Agrobacterium - mediated transformation of secondary somatic embryos from rosa hybrida l. And recovery of transgenic plants

Abstract

A genetic transformation protocol, based on co-cultivation of Rosa hybrida L. secondary embryos with Agrobacterium was established. Transgenic phosphinothricin (ppt) - resistant plants were obtained through application of interrupted selection procedure. Following the Agrobacterium inoculation step, the somatic embryos were initially cultivated on ppt—free media containing cefatoxim for elimination of the bacteria and later transferred on media containing the selective agent. The ppt—resistant secondary somatic embryos were matured and regenerated on ppt—free media and the putative transgenic plants were subsequently selected on media contained ppt and chlorphenol red. The performed PCR—and PAT protein—assays demonstrated presence and expression of bar gene encoding the phosphinothricin acetyltransferase (PAT) in part of the selected plants. The possibilities for application of the established transformation protocol are discussed. © 2005 Taylor and Francis Group, LLC.