The effect of pre-cultivation of tobacco tissue culture on enzymatic separation of protoplasts from various cell types

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Abstract
A method of enzymatic separation of protoplasts from long-term tissue culture ofNicotiana tabacum L. is described. The efficiency of this method is dependent on conditions of separation and on the portion of meristematic cells in the tissue culture. This portion can be increased by pre-cultivations of the tissue on medium containing suitable concentration of hormones. The knowledge of the micromorphology of the filamentous culture enables us to investigate the course of release of protoplasts from various cell types. A preferential lysis of cell walls was observed between neighbouring cells in filaments and the fusion of their protoplasts was recorded. The preservation of cell walls which are not in a contact with other cells may be a result of the cell wall heterogeneity.

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