期 刊 ：植物研究 2015年 35卷 3期 页码：450-456
Keywords:Dipsacus plant, ISSR markers, genetic relatedness,
摘 要 ：续断属（Dipsacus L.）植物是中国续断科（Dipsacaceae）传统中药材，但属下种间亲缘关系仍存在不准确性鉴定。本研究拟采用10条多态性较高的ISSR（Inter-simple sequence repeat）标记从分子水平研究10份续断属植物的分类和种间亲缘关系，为该属植物的科学分类提供资料。从100条ISSR引物中筛选出多态性较高的10条，对10份续断材料进行PCR扩增，共扩增出947条DNA带，其中多态性条带828条，平均多态性条带百分率为87.4%，表明续断属植物具有丰富的遗传变异。遗传相似系数变化范围在0.557～0.806，平均为0.661。聚类分析结果表明，10份续断在相似系数水平0.582处可分为2个大类，进一步在0.702又可分为5个小类群。第1大类群包括3个小类群，第1小类群包括4分大理续断［DL(L),DL(W),DL(H),DL(WS)］，第2小类群包括日本续断（RB）和恩施续断(ES)，第3小类群包括2份川续断［C(J)和C(D)］，这表明小类群间品系亲缘关系比较近，基因交流比较频繁；第2大类群包括大头续断（DT）和深紫续断(SZ)，两者遗传距离较远，分别构成另外两个小类群。主成份分析与聚类分析所得的结果基本一致。研究结果还发现日本续断（RB）和恩施续断(ES)两者亲缘关系较近（0.210 1），且两者的地理分布有较大的重叠，可能两者基因交流比较频繁，由于环境的不同形态上稍有差异。推测恩施续断可能为日本续断的一个变种，当然这还需要进一步深入研究。
Abstract:Dipsacus L., a member of Dipsacaceae, is an important plant for the traditional herbal medicine in China. But there is still inaccurate identification in the botanical origin and genetic relatedness of Dipsacus plant. We used ten highly polymorphic ISSR(Inter-simple sequence repeat) markers to assess the genetic relatedness of ten accessions of Dipsacus, and used ten highly polymorphic ISSR primers from 100 primers screened to conduct PCR amplification with the genomic DNA isolated from ten accessions of Dipsacus. A total of 947 DNA bands were produced, of which 828 were polymorphic(87.2%) among ten accessions, and Dipsacus plant had abundant genetic variation. The Nei’s genetic coefficient ranged in 0.557 0.806 with an average of 0.661. By UPGMA cluster, ten Dipsacus accessions were grouped into two main groups(Group 1 and Group 2 ) with a similarity level of 0.582 and then into five subgroups at the similarity level of 0.702. Group 1 contained DL(L), DL(W), DL(H), DL(WS), ES, RB, C(J) and C(D) and was further divided into three subgroups(Subgroup 1, Subgroup 2 and Subgroup 3) and Group 2 consisted of DT and SZ, which was distance from any other accessions and constituted a separate minor subgroup(Subgroup 4 and Subgroup 5), respectively. Subgroup 1 contained DL(L), DL(W), DL(H) and DL(WS), Subgroup 2 with ES and RB and Subgroup 3 with C(J) and C(D), which indicated that the genetic relationship among them was very close and the gene flow among them was frequent. The results of principle coordinate analysis(PCA) were consistent with those of the UPGMA analysis. The distance between ES and RB was extremely narrow(0.210), which showed that RB and ES had a close relationship between them. The geographical distributions of them had overlapped intensively. The molecular cluster result corresponded to the morphology analysis. The results also showed that ES may be a variant of RB.