Production of transgenic creeping bentgrass Agrostis stolonifera var. palustris plants by Agrobacterium tumefaciens-mediated transformation using hygromycin selection

Abstract

A protocol was developed for Agrobacterium tumefaciens-mediated transformation of creeping bentgrass [Agrostis stolonifera L. var. palustris (Huds) Farw]. The transformation was performed using the vector pCAMBIA 1301 which contains the reporter (uidA) gene and the selectable marker hygromycin phosphotransferase (hph) gene. Embryogenic calli initiated from mature seeds were infected with A. tumefaciens strain EHA105 followed by hygromycin selection. Effects of some variables influencing efficiency of T-DNA delivery and selection were examined, and the protocol was successfully employed in the production of transgenic creeping bentgrass. Southern blot analysis demonstrated the integration of a single or two copies of foreign gene, and RT-PCR confirmed the transcription of the genes.