Efficient Transformation of Solanum tuberosum cv. Spunta Mediated by Agrobacterium tumefaciens Using Hygromycin as a Selective Agent

Abstract

Plant genetic transformation is a biotechnological tool with a high impact on plant breeding, since it offers the possibility of introducing new traits in crops to fulfill the demands of production and consumption. The most widely used transformation strategy in potatoes is based on the integration of genes via A. tumefaciens. The objective of the present work was to develop a transformation protocol for Solanum tuberosum cv. Spunta mediated by A. tumefaciens using hygromycin as a selective agent. For this, explants of potato cv. Spunta were transformed using A. tumefaciens with a binary vector containing the hygromycin resistance transgene (hpt) under the control of the NOS promoter. Molecular and phenotypic determinations were performed to validate both the efficiency of the transformation protocol and the performance of hygromycin as a selection agent. Sixty-two regenerated shoots were obtained from explants inoculated with A. tumefaciens (62% regeneration), of which 30 plants were positive for the hpt gene. A thirty percent transformation efficiency, calculated as the number of transgenic plants / the number of explants inoculated, was obtained using this protocol. The present work presents a simple and efficient protocol for the genetic transformation of potato cv. Spunta by A. tumefaciens using hygromycin as a selective agent.