安徽农业科学2011，Vol.39Issue(35)：21644-21646,3.
药用植物青天葵叶片原生质体的制备与纯化
Preparation and Purification of Protoplast from Nervilia fordii (Hance) Schltr.Leaves
摘要
Abstract
[Objective] To study the preparation and purification method of protoplast from Nervilia fordii (Hance) Schltr. Leaves. [Method] Taking the yield and survival rate of protoplast as index, the effects of different factors such as pretreatment, enzymolysis method, enzymolysis temperature, rotate speed of centrifugation, illumination, and sieve mesh number on the separation and purification of protoplast from N. Fordii leaves were studied. [Result] The optimum process for protoplast isolation was obtained as follows; N.fordii leaves were first pretreated for 1 h in CPW solution containing 13% mannitol,and then incubated at (25 ± 1)℃ in enzyme solution under dark conditions; after the enzymolysis,the enzyme liquid was filtrated through 200 mesh screen and centrifuged for 8 min at the speed of 800 r/min. Repeat the filtration and centrifugation once before microscopic examination. Under the above conditions, a large amount of complete and clear protoplast with few cell debris was obtained. [ Conclusion ] The preparation system has been successfully established and laid a good foundation for establishing protoplast regeneration system from N. Fordii leaves.关键词
青天葵(LEAF OF FORD NERVILIA)/叶片/原生质体(protoplast)/分离Key words
LEAF OF FORD NERVILIA/ Leaf/ Protoplast/ Isolation分类
医药卫生引用本文复制引用
张晓丽,何瑞,童家赟,詹若挺,林超..药用植物青天葵叶片原生质体的制备与纯化[J].安徽农业科学,2011,39(35):21644-21646,3.基金项目
国家自然科学基金(30701090) （30701090）
教育部留学回国人员科研项目(教外司留[ 2009] 1001号). （教外司留[ 2009] 1001号）
