红豆杉AFLP分子标记技术体系的建立及应用

Establishment and Application of AFLP Molecular Marker Technology System of Taxus

采用改良的CTAB法提取红豆杉叶片的基因组DNA,得到适合于AFLP分析的DNA。通过对酶切、连接反应体系,预扩增和选择性扩增体系的调整;通过对聚丙烯酰胺凝胶电泳和银染方法的改进,建立了适合于红豆杉的AFLP分子标记体系;同时,应用该体系对红豆杉供试样品进行分析,发现不同种源的红豆杉之间存在多态性条带,证明红豆杉不同品系间存在差异。

The method of extracting genomic DNA from Taxus was investigated and the high quality genomic DNA was obtained.Several major factors affecting the results of AFLP fingerprints of Taxus were studied, including enzyme digestion and ligation,pre-amplification and selective amplification, polyacrylamide gel and silver staining, and an AFLP analysis system suitable for Taxus was established. By this system, the analysis of Taxus materials showed that there were polymorphic bands in different Taxus.