Agrobacterium - Mediated Transformation of Secondary Somatic Embryos from Rosa Hybrida L. and Recovery of Transgenic Plants (original) (raw)

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 PCRand 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.

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