石斛属植物茎部总RNA提取方法的研究  

Studies on Total RNA Extraction Methods from the Stems of Dendrobium Species

作　　者：单婷婷 陈晓梅[1] 郭顺星[1] 王倩清 王爱荣[1] SHAN Ting-ting;CHEN Xiao-mei;GUO Shun-xing;WANG Qian-qing;WANG Ai-rong(Institute of Medicinal Plant Development, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100193;Agriculture College, Ludong Univercity, Yantai 264025)

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193 [2]鲁东大学农学院,烟台264025

出　　处：《生物技术通报》2018年第6期54-58,共5页Biotechnology Bulletin

基　　金：国家自然科学基金项目(81573594);中国医学科学院医学与健康科技创新工程(2017-I2M-3-013)

摘　　要：获得快速提取高质量石斛属植物茎总RNA的方法,为深入开展分子生物学研究奠定基础。采用7种方法提取铁皮石斛茎总RNA,凝胶电泳和紫外分光光度法检测质量,筛选最佳方法,并对方法进行验证。结果显示,经过比较,用改良华越洋多糖多酚试剂盒法(M7)提取的铁皮石斛茎总RNA的完整性、浓度和纯度均最好。用M7提取铁皮石斛、金钗石斛、鼓槌石斛、球花石斛和重唇石斛的茎总RNA,以及提取不同生长条件(温室和大棚种植)和不同生长时间(大棚生长3个月、1年和2年)铁皮石斛的茎总RNA,所获样品的完整性、浓度和纯度均符合质量要求。M7操作简便,结果重复性好,适于石斛属植物茎总RNA的提取。This work is aimed to establish a fast and efficient method of extracting total RNA from the stem of Dendrobium species for further molecular biology investigation. Seven methods were used to extract the total RNA from the stem of Dendrobium officinale,then gel electrophoresis and UV spectrometer were applied to determine the quality of the extracted total RNA,and the screened method was verified by samples of D. officinale of different growth conditions and times,and of 5 species of Dendrobium. Results showed that the integrity,purity and extraction capacity of the stem total RNA of D. officinale by Improved HUAYUEYANG Quick RNA Isolation Kit（（M7）was the best. The stem total RNA,extracted using M7,of D. officinale,of D. nobile,of D. chrysotoxum,of D. thyrsiflorum and of D. hercoglossum,and of D. officinale planting in plastic house and greenhouse for 1 year respectively,as well as in plastic house for 3,12 and 24 months respectively,were verified to meet the quality requirements on the integrity,purity and extraction capacity. In conclusion,M7 is suitable for stem total RNA extraction of Dendrobium species for easy handle and fine repeatability.

关 键 词：石斛属植物 总RNA提取 改良华越洋多糖多酚试剂盒法 

分 类 号：S567.239[农业科学—中草药栽培]