何首乌不同部位二苯乙烯苷含量以及芪合酶FM-STS时空表达差异  

The Content of Stilbene Glycosid in Different Parts of Polygonum Multiflorum Thunb and the Stilbene Synthase FM-STS Spatial and Temporal Expression

作　　者：朱宽鹏[1,2] 生书晶[1,2] 赵炜[1,2] 陆娣[1,3] 夏晚霞[1,2] 赵树进[1] 

机构地区：[1]广州军区广州总医院,广东广州510010 [2]华南理工大学生物科学与工程学院,广东广州510640 [3]南方医科大学,广东广州510515

出　　处：《现代生物医学进展》2012年第30期5819-5822,5804,共5页Progress in Modern Biomedicine

基　　金：广东省自然科学基金(10151001002000012);广东省科技计划(00697862100303016)

摘　　要：目的:探究何首乌不同部位主要有效成分二苯乙烯苷含量差异以及相对应部位何首乌芪合酶基因FM-STS表达差异。方法:通过液氮碾磨提取广东德庆同一株何首乌根茎叶中的RNA以及用50%稀乙醇过夜浸提芪合物二苯乙烯苷;使用乙腈和水为流动相(体积比为20∶80),利用高效液相色谱分析其二苯乙烯苷含量差异;采用实时荧光定量PCR分析芪合酶基因Fm-STS表达差异,以β-actin作为内参对照,2-△△CT公式计算各组别中FM-STS的含量。结果:根茎叶中芪合物二苯乙烯苷含量依次为14.62mg/g、1.78mg/g和0.47mg/g(干重),在mRNA水平上检测基因Fm-STS表达量为叶片中最高,根是叶的1/10倍,茎是叶的1/64倍。结论:芪合酶基因FM-STS主要在何首乌叶片中表达,二苯乙烯苷主要在叶片中合成,进而转移到根块中富集。Objective： To investigate the content of main active ingredient- Stilbene glycosid in differet parts of Polygonum Multiflornm Thunb and the stilbene synthase gene FM-STS expression differences in its corresponding parts. Methods： Using liquid nitrogen to mill root, stem and leaves at one Polygonum Multiflorurn Thunb obtained from Guangdong Deqing, extracted the total RNA and using 50 % dilute ethanol to extract stilbene glycosides, using acetonitrile and water as the mobile phase （volume ratio 20：80） in High Performance Liquid Chromatography to analyz the contents difference; Real-time PCR was used to analyze the stilbene synthase gene Fm-STS expression differences and 13-actin was used as internal controls, 2 .~cr formula was used to calculate each category FM -STS content. Result： The content of Stilbene glycosid in root/stem/leaf were 14.62mg/g, 1.78mg/g and 0.47mg/g （dry weight）, respectively. The gene Fm-STS expression at the mRNA level is the highest, and it was 10 times as that in root＇s and 64 times as that in stem＇s. Conclusion： Stilbene synthase gene FM-STS expressed mainly in the Polygonum Multiflorum Thunb leaves, and stilbene glueoside synthesized in leaves, and then transferred to the root block for enrichment.

关 键 词：何首乌 二苯乙烯苷 芪合酶基因 荧光定量PCR 

分 类 号：R284.2[医药卫生—中药学] Q943[医药卫生—中医学]