Plasma membrane of plant cells is surrounded by cellulose wall and adjacent cells are joined together by a thick pectin rich matrix. Separation of plant cells and removal of the cell wall experimentally, by either a m...Plasma membrane of plant cells is surrounded by cellulose wall and adjacent cells are joined together by a thick pectin rich matrix. Separation of plant cells and removal of the cell wall experimentally, by either a mechanical or an enzymatic process, results in the production ofprotoplast. Protoplasts are useful tools to study the uptake and transport ofmacromolecules and production of somatic hybrids. Protoplasts can be obtained from all types of actively growing young and healthy tissues. The most convenient and widely used source of plant protoplasts is leaf. Juvenile seedling tissues, cotyledons are other alternative tissues most frequently used for protoplasts isolation. All the environmental and genotypic factors, which affect the cell wall thickenings and compactness indirectly, influence the number of protoplasts recovered. Protoplasts are isolated by two methods, mechanical and enzymatic. The enzyme mixture solution of celluiose/macerozyme is used to digest the cell wall. The critical factors affecting the obtaning ofprotoplasts are the kinds of cell wall degrading enzymes, the physiological state of plant leaves, the type of osmotic stabilizers and the composition of reaction solution. With the improvement of technique and enzyme combination rate, the yield of collected protoplasts will be increased higher.展开更多
文摘Plasma membrane of plant cells is surrounded by cellulose wall and adjacent cells are joined together by a thick pectin rich matrix. Separation of plant cells and removal of the cell wall experimentally, by either a mechanical or an enzymatic process, results in the production ofprotoplast. Protoplasts are useful tools to study the uptake and transport ofmacromolecules and production of somatic hybrids. Protoplasts can be obtained from all types of actively growing young and healthy tissues. The most convenient and widely used source of plant protoplasts is leaf. Juvenile seedling tissues, cotyledons are other alternative tissues most frequently used for protoplasts isolation. All the environmental and genotypic factors, which affect the cell wall thickenings and compactness indirectly, influence the number of protoplasts recovered. Protoplasts are isolated by two methods, mechanical and enzymatic. The enzyme mixture solution of celluiose/macerozyme is used to digest the cell wall. The critical factors affecting the obtaning ofprotoplasts are the kinds of cell wall degrading enzymes, the physiological state of plant leaves, the type of osmotic stabilizers and the composition of reaction solution. With the improvement of technique and enzyme combination rate, the yield of collected protoplasts will be increased higher.
