五种兜兰属植物基因组DNA提取方法比较  

Comparison of Genomic DNA Extraction Methods from Five Species of Paphiopedilum

作　　者：邵亚林 司俊波 常玮[2] 石家恋 丁勇[1] Shao Yalin;Si Junbo;ChangWei;Shi Jialian;Ding Yong(Key Laboratory for Forest Biotechnology in Yunnan,Department of Education,Southwest Forestry University,Kunming,650224;Key Laboratory of Economic Plants and Biotechnology,Kunming Institute of Botany,Chinese Academy of Sciences,Kunming,650204)

机构地区：[1]西南林业大学,云南省高校林木生物技术重点实验室,昆明650224 [2]中国科学院昆明植物研究所,资源植物与生物技术重点实验室,昆明650204

出　　处：《分子植物育种》2020年第15期4965-4974,共10页Molecular Plant Breeding

基　　金：国家自然科学基金(31400289);云南省大学生创新创业训练计划项目(201810677027);云南省高校林木生物技术重点实验室开放基金(51700201)共同资助。

摘　　要：为获得兜兰属植物基因组DNA的高质量提取方法,以麻栗坡兜兰(Paphiopedilum malipoense)、紫纹兜兰(Paphiopedilum purpuratum)、长瓣兜兰(Paphiopedilum dianthum)、硬叶兜兰(Paphiopedilum micranthum)和带叶兜兰(Paphiopedilum hirsutissimum)植物成熟叶片为试材,采用CTAB法、改良CTAB法、高盐低pH法和SDS法提取五种兜兰属植物基因组DNA,用琼脂糖凝胶电泳和分光光度计法分别检测所得总DNA的完整性和纯度。结果表明应用改良CTAB法从五种兜兰属植物中均可提取获得较高质量的基因组DNA,其它三种方法提取获得的兜兰属植物基因组DNA质量差异较大,CTAB法可从紫纹兜兰、长瓣兜兰和硬叶兜兰中提取获得较高质量的基因组DNA,SDS法可从麻栗坡兜兰和带叶兜兰中提取获得较高质量的基因组DNA。兜兰属植物基因组DNA的高质量提取方法获得为后续以核酸为基础的功能基因克隆和表达等分子实验提供了依据。To isolate high-quality extraction method for the genomic DNA from Paphiopedilum,genomic DNAs extraction from the mature leaves of five Paphiopedilum species(Paphiopedilum malipoense,Paphiopedilum purpuratum,Paphiopedilum dianthum,Paphiopedilum micranthum,and Paphiopedilum hirsutissimum) were studied using four methods,CTAB method,improved CTAB method,high salt low pH(HSLpH) method and SDS method in this paper.The integrity and purity of genomic DNA were detected by agarose gel electrophoresis and spectrophotometer.The results indicated that the modified CTAB method was suitable for high-quality genomic DNA extraction from all five species of Paphiopedilum and there were significant differences in the quality of the genomic DNA by the other three methods.The results suggested that CTAB method was suitable for high-quality genomic DNA extraction from P.purpuratum,P.dianthum and P.micranthum.SDS method was suitable for high-quality genomic DNA extraction from P.malipoense and P.hirsutissimum.Therefore,the extraction method obtaining of high-quality genomic DNA from Paphiopedilum laid the foundation for the subsequent molecular experiments such as the cloning and expression of functional genes.

关 键 词：兜兰 DNA提取 改良CTAB法 SDS法 

分 类 号：S682.31[农业科学—观赏园艺]