葱属主要蔬菜作物高质量RNA提取方法研究  

High Quality RNA Extraction Method from Main Allium Vegetables

作　　者：孙亚玲[1,2] 陈立[1,2] 刘冰江[1] 缪军[1] 孔素萍[1] 高莉敏[1] 曹辰兴[2] 吴雄[1] 杨妍妍[1] 霍雨猛[1] 

机构地区：[1]山东省农业科学院蔬菜花卉研究所/山东省设施蔬菜生物学重点实验室/国家蔬菜改良中心山东分中心,山东济南250100 [2]山东农业大学园艺科学与工程技术学院,山东泰安271018

出　　处：《山东农业科学》2016年第9期6-9,16,共5页Shandong Agricultural Sciences

基　　金：国家自然科学基金项目(31201635;31301786);山东省农业科学院青年英才重点扶持项目

摘　　要：葱属蔬菜作物(洋葱、大葱、大蒜和韭菜等)各组织部位富含多糖等次生代谢物质,严重影响了RNA的提取质量。本研究以洋葱叶片为样本,对多种RNA提取方法进行了比较分析,获得了一套适合葱属蔬菜作物RNA提取的系统方法。该方法将CTAB-GT连用、蛋白酶K替代DEPC和冰浴电泳检测相结合,其所提取的RNA样本产量约为152.35μg/g FW,RNA完整性RIN值约为7.5,条带清晰,无多糖、蛋白质和DNA残留,28S约为18S两倍,提取过程安全无强致癌物质DEPC,可满足下游RNA分子实验的要求。且该方法适合于大葱、洋葱、大蒜和韭菜不同组织部位样本RNA的提取。Allium vegetables（ including Allium cepa,Allium fistulosum L. var. giganteum Makion,Allium sativum and Allium tuberosum Rottl. ex Spreng） are rich in polysaccharides and other secondary metabolites,which seriously affect the quality of extracted RNA. In this study,we compared and analyzed a variety of RNA extraction methods using onion leaves as samples,and obtained a set of method for RNA extraction from Allium vegetables. This method was comprised of CTAB-GT,Proteinase K instead of DEPC and ice bath gel electrophoresis. The yield of RNA samples was about 152. 35 μg / g FW,the RNA integrity（ RIN value） was about 7. 5,the bands of gel electrophoresis were clear without contaminations of polysaccharides,protein and DNA residues,the ratio of 28 S /18 S was about 2 and the extraction process was safe without strong carcinogen of DEPC. The RNA quality was suitable for downstream molecular experiments. Besides,this method could be applied to the extractions of different tissues of welsh onion,onion,garlic and Chinese chives.

关 键 词：葱属蔬菜 RNA提取 多糖 CTAB-GT 

分 类 号：S633[农业科学—蔬菜学]