Estandarización del protocolo de transformación genética de células embríogénicas de banano de la variedad ‘williams’ (aaa) mediada por agrobacterium tumefaciens

Abstract

A conventional breeding strategy in banana is difficult due to the high level of sterility, need of interploidy crosses, poor seed germination rate and long life cycle. Genetic engineering is an alternative to conventional breeding and offers plant breeders an opportunity to overcome the constraints imposed by sterility of most banana cultivars. The principal objective of this study is the development and standardization of methodologies for the integration of genes into the banana genome in Ecuador. The transfer of genes was performed through the co-cultivation of embryogenic cell suspensions of the banana cultivar ‘Williams’ with Agrobacterium tumefaciens strain EHA105. The plasmids used for transformation were the pCAMBIA1301 (35S promoter driving the uidAINT reporter gene) and the pESKUL1 and pESKUL7 which both contain different banana promoters driving the uidAINT reporter gene. Transient gene expression assays showed up to 12, 26 and 244 blue foci in banana cells transformed with pESKUL1, pESKUL7 and pCAMBIA1301, respectively. The optimal concentration of the antibiotic hygromycin was tested for the selection of transgenic cell colonies. A standardized protocol for genetic transformation in banana is been developed, setting the basis for the generation banana plants resistant to diseases for the first time in Ecuador.