Fig. 3

Schematic illustration of areca palm protoplast isolation. a A healthy 18-month-old areca palm plant suitable for protoplast isolation. Dashed-line box indicates the optimal leaflets (light green leaflet of the newly emerged leaf) to be used as the mesophyll protoplast source; b Selected leaflet was cut into 2–4 cm long for peeling; c Areca palm leaflet after peeling the waxy surface using a sharp pointed tweezer; d Peeled leaflet sections incubated in the enzyme solution for releasing protoplasts; e Protoplasts released in enzyme solution after overnight incubation; f Green solution containing released protoplasts in a petri dish; g Pelleting protoplast through optimized low-speed centrifugation; h Isolated areca palm protoplasts resting in MMg solution; i. Protoplast viability test using FDA staining