High efficiency Agrobacterium rhizogenes-mediated transformation of Saponaria vaccaria L. (Caryophyllaceae) using fluorescence selection.
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.
Schmidt
Janice F
JF
Plant Biotechnology Institute, 110 Gymnasium Place, S7N OW9 Saskatoon, SK, Canada.
Moore
Maria D
MD
Pelcher
Lawrence E
LE
Covello
Patrick S
PS
D029748:Caryophyllaceae / Q000235:genetics*; D004352:Drug Resistance, Microbial; D005453:Fluorescence; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018517:Plant Roots / Q000254:growth & development; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D012231:Rhizobium / Q000502:physiology*; D012641:Selection, Genetic; D013050:Spectrometry, Fluorescence; D014170:Transformation, Genetic
D049452:Green Fluorescent Proteins