甘肃祁连山地区麻花秦艽UPLC指纹图谱与遗传多样性研究

UPLC Fingerprints and Genetic Diversity of Gentiana straminea Maximfile Qilian Mountain Areas of Gansu Province

目的建立麻花秦艽UPLC指纹图谱,并分析其指征性成分差异与遗传多样性。方法应用超高效液相色谱法测定采集样本干燥根的主要化学成分,利用中药色谱指纹图谱相似度评价系统分析不同居群指征性成分共有峰的相对峰面积,应用SPSS19.0对其进行聚类分析;应用ISSR分子标记对所采集样本干燥叶片进行基因组DNA提取和扩增,利用Popgen32软件分析遗传多样性及遗传分化状况。结果 18个居群麻花秦艽药材共标定了12个共有峰,可分为3个类群,张掖市山丹县麻花秦艽龙胆苦苷及5种化学成分总含量最高;6条ISSR引物共扩增出73个位点,PPB为98.63%,I平均值为0.366 3,H平均值为0.231 6,Gst为0.344 1,基因流Nm为0.953 0。结论建立的麻花秦艽UPLC指纹图谱可作为其质量评价体系;麻花秦艽在种群水平上具有较高的遗传多样性,居群内遗传多样性水平明显高于居群间,遗传变异主要存在于居群内。

OBJECTIVE To establish the UPLC fingerprints of G. stramirtea , and analyze its differences of indications ingredient and genetic diversity. METHODS The main chemical composition of dry root were measured by UPLC. Similarity evaluation system for chromatographic fingerprint of TCM（Version 2004 A） was used to analyze the relative peak area of common peaks from differently populations ,and cluster analysis of common peaks was carried out by SPSS19. 0. Genomic DNA of dry leaves collected were extracted and amplified by ISSR markers. The genetic diversity and genetic distance were calculated by Popgen 32. RESULTS Twelve Com- mon peaks were selected as the fingerprint peaks from 18 populations of G. straminea, and three groups were divided according to the clustering results of chemical composition. It had the highest levels that was gentiopicroside content and total content of five kinds of chemical components in Shandan County, Zhangye City. Six primers produced 73 bands, among which 72 were polymorphic bands, the average percentage of polymorphie bands （PPB） was 98. 63 % ; Shannon＇ s information index （ I ） was 0. 366 3 ; Nei＇s genetic diversity index（H） was 0. 231 6. The genetic differentiation coefficient （Gst） and gene flow（Nm） of wild populations were 0. 344 1 and 0. 953 0. CONCLUSION UPLC Fingerprint of G. straminea can be used as the quality evaluation system. The genetic diversity among species of G. straminea is at higher level, but the level of genetic diversity within populations is higher than that between populations, and genetic variance mainly existed among populations.