Protoplasts are plant cells lacking cell wall. Cell contents are surrounded by the cell membrane. They are a unique cell system to preform functional genetic analyses and studying wide specter of cell processes, for example cell structure, membrane function and hormonal signalization. Isolation of protoplast is a biotechnological technique for extracting protoplasts from plant tissue by enzymatic degradation of the cell wall. The success of isolation is influenced by various factors (physical, physiological ext.). The aim of the master's thesis was to check the success of protoplast isolation and viability of protoplasts of hemp (Cannabis sativa L.) by various procedures. We chose the most successful procedur and tried to optimize it. We determined which enzyme solution and from which plant parts the largest number of protoplasts were isolated. The results showed that the highest number of protoplasts can be isolated from mesophyll of leaf cells and etiolated hypocotyls. Few hours of incubation of leaves in enzyme solution composed of 1.5% Cellulase Onozuka R-10 and 0.4% of Macerozyme R–10, gave the best results. From etiolated hypocotyls the best results were obtained with enzyme solution composed of 1% Cellulase Onozuka R-10 and 0.1 % of Macerozyme R-10.
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