In vitro enzymatic isolation of protoplasts from tissues of the medicinal plant Physalis angulata L.

Physalis angulata is a medicinal plant from the Solanaceae family. Utilization of tissue culture, especially protoplast techniques, has potential for supplying medicinal plants in the framework of their development as standardized herbal medicine. Plant protoplasts can be enzymatically isolated from tissues culture. The composition of enzyme solutions for isolating protoplasts is critical to obtain high numbers of viable protoplasts. The length of incubation during the isolation process also determines the quantity and quality of the released protoplast. Each tissue source may require special conditions for successful isolation; therefore, the objective of this study was to observe the effect of enzyme solution and duration of incubation on the number of viable protoplasts released from P. angulata tissues in vitro. Five treatments for protoplast isolation combining cellulose and macerozyme were tested in vitro on leaf, stem and callus tissues as protoplast donors. Supplementing with an osmoticum (0.6 M mannitol) and a membrane stabilizer (10 mM CaCl2) provided significantly different numbers of viable protoplasts. In this study, all isolation conditions could not liberate protoplasts from callus tissues. The tested concentration of osmoticum only facilitated callus cells to plasmolyze. Spherical protoplasts containing distributed plastids were still inside the individual cells after 5 h incubation. Leaf protoplasts showed the most exciting results. The highest number of viable protoplasts (1.54 × 105 protoplasts/mL) was obtained from mesophyll tissues after 2 h incubation. The content of high-density green chloroplasts was a striking feature of leaf protoplasts. This feasible method to obtain leaf protoplasts from P. angulata is an excellent prospect for fusion experiments.Physalis angulata is a medicinal plant from the Solanaceae family. Utilization of tissue culture, especially protoplast techniques, has potential for supplying medicinal plants in the framework of their development as standardized herbal medicine. Plant protoplasts can be enzymatically isolated from tissues culture. The composition of enzyme solutions for isolating protoplasts is critical to obtain high numbers of viable protoplasts. The length of incubation during the isolation process also determines the quantity and quality of the released protoplast. Each tissue source may require special conditions for successful isolation; therefore, the objective of this study was to observe the effect of enzyme solution and duration of incubation on the number of viable protoplasts released from P. angulata tissues in vitro. Five treatments for protoplast isolation combining cellulose and macerozyme were tested in vitro on leaf, stem and callus tissues as protoplast donors. Supplementing with an osmoticum (0.6 M m...