2-deoxyglucose selection system for Agrobacterium-mediated transformation of Solanum lycopersicon and Nicotiana tabacum plants

Abstract

An alternative marker to antibiotic and herbicide genes, 2-deoxyglucose-6-phosphate phosphatase (DOGR1) gene was used to produce transgenic tomato (Solanum lycopersicon) and tobacco plants (Nicotiana tabacum) via Agrobacterium-mediated transformation approach. A binary vector, pBIDOG with T-DNA cassette consists of CaMV35S-DOGR1-nos was mobilized into Agrobacterium tumefaciens LBA4404 via electroporation and subsequently transformed into tomato and tobacco tissues. Selection of transformants of both plants was carried out by using 600 mg/l 2-deoxyglucose (2-DOG) as selection agent. 2-DOG resistant calli were regenerated into shoots and PCR analysis revealed that 75% and 71% of putative transgenic tomato and tobacco shoots were positive for DOGR1 gene, respectively. Southern blot analysis was carried out to confirm the integration of transgene. The expected ~1.5 kb band was obtained to prove the integration of DOGR1 gene into the plant genome. Transgenic tomato and tobacco plants were successfully regenerated without any phenotypic changes when compared to wild-type plants. Therefore, this alternative selection system can be applied to other plant transformation systems to produce transgenic plants free from antibiotic and herbicide genes, especially for important economic crops. More importantly, this selection system does not lead to any extent hazard to human and animal that subsequently will improve public acceptance of future genetically modified products.