Optimized Regeneration of Petunia Protoplast and Its Association with Tissue Identity Regulators

The popular ornamental plant Petunia is also a valuable model in tissue culture. Cellular conversions during differentiation and regeneration have been investigated using various combinations of phytohormones; however, studies on genes for reprogramming toward desired identities limited. In this study, we isolated protoplasts cultured them the callus, rooting, or shooting stages, which were used to establish optimal protoplast culture conditions identify that epigenetically function as identifiers. plasmolysis enzyme digestion obtain healthy compared, Viscozyme, Celluclast, Pectinex (VCP) enzymes more efficient isolating when followed by 21 25% sucrose purification washing processes. filtered washed started divide at 1 day developed into colonies after 3 weeks culture, showed higher efficiency Murashige Skoog (MS) salt media compared Kao Michayluk (KM) media. pluripotent formed calli solid medium supplemented with 3% 4 weeks, destined same cell mass, medium. Three epigenetic controllers, ATXR2, ATX4A, ATX4B, highly expressed calli, shoots, organs shoots roots, respectively, confirming dedifferentiation identity plastic.