Punctured-hypocotyl Method of Agrobacterium-mediated Transformation

Tomato productivity has always been constrained because of abiotic stresses. Transgenic tomatoes are presently being developed to minimize these losses due to abiotic stresses. Agrobacterium-mediated transformation is the most common approach to producing transgenic tomato. However, the effectiveness of the present methods were limited to only a few tomato cultivars. Hence, we still need an appropriate, simple and general procedure effective across all cultivars. Wounding methods, such as puncturing with a syringe needle, may just be the answer.
Using Indian tomato hypocotyl explants, the efficiencies of the punctured-hypocotyl method as well as normal immersion method of Agrobacterium-mediated transformation were compared. All factors influencing transformation efficiency, such as Agrobacterium density and co-cultivation time, were optimized. The transgene integration of the tomato genome was confirmed by PCR and Southern hybridization. Transformation efficiency was found to be greater with the punctured-hypocotyl method compared to the normal immersion method.
This newly developed method is simple, efficient and could be used to transfer important agronomic genes into the tomato genome for the potential improvement in terms of quality and quantity.
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