一种改良的杧果叶片总RNA提取方法  

A Modified Method of Total RNA Isolation for Mango Leaf Tissues

作　　者：黄章保 任惠[1] 董龙[1] 蔡昭艳[1] 方位宽[1] 王小媚[1] 苏伟强[1] 刘业强[1] 邱文武[1] Huang Zhangbao;Ren Hui;Dong Long;Cai Zhaoyan;Fang Weikuan;Wang Xiaomei;Su Weiqiang;Liu Yeqiang;Qiu Wenwu(South Asian Tropical Fruit Tree Scientific Observation Station(Nanning),Ministry of Agriculture and Rural Affairs,Horticultural Research Institute,Guangxi Academy of Agricultural Sciences,Nanning,530007)

机构地区：[1]广西壮族自治区农业科学院园艺研究所农业部南宁南亚热带果树科学观测实验站,南宁530007

出　　处：《分子植物育种》2021年第12期3983-3989,共7页Molecular Plant Breeding

基　　金：广西重点研发计划项目(桂科AB18294031);广西重点研发计划项目(桂科AB16380068);广西农业科学院基本科研业务专项项目(桂农科2020YM28;桂农科2015JZ82);广西农业科学院基本科研业务专项优势学科团队项目(桂农科2015-YT47)共同资助。

摘　　要：杧果叶片组织复杂的代谢成分导致RNA提取难度较高。为了提高总RNA提取质量,本研究在已有文献报道方法的基础上,改进并获得了一种利用丙酮洗涤液氮研磨材料和使用0.3 mol/L氯化锂(LiCl)溶液、2倍体积无水乙醇作为第二次沉淀剂的RNA提取方法。实验结果表明,改进方法显著减少了RNA提取过程中的杂质沉淀量,提取的总RNA电泳条带完整,OD260/280为1.9左右,平均得率达到329.8 g/g FW±11.2μg/g FW。RNA反转录和双链c DNA相关序列多态性扩增(SRAP-PCR)的进一步验证实验获得了清晰的多态性条带,表明提取的杧果叶片总RNA具备较高质量,适用于开展分子生物学实验。Complex metabolic components in mango leaves lead to high difficulty in RNA extraction.In order to improve the quality of total RNA extraction,on the basis of the existing methods reported in the literature,an RNA extraction method combining acetone washing liquid nitrogen abrasive material with 0.3 mol/L lithium chloride(LiCl)solution and 2 times the volume of anhydrous ethanol as the second precipitator was developed.The experimental results showed that the improved method significantly reduced the amount of impurity precipitation in the RNA extraction process.The electrophoretic bands of total RNA extracted were complete.The absorbance ratio of OD260/280 was about 1.9,and the average was 329.8 g/g FW±11.2 g/g FW.Further validation experiments of RNA reverse transcription and the polymorphic amplification of double-stranded cDNA related sequences(RAP-PCR)obtained clear polymorphic bands,showing that the total RNA from mango leaves had high quality and was suitable for molecular biology experiments.

关 键 词：杧果 RNA提取方法 多糖 多酚 

分 类 号：S667.7[农业科学—果树学]