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  • 1
    Electronic Resource
    Electronic Resource
    Using firefly luciferase to identify the transition from transient to stable expression in bombarded wheat scutellar tissue (1998)
    Copenhagen : Munksgaard International Publishers
    Physiologia plantarum 102 (1998), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Transformation of wheat by the biolistic procedure is unpredictable and inefficient. To gain insight into the fate of transgenes introduced into wheat scutellar tissue by particle bombardment, the expression of the firefly luciferase gene was followed using low-light video imaging. Luciferase expression can be detected as early as 40 to 50 min after bombardment. After 48 h, the levels of transient expression in individual scutella fall rapidly, becoming undetectable 10 to 20 days later. Luciferase activity was observed to recover in a small percentage of the material and it was from this material that transformed plants, stably expressing luciferase, were recovered. We concluded that the calluses which recover luciferase expression are stably transformed and we have termed the transition period between transient and stable expression the transformation boundary. The percentage of scutella displaying luciferase activity, as measured at 30 days post-bombardment, was judged a realistic measure of the efficiency of the transformation procedure. The results of our experiments suggest that the selection and regeneration of plants were not major factors contributing to the poor transformation efficiencies associated with biolistic transformation. The results demonstrate that luciferase can be used to assess rapidly and quantify the efficiency of the transformation procedure without the need to produce transformed plants. This will allow different procedures to be rapidly assessed and compared and should provide valuable insight into the conditions required to improve the efficiency of DNA integration and stable expression in species recalcitrant to transformation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1034/j.1399-3054.1998.1020313.x
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  • 2
    Electronic Resource
    Electronic Resource
    Transfer of methomyl and HmT-toxin sensitivity from T-cytoplasm maize to tobacco (1991)
    Springer
    Molecular genetics and genomics 229 (1991), S. 405-412 
    Details
    ISSN: 1617-4623
    Keywords: T-URF13 ; Tobacco ; Methomyl ; Transformation ; Maize
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The mitochondrial gene, T-urf13, which is unique to the T-cytoplasm of maize, has been expressed in tobacco plants using the Cauliflower Mosaic Virus 35S promoter. Tobacco plants expressing T-urf13 exhibit a variety of responses to methomyl. Leaf discs and petiole sections bleach when exposed to methomyl or HmT-toxin; this effect increases with the age of the tissue. The bleaching effect is not however observed when light is excluded. Plants homozygous for T-urf13 exhibit extreme sensitivity when sprayed with methomyl. The growth of seedling which are either homozygous or heterozygous for T-urf13 is inhibited by methomyl and by kanamycin, whereas seedlings from untransformed tobacco or tobacco which has lost the T-urf13 gene through segregation are sensitive to kanamycin but develop normally when exposed to methomyl. The results demonstrate that T-URF13 need not be specifically targeted to the mitochondrion for it to induce methomyl or HmT-toxin sensitivity in tobacco.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00267463
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  • 3
    Electronic Resource
    Electronic Resource
    Isolation and characterization of the wheat prolyl isomerase FK506-binding protein (FKBP) 73 promoter (2000)
    Springer
    Plant molecular biology 42 (2000), S. 489-497 
    Details
    ISSN: 1573-5028
    Keywords: ABA-responsive ; FKBP73 ; promoter ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The wheat FK506-binding protein (FKBP) 73 is a member of the peptidyl prolyl cis-trans isomerase gene family, which catalyses the interconversion between the cis and trans forms of the peptide bond preceding proline residues in proteins. A 3.5 kb sequence 5′ upstream of the ATG codon of the wheat FKBP73 was isolated from a wheat genomic library, and characterized by deletion analysis and transient expression in wheat embryos. The 1517 bp fragment is referred to as the full promoter due to the maximal activity of the fused luciferase reporter gene. Sequence analysis revealed the presence of three abscisic acid (ABA)-responsive elements (ABREs) proximal to coupling elements (CE1-like), a putative lectin box, two putative binding sites for the myb transcription factor and a 36 bp fragment which exhibits 100% identity to the pSau3A9 clone located in the centromeric region of wheat chromosomes. In a transient expression assay the promoter preserved the tissue specificity described in vivo, namely it is expressed only in germinating embryos and young shoots. The promoter was induced 1.9-fold by ABA, the minimal promoter was designated at −221 and the TATA box located at −137. The inducibility by ABA and the expression during germination may indicate that FKBP73 belongs to the group of genes induced by ABA upon germination.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006345411056
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