天麻总DNA提取及聚合酶链反应扩增鉴定

The Isolation of Total DNA of Gastrodia.elata and Its Identification with PCR Amplification

目的:改良常用的植物DNA提取的CTAB,SDS法,以有效去除天麻多糖类杂质。方法:用CTAB法提取天麻鲜品不同部位(块茎、花序轴和胚芽)DNA;用生理盐水浸泡天麻干品之后,在应用CTAB,SDS法提取天麻干品的DNA;通过已知序列的天麻抗真菌蛋白基因片段的聚合酶链反应(PCR)扩增和PCR产物测序鉴定天麻DNA及其质量。结果:提取了44份天麻样本DNA,用CTAB法提取天麻鲜品不同部位的DNA,成功地用于PCR扩增,并通过DNA测序进一步鉴定;应用改良的CTAB,SDS法提取天麻干品DNA可用于PCR扩增,但DNA测序未成功。结论:用CTAB法提取的天麻鲜品各个部位DNA均可用于基因分析,而用天麻胚芽提取DNA的效果最理想;改良CTAB法能有效提取天麻干品DNA,提取的DNA可用于基因扩增。

Objective： To improve the common methods for extracting the plant DNA, so as to get rid of polyose impurity from Gastrodia elata frond. Methods： Using improved techniques of CTAB and SDS, we ideally extracted DNA from different parts of different samples of G. elata frond. Then, the extracted DNA was authenticated with PCR amplification and sequencing. Results： DNA extracted with CTAB technique from fresh samples of G. elata was successfully amplified with PCR technique and sequenced. DNA extracted with improved CTAB and SDS techniques from dried samples was also amplified with PCR, but its sequencing was not successful. Conclusions： DNA from all parts of G. elata fresh samples extracted with CTAB technique can be used for gene analysis, specially DNA from embryo bud. The improved CTAB technique can be used to extract DNA from dried samples of G. elata, but the DNA can be used only for gene amplification not for sequencing.