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目的:对形态上难以区分的湖北麦冬及其近缘植物,利用ISSR分子标记技术进行多样性研究,并探索其亲缘关系。方法:设计5因素4水平正交实验,进行PCR反应体系的优化;从ISSR引物序列表中随机选取80个引物,对所有样品进行PCR扩增,筛选实验有效引物;用优选的条件对样品进行ISSR分子标记,计算样品间的相似系数,构建聚类分析树系图。结果:建立了麦冬类植物优化的PCR扩增程序,筛选出了9个有效引物,得到了各样品的ISSR电泳谱和系统发育树系图,谱和图清楚反映出了各品种之间的区别及其亲缘关系。结论:本文建立的IS-SR分子标记技术可用于湖北麦冬及其近缘种的多样性及亲缘关系研究。
OBJECTIVE: To study the diversity of Ophiopogon japonicus and its related plants in Hubei Province, which are indistinguishable morphologically, using ISSR molecular markers technology and to explore their genetic relationship. Methods: Five factors and four orthogonal experiments were designed to optimize the PCR reaction system. Eighty random primers were selected from the sequence list of ISSR primers. All the samples were amplified by PCR, and the effective primers were screened. The samples were subjected to the optimal conditions ISSR molecular markers, calculate the similarity coefficient between samples and construct cluster analysis tree map. Results: The optimized PCR amplification program of Ophiopogon japonicus was established, 9 effective primers were screened, and the ISSR electrophoresis patterns and phylogenetic trees of each sample were obtained. The spectra and maps clearly showed that Differences and their relationship. Conclusion: The IS-SR molecular marker technology established in this paper can be used to study the diversity and genetic relationship of Ophiopogon japonicus and its related species in Hubei province.