Agrobacterium tumefaciens-mediated transformation of eggplant (Solanum melongena L.) using root explants.
An efficient variety-independent method for producing transgenic eggplant (Solanum melongena L.) via Agrobacterium tumefaciens-mediated genetic transformation was developed. Root explants were transformed by co-cultivation with Agrobacterium tumefaciens strain LBA4404 harbouring a binary vector pBAL2 carrying the reporter gene beta-glucuronidase intron (GUS-INT) and the marker gene neomycin phosphotransferase (NPTII). Transgenic calli were induced in media containing 0.1 mg l(-1) thidiazuron (TDZ), 3.0 mg l(-1) N(6)-benzylaminopurine, 100 mg l(-1) kanamycin and 500 mg l(-1) cefotaxime. The putative transgenic shoot buds elongated on basal selection medium and rooted efficiently on Soilrite irrigated with water containing 100 mg l(-1) kanamycin sulphate. Transgenic plants were raised in pots and seeds subsequently collected from mature fruits. Histochemical GUS assay and polymerase chain reaction analysis of field-established transgenic plants and their offsprings confirmed the presence of the GUS and NPTII genes, respectively. Integration of T-DNA into the genome of putative transgenics was further confirmed by Southern blot analysis. Progeny analysis of these plants showed a pattern of classical Mendelian inheritance for both the NPTII and GUS genes.
Franklin
G
G
Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore 560 012, India. gfrankl2@utnet.utoledo.edu
Lakshmi Sita
G
G
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D046508:Culture Techniques; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000196:embryology / Q000235:genetics / Q000502:physiology*; D018520:Plant Shoots / Q000196:embryology / Q000502:physiology; D030821:Plants, Genetically Modified / Q000196:embryology / Q000235:genetics* / Q000502:physiology; D012038:Regeneration; D012639:Seeds / Q000196:embryology / Q000235:genetics / Q000502:physiology; D032323:Solanum melongena / Q000196:embryology / Q000235:genetics* / Q000502:physiology; D014170:Transformation, Genetic
D005966:Glucuronidase