Ribosome Inactivating Protein of barley enhanced resistance to Rhizoctonia solani in transgenic potato cultivar 'Desirée' in greenhouse conditions

In the present study, the potato cultivar 'Desirée' was transformed via Agrobacterium tumefaciens strain LBA4404 containing the plasmid pBIN19 which harbors the Ribosome Inactivating Protein (rip30). The potato leaf discs were used as an explant for transformation. The in vitro regeneration parameters (percentage of callus regenerated, number of shoots per callus, percentage of regenerated roots and percentage of the transgenic plants) were evaluated. The PCR technique was used for identification of transformed plants. Southern and Western blot analyses were applied for molecular characterization of the transgenic clones. A greenhouse assay was carried out to evaluate the resistance to Rhizoctonia solani pathogen of transgenic clones expressing the rip30 gene. The results revealed that not all the plants developed in selective medium were positive for the corresponding gene using the PCR technique. Southern blot analysis demonstrated that the tested transgenic plants integrated three copies of rip30 gene into their genome. The expression of the RIP30 protein was confirmed in the leaf extracts of the transgenic clones by Western blot analysis. Resistance evaluation of the transgenic plants in greenhouse conditions showed that disease incidence and severity were reduced for R. solani.