枣树原生质体分离条件的研究

A Study of the Separation Conditions for Protoplast Separating of Zizyphus jujuba

枣树原生质体分离研究是其原生质体培养与融合、外源遗传物质转化等研究的基础工作．用胚性悬浮培养细胞、细粒状胚性愈伤组织、未细切愈伤组织和已细切愈伤组织等４种材料进行原生质体分离．结果表明，枣树胚性悬浮培养细胞是最佳起始材料，用浓度为１０ｇ／Ｌ纤维素酶＋１ｇ／Ｌ离析酶＋ＣＰＷ盐（１３２０ｍｇ／ＬＣａＣｌ２·２Ｈ２Ｏ和１００ｍｇ／ＬＫＨ２ＰＯ４·Ｈ２Ｏ组成的混合液）＋０．７ｍｏｌ／Ｌ甘露醇组成的混合酶液对起始材料进行１６ｈ的酶解，可获得较高的原生质体产量，且原生质体活力较高．

With such four materials as embryonic suspension cultivated cells,fine granular embryonic callus,un cutting callus and sliced callus,the protoplast of Zizyphus jujuba was separated.The separation results show that:Embryonic suspension cultivated cells are the best initiation materials for protoplast isolation.When they were decomposed for 16 hours in an enzyme mixture solution containing 1% of onozuka,0.1% of pecitinase,and 0.7 mol/L of mannital CPW(1 320 mg/L CaCl 2·2H 2O and 100 mg/L KH 2PO 4·H 2O),the optimum yield and viability of protoplast were abtained.