茶树叶绿体DNA提取方法研究

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

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.