048
    Keywords: Chloroplast transfer ; Nicotiana ; Protoplasts fusion and inactivation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts of a light sensitive plastome mutant of Nicotiana tabacum (2 n=48) were irradiated and fused with iodoacetate-treated Nicotiana plumbaginifolia (2 n=20) protoplasts. Treated parental protoplasts were unable to divide. Metabolic complementation, however, helped the recovery of interspecific fusion products which survived and formed calli. Altogether 40 clones were investigated. N. plumbaginifolia plants were obtained in 15 clones (38%), somatic hybrids in 23 clones, and both types of regenerates were found in 2 clones. Irradiation therefore significantly increased the frequency of segregant formation with the non-irradiated N. plumbaginifolia nuclei (the frequency was 1.4% in the absence of irradiation). Regenerated plants in most cases (31 out of 34) contained chloroplasts from the irradiated parent. In 6 clones plants were obtained with both types of chloroplast. Thus, irradiated N. tabacum chloroplasts had an improved chance of dominating the heterokaryonderived cells, many of which contained N. plumbaginifolia nucleus. The system described should be generally applicable for the transfer of chloroplasts without the use of selectable genetic markers.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00388259
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  • 2
    Electronic Resource
    Electronic Resource
    Improved expression of streptomycin resistance in plants due to a deletion in the streptomycin phosphotransferase coding sequence (1988)
    Springer
    Molecular genetics and genomics 214 (1988), S. 456-459 
    Details
    ISSN: 1617-4623
    Keywords: Chimeric gene ; Mutant streptomycin phosphotransferase ; Non-lethal screen ; Streptomycin resistance ; Transgenic Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Previous studies have shown that a chimeric streptomycin phosphotransferase (SPT) gene can function as a dominant marker for plant cell transformation. The SPT marker previously described by Jones and co-workers has a limited value since it conferred a useful level of resistance only to a fraction (10%) of Nicotiana plumbaginifolia transgenic lines. Expression of resistance was species specific: no such resistant transformants were found in N. tabacum. In this paper we describe an improved SPT construct that utilizes a mutant Tn5 SPT gene. The mutant gene, SPT *, encodes a protein with a two amino acid deletion close to its COOH-terminus. In N. tabacum cell culture the efficiency of transformation with the improved streptomycin resistance marker was comparable to kanamycin resistance. When the chimeric SPT * gene was introduced linked to a kanamycin resistance gene, streptomycin resistance was expressed in most of the transgenic N. tabacum lines.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00330480
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  • 3
    Electronic Resource
    Electronic Resource
    Streptomycin and lincomycin resistances are selective plastid markers in cultured Nicotiana cells (1990)
    Springer
    Molecular genetics and genomics 221 (1990), S. 245-250 
    Details
    ISSN: 1617-4623
    Keywords: Lincomycin resistance ; Nicotiana ; Plastid markers ; Selection ; Streptomycin resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Resistance to streptomycin and lincomycin in plant cell culture is used as a color marker: resistant cells are green whereas sensitive cells are white on the selective medium. Streptomycin and lincomycin at appropriate concentrations do not kill sensitive Nicotiana cells. The selective value of plastid ribosomal DNA mutations, conferring resistance to streptomycin and lincomycin, was investigated by growing heteroplastidic cells on a selective medium. The heteroplastidic cells were obtained by protoplast fusion, and contained a mixed population of streptomycin resistant plastids from the N. tabacum line Nt-SR1-Kan2, and lincomycin resistant plastids from the N. plumbaginifolia line Np-LR400-Hyg1. Clones derived from protoplast fusion were selected by kanamycin and hygromycin resistance, transgenic nuclear markers. Somatic hybrids were then grown on a selective streptomycin or lincomycin medium, or in the absence of either drug to a 50 to 100 mg size callus. Southern analysis of a polymorphic region of plastid DNA (ptDNA) revealed that somatic hybrids grown on streptomycin contained almost exclusively ptDNA from the streptomycin resistant parent, somatic hybrids grown on lincomycin contained almost exclusively ptDNA from the lincomycin resistant parent whereas somatic hybrids grown in the absence of either drug contained mixed parental plastids. Sensitive ptDNA was below detection level in most clones on selective medium, but could be recovered upon subsequent culture in the presence of the appropriate drug. The drugs streptomycin and lincomycin provide a powerful selection pressure that should facilitate recovery of plastid transformants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00261727
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  • 4
    Electronic Resource
    Electronic Resource
    Mutation proximal to the tRNA binding region of the Nicotiana plastid 16S rRNA confers resistance to spectinomycin (1991)
    Springer
    Molecular genetics and genomics 228 (1991), S. 316-319 
    Details
    ISSN: 1617-4623
    Keywords: Nicotiana tabacum ; Plastid mutants ; 16S ribosomal RNA ; Spectinomycin resistance ; Streptomycin resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Nicotiana tabacum lines carrying maternally inherited resistance to spectinomycin were obtained by selection for green callus in cultures bleached by spectinomycin. Two levels of resistance was found. SPC1 and SPC2 seedlings are resistant to high levels (500 μg/ml), SPC23 seedlings are resistant to low levels (50 μg/ml) of spectinomycin. Lines SPC2 and SPC23 are derivatives of the SR1 streptomycin-resistant plastome mutant. Spectinomycin resistance is due to mutations in the plastid 16S ribosomal RNA: SPC1, an A to C change at position 1138; SPC2, a C to U change at position 1139; SPC23, a G to A change at position 1333. Mutations similar to those in the SPC1 and SPC2 lines have been previously described, and disrupt a conserved 16S ribosomal RNA stem structure. The mutation in the SPC23 line is the first reported case of a mutation close to the region of the 16S rRNA involved in the formation of the initiation complex. The new mutants provide markers for selecting plastid transformants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00282483
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  • 5
    Electronic Resource
    Electronic Resource
    A dominant nuclear streptomycin resistance marker for plant cell transformation (1987)
    Springer
    Molecular genetics and genomics 210 (1987), S. 86-91 
    Details
    ISSN: 1617-4623
    Keywords: Chimeric gene ; Nicotiana ; Streptomycin phosphotransferase ; Streptomycin resistance ; Transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Plant cells in photoheterotrophic culture respond to streptomycin by bleaching and retarded growth but no cell death. A new genetic marker for plant cell transformation has been developed that is based on the expression of the enzyme streptomycin phosphotransferase (SPT), and confers the ability to form green colonies on a selective medium. Coding sequences of SPT from the bacterial transposon Tn5 were placed under the control of gene expression signals derived from the Agrobacterium Ti plasmid Ach5. The 5′ end of the SPT gene has been replaced with the promoter region of the gene coding for the first enzyme of agropine biosynthesis, the 3′ end with that of the enzyme octopine synthase. The chimeric SPT gene has been linked to a selectable kanamycin resistance gene, and introduced into Nicotiana tabacum and Nicotiana plumbaginifolia by selection for the linked kanamycin resistance marker. Streptomycin resistance was expressed in some but not all of the kanamycin-resistant lines and was transmitted to the seed progeny as a dominant nuclear trait.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00337762
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