白芷种质资源调查及其评价的研究

【作者】 郭丁丁；

【导师】 马逾英；

【作者基本信息】 成都中医药大学 ， 生药学， 2008， 硕士

【摘要】 为配合国家自然科学基金项目“白芷优良种质资源综合评价的研究”,本文在前期研究工作的基础上,对白芷种质资源进行了全面的调查、收集整理及评价研究,为白芷优良种质的筛选奠定了基础。首次对全国白芷种质资源进行了系统的调查、收集和整理,结合文献记载及全国白芷的商品情况,基本弄清了白芷种质资源的分布情况,了解了不同居群点的生态信息、栽培及生长特性、种植历史、栽种面积、生产现状、销售和使用情况等,同时采集原植物标本、收集种子及对口药材,并建立了白芷种质资源圃。在收集到的不同居群点白芷种子中选择主要居群点的样品进行种子的千粒种子重及水分测定试验,为白芷种子质量优劣的评价提供参考。按照《中国药典》2005年版白芷项下规定,对不同居群点白芷种质资源进行了主成分含量测定,为白芷优良种质资源的评价提供科学依据。结果显示:不同居群点白芷对口药材中醇溶性浸出物含量及欧前胡素含量存在明显差异;将不同居群点白芷栽种于四川遂宁白芷种质资源圃后,药材中浸出物含量及欧前胡素含量均有显著提高,反映了药材道地性对白芷内在品质的影响。在课题组前期工作的基础上,首次对不同居群点白芷对口药材及种质资源圃中的白芷药材进行了HPLC指纹图谱对比研究。通过对比图谱可以清楚地辨析不同居群点白芷样品中香豆素类成分的差异,同时进行了相似度和主成分的评价以及聚类分析。结果表明:兴安白芷与栽培白芷指纹图谱间的差异以及不同居群点栽培白芷指纹图谱间的差异主要体现在香豆素类成分各组分的比例和量上;将栽种到种质资源圃中的白芷药材进行相应的对比分析后发现,移栽后的白芷其化学表达随产地及栽培方法的一致而趋同,提示白芷中香豆素类成分与产地环境密切相关,产地生境或栽培技术可能是决定其质量的关键因素。本文首次将ISSR分子标记技术运用于白芷遗传多样性的研究,建立了重复性好、分辨率高、适用于白芷的ISSR反应体系。通过PCR扩增,共得到97条扩增条带,其中多态性条带37条,占38%,表明不同居群点白芷的遗传多样性水平较低,种质间的亲缘关系较近。依据基因遗传相似度将所有样品进行UPGMA聚类,栽培白芷样品的聚类没有表现出明显的规律性,不同居群点栽培白芷相互交叉的聚在一起形成一类而与兴安白芷相区别,该结果进一步证实了文献报道中认为川、杭、祁、禹四类传统商品白芷应属于同一类群的观点。更多还原

【Abstract】 In order to work for the project of the National Natural Science Foundation of China, this paper studies the distribution of resource, the ecological characters and the evaluation of germplasm of Baizhi on the basis of pre-effort of the project group,which supplys the evidence of studing and the evaluation germplasm resources.The germplasm resource of Baizhi has been investigated, collected and tidied methodically for the first time. We find out the Baizhi distribution in the producing areas of China and the information of their different habitats, the character of growing, the history of planting, the areas of planting and the situation of producing, selling and using. At the same time, we established the germplasm resource garden by investigating and collecting Baizhi in different areas.The Water and the thousand kernel weight of main populations have been determined, which have been provided quality evaluation about seed of Baizhi.The chief constituents of Baizhi have been detected by the relative methods of Chinese Pharmacopoeia(2005 edition), the results showed that there exist obvious differences on the content of the ethanol soluble extraction and the imperatorin, and the content of the ethanol soluble extraction and the imperatorin are significantly increased when the herbs are planted in the same place.The comparative study on Baizhi collected in different areas and planted in the germplasm resource garden has been done for the first time by the HPLC fingerprint. Drawing the conclusion: the environmental factors such as longitude, latitude, annual amount of precipitation all year around,etc.have great influences on the quality of Baizhi.ISSR is firstly used to analyze the level of the genetic diversity of Baizhi. Based on 9 primers, 97 ISSR bands are generated, 37(38%) are polymorphic, indicating a close relation genetic relationship. The result of the cluster analysis supports the viewpoint that the four cultivated breeds of Baizhi belong to the same taxon, which is different from the wild Baizhi.更多还原