流式细胞法测定茅苍术基因组大小  

Flow Cytometric Analysis of Genome Size in Atractylodes lancea

作　　者：倪金菁 贺彬[2] 汪文杰[1] 余坤[1] 

机构地区：[1]湖北中医药大学药学院,湖北武汉430065 [2]湖北中医药大学基础医学院,湖北武汉430065

出　　处：《中药材》2015年第6期1153-1156,共4页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金(31300277);高等学校博士学科点专项科研基金(20124230120002);湖北省教育厅优秀中青年人才项目(Q20121611);湖北中医药大学校级课题

摘　　要：目的:建立流式细胞术测定茅苍术基因大小的方法,并进一步比较5种类型茅苍术的基因组大小变异情况。方法:以茅苍术幼叶为材料,采用改良Otto两步法提取细胞核,用已知基因组大小的玉米和蚕豆为内标,将处理好的样品上流式细胞仪测定。结果:通过改进实验方法,以蚕豆为内标,建立了流式细胞术测定茅苍术基因组大小的方法。5种类型茅苍术基因组大小的测定结果表明,尖叶类型基因组最大,扁茎类型最小,裂叶类型的基因组大小稍有下降。结论:本研究为茅苍术基因组大小研究提供了方法和数据参考,也将为茅苍术群体变异、种质资源评价和新品种选育等研究提供基础。Objective ： To develop a rapid analytical method for the determination of genome size of Atractylodes lancea by flow cy- tometry（ FCM）, and to estimate genome size of five typical plant types of cultivated Atractylodes lartcea. Methods ： The fresh young leaves of Atractflodes lancea were used for the preparation of nuclear suspension with two-step protocol. After staining with propidium iodide, the mixture was analyzed by flow cyt0metry. Zea mays ＇ CE-777＇ or Vicafaba ＇ Inorce＇ was used for DNA reference standard. Results： The flow cytometric method was developed after screening internal standard, optimizing sample preparation and FCM setups. The narrow leaf type of Atractylodes lancea got the biggest genome size, whereas the smallest one was fasciated stem type. It was suggested that parted leaf might be caused by genome size decrease. Conclusion：The methodology presented in this study is suitable for measuring the genome size of Atractylodes lancea. This study also provides useful information on population variation, evaluation of germplasm re- sources, and breeding of Atractylodes lancea.

关 键 词：茅苍术 基因组大小 流式细胞术 C值 

分 类 号：R282.71[医药卫生—中药学]