新疆雪莲胚性愈伤组织的诱导及原生质体分离  

Studies on Embryogenic Callus Induction and Protoplast Isolation of Saussurea involucrate Kar et Kir.

作　　者：袁永娴[1,2] 王晓军[1] 郝秀英[3] 赵民安[1] 朱金刚[1,2] 祁鑫星[1,2] 牛力涛[1,2] 

机构地区：[1]中国科学院新疆理化技术研究所,乌鲁木齐新疆830011 [2]中国科学院研究生院,北京100039 [3]新疆农业科学院微生物应用研究所,乌鲁木齐新疆830091

出　　处：《北方园艺》2010年第8期125-128,共4页Northern Horticulture

基　　金：中国科学院西部行动高新技术资助项目(KGCX2=YW-509)

摘　　要：以新疆雪莲无菌叶片为材料，诱导其产生胚性愈伤组织，再以胚性愈伤组织为分离原生质体的起始材料，研究质壁分离时间、渗透压浓度、酶液组合、酶解时间等因素对其原生质体分离效果的影响。结果表明：对诱导胚性愈伤组织有较好效果的培养基组合为：MS＋2，4-D0．1mg／L＋BA0．5mg／L＋NAA1．5mg／L＋葡萄糖30g／L；原生质体分离最佳体系为：含纤维素酶2．5％、离析酶0．2％、果胶酶1％、IVIES0．1％和甘露醇10％的酶解液，在温度（26±1）℃，pH5．8条件下酶解14～16h，游离原生质体的产量和活性均较高。The leaves of Saussurea involucrate were used as experiment materials and were induced embryogenie callus, then using them to isolate protoplast. The influential factors of plasmolysis time, concentration of osmotic solution, enzyme combinations and digestion time on protoplast isolation from embryogenic callus were studied. It is suitable to indicate embryogenic calluses when the mttrashige and skoog medium which was MS＋2,4-D 0. 1 mg/L＋BA 0. 5 mg/L＋ NAA 1.5 mg/L＋glucose 30 g/L. When the embryogenic calluses were used to isolation, the optimal condition was enzyme solution of 2. 5% Cellulase Onzuka R-10,0. 2% Mecerozyme R-10,1.0% Pectinase,0. 1% MES and 10%Mannitol for digesting 14-16 h at pH 5. 8 and the temperature of（26±1）℃.

关 键 词：新疆雪莲 胚性愈伤组织 原生质体 分离 

分 类 号：S567.23[农业科学—中草药栽培]