In planta Agrobacterium- mediated transformation of L-Asparaginase gene into potato ( Solanum tuberosum L.) cv. Agria: an efficient and novel method

Since potato is the fourth most important crop, the development of a practical and simple transformation method for cultivars recalcitrant in tissue culture is absolutely vital. When the potato is fried at temperatures higher than 120 °C, acrylamide that is probably carcinogenic in humans is formed in French fries and potato chips during the Maillard reaction. An effective solution to diminish the level of acrylamide is to transform potato cultivars by overexpressing the L-Asparaginase gene in potato tubers. In this study, a vector was constructed with the L- Asparaginase gene downstream of the laccase tuber-specific promoter using the expression vector pBI121. Agrobacterium tumefaciens suspension was injected into the eyes of potato tubers using insulin syringes. Out of all selected plants, 127 putative transgenic lines were resistant to 0.5% glyphosate, and the mean transformation efficiency turned out to be 31.75%. This is the first report of in planta transformation utilized for potato. The expression of the transgene in the tubers of transgenic lines at RNA and protein level was validated by RT-PCR and ELISA respectively. The recombinant enzyme containing a 6xHistidine-tag was easily purified by Ni–NTA chromatography and its molecular weight was determined to be 40.18 kDa by SDS-PAGE.