Evidence that direct DNA uptake through cut shoots leads to genetic transformation of Solanum aviculare Forst.
The reporter genes GUS, NPTII and BAR, either separately or in combination, have been exploited to determine if DNA which can directly enter plants, circulate within the plant and enter nuclei, can also integrate into the genome in a manner which will permit gene expression. Feeding of either seed-derived or adventitious cut shoots of Solanum aviculare with the GUS gene followed by rooting of the shoots and growing on, resulted in all tissues of the plant showing GUS activity as detected cytochemically. Southern blot analysis of plants derived from the adventitious shoots confirmed the presence of the reporter gene in roots. Reporter gene expression was observed also in the F1 generation. If GUS and NPTII or GUS, NPTII and BAR were fed together, then in each case it was possible to have both expression and Southern blot confirmation of each of the genes. There was a relatively high rate of transformation of approximately 5% of the fed stems across all experiments conducted during the present study.
Gahan
P B
PB
School of Biomedical Sciences, King's College London, Guy's Campus, London Bridge, London, UK. pgahan@aol.com
Wyndaele
R
R
Mantell
S
S
Boggetti
B
B
D000900:Anti-Bacterial Agents / Q000378:metabolism; D001692:Biological Transport; D004247:DNA / Q000008:administration & dosage / Q000378:metabolism*; D004351:Drug Resistance; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics; D007612:Kanamycin / Q000378:metabolism; D019868:Kanamycin Kinase / Q000235:genetics; D018520:Plant Shoots / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D012639:Seeds / Q000378:metabolism; D032322:Solanum / Q000235:genetics* / Q000502:physiology*; D014170:Transformation, Genetic
D000900:Anti-Bacterial Agents; D007612:Kanamycin; D004247:DNA; D019868:Kanamycin Kinase; D005966:Glucuronidase