栽培草莓叶绿体分离与cpDNA的提取方法  

An Efficient Protocol for Intact Chloroplast Isolation and cpDNA Extraction in Cultivated Strawberry

作　　者：程慧 陈宝玉 张燕[1] 陈莹[1] 乔玉山[1] CHENG Hui;CHEN Baoyu;ZHANG Yan;CHEN Ying;QIAO Yushan(College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China)

机构地区：[1]南京农业大学园艺学院,南京210095

出　　处：《西北植物学报》2017年第12期2496-2501,共6页Acta Botanica Boreali-Occidentalia Sinica

基　　金：江苏省农业科技自主创新资金(CX(15)1029)

摘　　要：以栽培草莓‘红颊’(Fragaria×ananassa Duch.‘Benihoppe’)新鲜幼嫩叶片为材料,利用高盐-低pH的缓冲液结合Percoll密度梯度离心分离纯化叶绿体,再用SDS-蛋白酶K法提取叶绿体DNA(cpDNA)。经荧光显微镜、琼脂糖凝胶电泳和PCR扩增检测,结果表明:该方法分离的草莓叶绿体完整性高,叶绿体DNA质量好,纯度高,能够满足后续基因组测序等实验的基本要求,为草莓属及其他草本植物cpDNA的提取提供参考。Using the fresh young leaves collected from cultivated strawberry(Fragaria×ananassa Duch.‘Benihoppe’)as material,we separated and purified the chloroplasts by high salt low pH buffer followed by Percoll density gradient centrifugation.The chloroplast DNA(cpDNA)was extracted by SDS Protein K method.The results of fluorescence microscope examination,agarose gel electrophoresis and PCR amplification showed that this method could obtain high integrity chloroplasts and high quality and purity cpDNA,which could meet the basic requirements of subsequent genome sequencing experiments.It could provide reference for extraction cpDNA of Fragaria and other herbaceous plants.

关 键 词：草莓 叶绿体分离 叶绿体DNA提取 高盐低pH Percoll密度梯度 

分 类 号：Q813[生物学—生物工程] Q503