Optimization of Agrobacterium Tumefaciens Mediated Gene Transfer Method in Tomato (Solanum Lycopersicum L.)

Abstract

Tomato (Solanum lycopersicum L.) is one of the most important vegetable crops of genus solanum which belongs to Solanaceae family. It is very versatile and is grown either for fresh fruits or for processing. Worldwide, 170.8 million tons of tomatoes were produced in the world. At present, India contributes 11 % of world tomato production and it is grown in an area of 7,97,000 hectare with productivity of 20.70 MT/hectare. In present study, Cotyledon explants from 10 day old in-vitro seedling were used for callus regeneration and optimization of transformation protocol. Different concentrations of auxins and cytokinins combinations had a distinct effect on callus induction from explants. Best 85% callus induction frequency was obtained when MS medium was supplemented with 3 mg/l 2, 4-D and 0.25 mg/l Kinetin. The transformation was carried out using the Agrobacterium stain EHA 105, containing the binary vector pCAMBIA 1301 harboring GUS as reporter gene. The infection time of 20 minutes for callus & cotyledonary leaf explants with 2 days of co-cultivation period was found 13.3% suitable for the maximum transformation frequency. The transient GUS gene was confirmed by X-Gluc solution which showed blue color formation through GUS assay.