Responses of Nicotiana tabacum L. Wild-Type to Selection and Elimination Antibiotic Concentrations During Genetic Transformations
Abstract
Agrobacterium-mediated plant transformation requires antibiotics to screen transgenic cells and eradicate residual bacteria after co-cultivation. Both treatments induced phytotoxic effects that impaired the regeneration of the explants. This study determined the optimal minimum concentrations of antibiotics for the selection and elimination of wild-type Nicotiana tabacum L. (West Java accession) using the pBI121 vector containing the kanamycin-resistance gene (nptII). Kanamycin antibiotic sensitivity testing identified 100 ppm as the minimum effective selection concentration, which induced necrosis in all non-transformant explants. A comparative assessment of two broad-spectrum β-lactam antibiotics, augmentin and cefotaxime, was conducted based on three parameters: bacterial contamination rate, shoot height, and root induction percentage. The results indicated that 300 ppm of augmentin demonstrated the best performance. The results demonstrated a 0% contamination rate, accompanied by notably lower phytotoxicity than cefotaxime. These findings establish an optimized antibiotic protocol for efficient N. tabacum transformation with minimal tissue damage.
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DOI: https://doi.org/10.17509/jobs.v3i1.100651
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