茶树叶绿体DNA提取方法研究  

Extraction Method of Chloroplast DNA of Tea Plant(Camellia sinensis)

作　　者：陈春梅[1] 陈亮[1] 

机构地区：[1]中国农业科学院茶叶研究所、国家茶树改良中心、农业部茶树生物学与资源利用重点实验室,杭州310008

出　　处：《分子植物育种》2014年第3期562-566,共5页Molecular Plant Breeding

基　　金：国家茶叶产业技术体系项目(CARS-23);国家自然科学基金项目(30901159;31170624;31100504)共同资助

摘　　要：为了探究茶树叶绿体DNA（chloroplast DNA，cpDNA）的提取方法，本研究采用改进的高盐-低pH法和Percoll密度梯度法提取cpDNA。结果表明，两种方法提取的cpDNA经分光光度计检测，A260/A280值在1．80～1．90，高盐-低pH法提取的cpDNA产率为0．65μg/g，Percoll密度梯度法提取的cpDNA产率为0．07μg/g。提取的cpDNA经PCR检测，均能扩增出目标片段，但高盐-低pH法效果更好。研究结果为进一步研究茶树叶绿体基因组结构、功能等方面奠定基础。To explore an efficient method of extracting chloroplast DNA （cpDNA） from tea plant, the modified High-salt Low-pH and Percoll Density Gradient Centrifugation methods were compared and evaluated. The results showed that both methods successfully isolating good-quality cpDNA with an A260/280 ratio between 1.80-1.90, however, a higher yield was obtained by the modified High-salt Low-pH method （0.65 μg/g） as compared to the other method （0.07 μg/g）. In addition, although cpDNA extracted using both methods could serve as a template for PCR, better results were obtain with the modified High-salt Low-pH method. This research lays the groundwork for sequencing and analyzing of the entire chloroplast genome of tea plant.

关 键 词：茶树(Camellia sinensis) 叶绿体DNA 高盐-低pH法 密度梯度法 

分 类 号：S571.1[农业科学—茶叶生产加工]