牡丹基因组AFLP银染反应体系的建立和优化

Establishment and Optimization of the AFLP Silver-staining Reaction System for Paeonia suffruticosa L.Genome

采用改进的SDS方法，从牡丹成熟叶片中提取基因组DNA，所得DNA样品的A260／A280值在1．7～1．8之间，琼脂糖凝胶电泳主带清晰、DNA纯度高、较少降解、不舍酶反应抑制剂，可被MseⅠ和PstⅠ两种限制性内切酶完全酶切．通过酶切连接、预扩增、选择性扩增、银染等试验过程，成功建立了牡丹品种AFLP银染反应体系。得到了清晰的指纹图谱，对扩增结果进行聚类分析，得到了23个牡丹品种的聚类分析图。

The improved method of SDS - DNA isolation was used to extract genome DNA from mature leaves of Paeonia suffruticosa L. High quality genome DNA was obtained. The values of A260/A280 of each sample were between 1.7 and 1.8. Strips in agarose electrophoresis were clear and less decomposed. These DNA samples with high purification were intact without containing inhibitor of enzyme reaction and could be completely digested by restriction enzymes such as Mse Ⅰ and Pst Ⅰ. The system suitable for silver-stained AFLP analysis were established successfully by processes like restriction enzymes digesting-ligasing reaction, preamplification,and selective amplification. Sharp fingerprintings were obtained by these processes. The dendrogram of 23 tree peony cuhivars was generated by clustering analysis of the amplified results.