园艺植物生物技术第四章题库.ppt

Basic procedures of enzymolysis method Culture methods Plate culture 平板培养 Nurse culture看护培养 Microculture微室培养 Conditioned medium culture条件培养基培养 The cells in culture exhibit the following five phases of a growth cycle i. Lag phase, where cells prepare to divide. ii. Exponential phase, where the rate of cell division is highest. iii. Linear phase, where cell division slows but the rate of cells expansion increases. iv. Deceleration phase, where the rates of cell division and elongation decreases. v. Stationary phase, where the number and size of cells remain constant. 2. Protoplast Isolation and Fusion原生质体的游离与融合 2.1. Protoplast Isolation Methods mechanical methods enzymatic methods 2.1.1 Mechanical method Application range large and highly vacuolated cells of storage tissues onion bulb scales, radish root and beet root tissue Disadvantages (i) It is restricted to certain tissues which have large vacuolated cells. (ii) Yield of protoplasts is generally very low. Protoplasts from less vacuolated and highly meristematic cells do not show good yield. (iii) The method is tedious and laborious. (iv) Viability of protoplasts is low because of the presence of substances released by damaged cells. 2.1.2 Enzymatic method Cocking在1960年证实了用酶从高等植物中分离出原生质体的可能性。 Cocking in 1960 demonstrated the possibility of enzymatic isolation of protoplasts from higher plants. Influencing factors (1)Physiological state of tissue and cell material Protoplasts have been isolated from a variety of tissues and organs including leaves, petioles, shoot apices, roots, fruits, coleoptiles, hypocotyls, stem, embryos, microspores, callus and cell suspension cultures of a large number of plantspecies. 分离材料包括叶片、叶柄、芽尖、根、果实、胚芽鞘、胚轴、茎、胚芽、花粉粒、愈伤组织和细胞悬浮培养物等。植物幼苗或新生枝的完全伸展叶片的叶肉组织是分离原生质体的最方便、最合适的植物材料。 (2) Enzymes The release of protoplasts is very much dependent on the nature and composition of enzymes used to digest the cell wall. 原生质体的释放在很大程度上取决于用于消化细胞壁的酶的性质和组成。 Three primary components