植物组织培养理论.ppt

Growth medium is optimised for regeneration of plantlets No sucrose (0%) Sucrose reduced to 1% Sucrose increased to 5% The explant is completely covered with green shoots, and roots are developing on the lower surface Very few shoots have developed on the explant. No roots and no callus. Shoots developed on edges of upper surface, and some root development has occured Shoots have developed over the surface of the explant, along with roots from the lower surface. The result is comparable with the control medium tissue culture in the African Violet No BAP? BAP reduced to 0.1 mg. l-1 No NAA Poor shoot development and no roots. Extensive callus generation Poor shoot development and no roots NAA reduced to 0.1 mg. l-1 More balanced development of roots and shoots. However, undifferentiated callus is developing at the edges of the explant Profuse development of roots and a reduced number of shoots. Undifferentiated callus is developing at the edges of the explant NAA increased to 5.0 mg. l-1 Profuse development of roots over the explant upper and lower surface, and few shoots. Some callus No MS salts No sign of regeneration from explant at all.? MS salts reduced to 0.47 g. l-1 ? Some root growth but reduced development of shoots MS salts increased to 9.52 g. l-1 ? Shoots developed on upper surface of explant. No roots or callus. 各种环境条件都会影响组织培养育苗的生长和发育。 温度 光照 湿度 培养基组成 pH值 渗透压 氧气 种类 最适pH值 种类 最适pH值 杜鹃 4.0 月季 5.8 越桔 4.5 胡萝卜、石刁柏 6.0 蚕豆 5.5 桃 7.0 番茄 5.7 ? ? 不同植物的最适pH值 【实验材料及仪器】 实验材料：胡萝卜 实验器材：蒸汽灭菌锅，超净工作台，酒精灯，三角瓶，镊子，剪刀，烧杯，培养皿 实验药品：MS培养基（见表一），蔗糖，琼脂，次氯酸钠，2.4D，NaOH 【实验步骤】 1.MS培养基母液的配置 按照表1配置MS培养基母液 培养基配制 培养基灭菌 材料消毒 接种 培养观察 愈伤组织分化 表一.MS培养基的配制 成分 母液 MS培养基 大量元素 NH4NO3 KNO3 CaCl2·H2O KH2PO4 MgSO4·7H2O mg/L(100×) 165000 190000 44000 17000 37000 mg/L 1650 1900 440 170 370 【实验步骤】 微量元素 mg/L(100×) mg/L MnSO4·4H2O ZnSO4·4H2O H3BO3 KI Na2MoO4·2H2O CuSO4·5H2O CoCl2·6H2O 2230 860 620 83 25 2.5 2.5 22.3 8.6 6.2 0.83 0.25 0.025 0.025 【实验步骤】 铁盐 Na2EDTA FeSO4·7H2O m