【作者】 肖蔚；

【导师】 张浩；

【作者基本信息】 四川大学 ， 生药学， 2007， 硕士

【摘要】 沙棘属(Hippophae)为胡颓子科(Elaeagnaceae)多年生雌雄异株的木本植物，具有耐寒、耐旱、耐贫瘠的特点，为广生态幅类型。我国是沙棘属植物种质资源最丰富的地区也是其起源中心、原始类群中心和类群分化中心。青藏高原是我国沙棘的集中分布地，具有巨大的资源蕴藏量而且是卧龙沙棘、棱果沙棘、理塘沙棘和密毛肋果沙棘等我国特有的珍稀类群的分布地区。本课题对青藏高原地区分布的沙棘属植物进行了资源调查；对10个不同基源的沙棘：中国沙棘(H．rhamnoides subsp．sinensis Rousi)、云南沙棘(H．rhamnoides subsp．yunnanensis Rousi．)、肋果沙棘(H．neurocarpa S．W．Liu et T．N．He)、密毛肋果沙棘(H．neurocarpa subsp．stellatopilosa Lian et al．ex Swenson et Bartish)、理塘沙棘(H．litangensis Lian et X．L．Chen ex Swenson et Bartish)、棱果沙棘(H．goniocarpa Lian et al．Swenson et Bartish)、西藏沙棘(H．tibetana Schlechtend．)、江孜沙棘(H．gyantsensis(Rousi)Lian)、卧龙沙棘(H．rhamnoides subsp．wolongensis Lian，K．Sun et X．L．Chen)和柳叶沙棘(H．Salicifolia D．Don)的分布地域、生境、原植物形态、叶子的显微特征进行了资源和生药学方面的调查研究。本课题应用ISSR—PCR技术对沙棘属植物(Hippophae L．)进行分析研究，探讨影响ISSR反应的各因子(模板DNA用量、TaqDNA聚合酶的用量、镁离子浓度、dNTP的浓度)，建立了沙棘的ISSR反应的优化反应体系，并应用于中国沙棘、云南沙棘、理塘沙棘、棱果沙棘、密毛肋果沙棘、肋果沙棘和西藏沙棘的亲缘关系进行研究。本课题对不同来源、不同采集地的沙棘叶样品中黄酮苷元的含量进行了测定和比较。并分别采用HPLC—UV和薄层色谱法，对10中不同来源的沙棘亚种进行分析，比较其沙棘叶中总黄酮组成和分布的相似性和差异性。更多还原

【Abstract】 The sea buckthorns are dioecious ligneous plants in the genus Hippophae, family Elaeagnaceae. They are naturally distributed in China and Europe with highly ecological, environmental and economical values that are contributed by their distinctive resistance to extreme climax and poor soil conditions. Therefore, they are able to survive and flourish in different growing places characterised by relatively various ecology types. China is where the most species and sub-species of natural sea buckthorn resources distribute. Also, it is the location where this genus had arisen and developed. In China, the sea buckthorn resources primarily distribute on Qinghai-Tibet Plateau. Furthermore, it is the only area that some rare subspecies exist, such as H. neurocarpa S. W. Liu et T. N. He, H. neurocarpa subsp. stellatopilosa Lian et al.ex Swenson et Bartish, H. litangensis Lian et X. L. Chen ex Swenson et Bartish, and H. goniocarpa Lian et al. Swenson et Bartish.In this project, firstly, the resource distribution of sea buckthorns was investigated and recorded, this study included all the 10 species or subspecies on Qinghai-Tibet Plateau, which are H. rhamnoides subsp. sinensis Rousi, H. rhamnoides subsp. yunnanensis Rousi, H. neurocarpa S. W. Liu et T. N. He, H. neurocarpa subsp. stellatopilosa Lian et al.ex Swenson et Bartish, H. litangensis Lian et X. L. Chen ex Swenson et Bartish, H. goniocarpa Lian et al. Swenson et Bartish, H. tibetana Schlechtend., H. gyantsensis (Rousi) Lian, H. rhamnoides subsp. wolongensis Lian, K. Sun et X. L. Chen) and H. salicifolia D.Don. The research work of living environments, botanical origins, berry and leaf’s morphological and microscopic characteristics were encompassed as well.Secondly, molecular biology method ISSR-PCR was established. The influential factors of ISSR—PCR amplification, such as template DNA usage, Taq DNA polymerase usage, Mg2+ concentration, dNTP concentration, were optimized and determined when they were applied to the sea buckthorn ISSR-PCR amplification system. The result further served the study on relationships of 7 subspecies and species of sea buckthorns. In this section, 12 primers were screened out from 100 random primers published by Colombia University, and a total of 86 DNA fragments were amplified. The clustering analysis and genetic distances were calculated by NTS YS software. The result of PCR reaction was not completely consistent with that of the morphological classification.The total flavonoids contents(TFC) of sea buckthorn leaves were studied and discussed by content determination and components comparison of TFC, the former experiment was on the basis of HPLC, and the latter part employed both HPLC and TLC. The leaves of all the 10 sea buckthorns of different origins were experimental samples. In this sense, the established methods could serve as preliminary chemotaxonomy evidence for species classification, relationship location as well as for helping understand the total flavonoids’ qualitative and quantitative characteristics such as their chemical constitution and components variation in different subspecies.更多还原