Efficient protocol improved the yield and viability of oil palm protoplasts isolated from in vitro leaf and mesocarp

• An efficient protocol for protoplasts isolation from in vitro leaf and mesocarp of oil palm has been established. Enzymes composition effect on the yield viability protoplast are significant. Scraping waxy layer increased efficiency. Vacuum treatment enhanced penetration enzymes into tissues. 10% ficoll 0.6 M mannitol improved separation impurities. The absence a cell wall contributes to its versatility. Its flexibility DNA manipulation possibility rapid cell-based assay is desirable plant biotechnology field. This study was carried out improve factors affecting efficiency were optimized, including protocols enzyme involved, focusing first. Incubation sample with an mixture cellulase R-10, macerozyme driselase, pectolyase Y-23, 14 h successfully produced up 2.5 × 10 6 g -1 fresh weight (FW) 95% viability. tissue optimized 2 at static condition also 3.98 FW 85% Besides, it found that increasing sample's surface area contact solution by slicing samples narrow strips thin layers tissues In addition, plasmolysis step before enzymatic minimizing damage incurred during isolation. successful enabled gene function characterization endogenous tissue-specific promoter being vivo .