Study Of Hippophae On Qinghai-Tibet Plateau, Pharmacognosy And Genetic Relationship

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.