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Position: Home > Articles > Isolation conditions of protoplast from Nervilia fordii(Hance) Schltr. leaf Guangdong Agricultural Sciences 2011,38 (19) 3+107-108+131

青天葵叶片原生质体分离条件的优化

作  者:
张晓丽;何瑞;童家赟;詹若挺;林超
单  位:
广州中医药大学中药资源科学与工程研究中心/省部共建中药资源科学教育部重点试验室
关键词:
青天葵;叶片;原生质体;分离
摘  要:
以青天葵Nervilia fordii(Hance)Schltr.的新鲜叶片为试材,对青天葵原生质体的制备分离技术进行研究。用不同浓度的甘露醇溶液处理青天葵叶片下表皮细胞,进行细胞质壁分离试验,确定青天葵叶片细胞的等渗浓度;采用正交设计研究酶解法制备青天葵叶片原生质体的最适合纤维素酶浓度、离析酶浓度及酶解时间。结果表明,青天葵叶片细胞的等渗浓度为11%甘露醇;采用1.0%纤维素酶R-10+0.6%离析酶R-10的混合酶液酶解青天葵叶片12 h能达到很好的分离效果,获得高质量的原生质体。
译  名:
Isolation conditions of protoplast from Nervilia fordii(Hance) Schltr. leaf
作  者:
ZHANG Xiao-li1,HE Rui1,TONG Jia-yun1,2,ZHAN Ruo-ting1,LIN Chao1(1.Research Centre of Chinese Herbal Resource Science and Engineering/Guangzhou University of Chinese Medicine, Guangzhou 510006,China;2.College of Chinese Traditional Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510006,China)
关键词:
Nervilia fordii;leaf;protoplast;isolation
摘  要:
Leaf were used as starting material to optimize the isolation conditions of Nervilia fordii(Hance) Schltr.protoplast.The lower epidermis of Nervilia fordii leaf was treated with different concentration of mannitol solution to determine the optimal osmotic pressure.The cellulase concentration,macerozyme concentration,and hydrolysis time for the separation of protoplasts were studied by orthogonal design.The results showed that the isotonic concentration of mannitol for Nervilia fordii leaf cells was shown to be 11%.The highest yield of protoplast was obtained using the mixed enzyme solution containing 1.0% cellulase R-10+0.6% macerozyme R-10 and incubated for 12 hours.

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