Optimization of Chitinase gene transfer to tomatoes of Bonab cultivar using Agrobacterium tumefaciens

Tomato is an important economic crop with nutritional and scientific value for study. Due to its agronomic importance and the sensitivity to biotic and abiotic stresses, using new methods in breeding this crop is necessary. In this study, the effects of some factors, such as pre-cultivation time (1, 2, 3 and 4 days), Agrobacterium concentration (OD600nm = 0.4, 0.6, 0.8, and 1), inoculation time (5, 10, 15 and 20 minutes), Acetosyringone concentration (0, 100, 150, and 200 μM), co-cultivation period (1, 2, 3 and 4 days) and different concentrations of the kanamycin (5, 10, 15 and 20 mg/l) on the transformation of a local cultivar of tomato called Bonab, were evaluated. The maximum transformation efficiency was obtained at the OD600= 0.6. Also, the highest transgenic plants were obtained in the 10 minutes of inoculation and 2 days of co-cultivation. The presence of Acetosyringone had an increasing effect on the tomato transformation, and the highest percentage of transformation was obtained at 150 M of Acetosyringone. Finally, the presence of a Chitinase gene in the probable transgenic plants was confirmed by PCR. The optimal conditions that were obtained in this study can be used to transfer target genes to this tomato cultivar.