川产贝母种质资源遗传多样性的ISSR分析

目的:为阐明川产贝母资源间亲缘关系提供更多证据.方法:利用ISSR分子标记技术对川产10个种1变种共19份材料进行分析,采用NTSYS软件计算材料间相似系数,并用UPGMA法构建了系统树.结果:从35个ISSR引物中共筛选出11个多态性明显、反应稳定的引物,在19份供试材料DNA中共扩增出179条谱带,其中多态性条带163条,占86.8%.材料间ISSR标记遗传相似性系数(GS)在0.569～0.855.聚类分析结果显示,所有供试材料均可区分开,并聚为4组.第1组包括川贝母、甘肃贝母、长腺贝母和短丝贝母,第Ⅱ组由暗紫贝母和浓蜜贝母组成,槽鳞贝母、浙贝母、瓦布贝母和梭砂贝母聚为第Ⅲ组,湖北贝母单独为第Ⅳ组.试验结果还表明,来源于同一地区的多数川产贝母材料可分别聚在一起.结论:ISSR标记适用于川产贝母资源的遗传多样性研究,但药典收载的川贝母药材的4种基原植物种间及其与其他川产贝母属各物种间尚不能仅通过采用ISSR标记技术进行分子鉴定.此外,ISSR聚类结果同川产贝母资源的地理分布有一定关系.

Genetic diversity of Fritillaria from Sichuan province based on ISSR

Objective: To provide more proofs for expounding the genetic relationships among the (varietal) species in genus Fritillaria from Sichuan province. Method: The ISSR marker technique was used to study relationships and genetic polymorphism of nineteen populations in ten species and one varietal species of genus Fritillaria. Genetic similarities were calculated by using NTSYS software and the dendrogram was constructed by using UPGMA method. Result: Eleven primers were selected from 35 ISSR primers, and 179 DNA fragments were amplified from 19 populations. Of which, 179 fragments were polymorphic (percentage of polymorphic bands was 86.8 %). The genetic similarity among all accessions ranged from 0.569 to 0.855. Clustering analysis showed that the 19 populations of Fritillaria could be distinctively classified into 4 groups. F. cirrhosa, F. przewalskii, F. cirrhosa var. longinectarea and F. dajinensis were in the first group; The second group was the cluster of F. cirrhosa and F. mellea (wild and cultivated species); The third group was F. sulcisquamosa, F. thunbergii, wabunesis and F. delavayi; F. hupehensis alone formed the fourth group. Conclusion: ISSR marker technique is suitable for the genetic diversity of Fritillaria from Sichuan province. Interspecific identifications among the four original species of Bulbus Fritillariae Cirrhosae recorded by pharmacopoeia of China, and between them and the other species of genus Fritillaria from Sichuan province could not be gained by using ISSR markers technique. In addition, the cluster result of genus Fritillaria had some relationships with the geographical distribution.