High efficiency Agrobacterium rhizogenes-mediated transformation of Saponaria vaccaria L. (Caryophyllaceae) using fluorescence selection

• Published in: Plant cell reports Vol. 26; no. 9; pp. 1547 - 1554
• Main Authors: Schmidt, Janice F, Moore, Maria D, Pelcher, Lawrence E, Covello, Patrick S
• Format: Journal Article
• Language: English
• Published: Berlin Berlin/Heidelberg : Springer-Verlag 01-09-2007; Springer; Springer Nature B.V
• Subjects: Agrobacterium rhizogenes; Biological and medical sciences; Biotechnology; Caryophyllaceae; Caryophyllaceae - genetics; Cultivars; Drug Resistance, Microbial; Eukaryotic cell cultures; Fluorescence; Fundamental and applied biological sciences. Psychology; Genetic Techniques; Green fluorescent protein (GFP); Green Fluorescent Proteins - metabolism; Methods. Procedures. Technologies; Miscellaneous; Plant cells and fungal cells; Plant Roots - growth & development; Plants, Genetically Modified; Polymerase Chain Reaction; Proteins; Rhizobium - physiology; Rhizobium rhizogenes; Roots; Saponaria; Selection, Genetic; Spectrometry, Fluorescence; Transformation; Transformation, Genetic; Vaccaria; Vaccaria hispanica; Hairy root; Transformation; Genetic transformation; Vaccaria hispanica; Selection; Tissue culture; Fluorescence; Caryophyllaceae; Dicotyledones; Angiospermae; Green fluorescent protein; Green fluorescent protein (GFP); Saponaria vaccaria; Bacteria; Adventitious root; Spermatophyta; Rhizobiaceae; Agrobacterium rhizogenes
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s00299-007-0369-9

A highly efficient and convenient method for the Agrobacterium rhizogenes-dependent production of transformed roots of Saponaria vaccaria L. (Caryophyllaceae) is described. The parameters tested and optimized include S. vaccaria cultivar, explant type, Agrobacterium rhizogenes strain and culture conditions. For cotransformation using additional recombinant T-DNA-containing A. rhizogenes strains, use of neomycin phosphotransferase and enhanced green fluorescent protein genes as selectable markers were tested alone and in combination. Optimal results, yielding a minimum of one transformed root per explant, were obtained using the cultivar Pink Beauty, the A. rhizogenes strain LBA9402 and internode explants precultured on a phytohormone mixture. Selection of cotransformed roots by observation of enhanced green fluorescent protein fluorescence alone was highly effective and convenient.