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资助项目
GST, p < 0.01). At the regional level, Chinese and Japanese L. hodgsonii had a similar estimate of genetic diversity (China: Hd = 0.847, HT = 0.869; Japan: Hd = 0.766, HT = 0.867). Populations from China and Japan possess unique sets of haplotypes, and no haplotypes were shared between the regions. Furthermore, both the phyloegenetic and network analyses recovered the haplotypes of China and Japan as two distinct clades. Thus, we suggested the disjunct distribution of L. hodgsonii in China and Japan may present the climatic vicariant relicts of the ancient widely distributed populations. After divergence, this species within each region experienced independent evolutionary process. In China, L. hodgsonii was distributed around the Sichuan Basin. This distribution range can be divided into five regions. They were Jiajin Mountain region, E’mei Mountain region, Yunnan-Guizhou Plateau region, Wushan-Wuling Mountain region and Qinling Mountain region. Twelve haplotypes were indentified within these regions. Each region had its own specific haplotypes, which had different ancestry in the network. We deduced that Chinese L. hodgsonii might survive the LGM in multiple isolated refugia around the Sichuan Basin. In Japan, L. hodgsonii was disjunctively distributed in northern Honshu and Hokkaido. Seven haplotypes were identified within this region. However, the genetic diversity in Honshu (Hd = 0.821) was much higher than that in Hokkaido (Hd = 0.513). And all haplotypes in Hokkaido were derived from Honshu. This haplotype distribution suggested that the northern Honshu could have served as refuge in Japan. Nested clade analysis (NCA) indicated multiple forces including the vicariance and long-distance dispersal affected the disjunctive distribution among populations of L. hodgsonii in Japan.2. The phylogeography of L. tongolensis，Ligularia tongolensis was distributed along the Jinshajiang watershed, Yalongjiang watershed and Wumeng Mountain. In order to deduce the demographic history of this species, we sequenced two chloroplast DNA (cpDNA) intergenic spacers (trnQ-5’rps16, trnL-rpl32) in 140 individuals from 14 populations of three groups (Jinshajiang vs. Yalongjiang vs. Wumeng) within this species range. High levels of haplotype diversity (Hd = 0.814) and total genetic diversity (HT = 0.862) were detected at the species level, based on a total oftwelve haplotypes identified. However, the intrapopulation diversity (HS = 0.349) was low, which led to the high levels of genetic divergence (GST = 0.595, NST = 0.614, FST = 0.597). In consideration of the speciation of L. tongolensis resulting from the uplifts of the Qinghai-Tibetan Plateau (QTP), we thought the present genetic structure of L. tongolensis was shaped by the fragmentation of ancestral populations during the courses of QTP uplifts. This was further supported by the absence of IBD tests (r = –0.291, p = 0.964), which suggest that the differentiation had not occurred in accordance with the isolation by distance model. The genetic differentiation in L. tongolensis appears to be associated with historical events. Meanwhile, H2 and H5, the dominant haplotypes that located on internal nodes and deviated from extinct ancestral haplotype in the network, were detected to be shared between Jinshajiang and Yalongjiang groups. We deduced that ancestral populations of this species might have had a continuous distribution range, which was then fragmented and isolated by the following tectonic events. Finally, the ancestral polymorphism, H2 and H5, were randomly allocated in Jinshajiang watershed and Yalongjiang watershed. Meanwhile, H5 was the dominant haplotype in Jinshajiang watershed; H7 was the domiant haplotype in Yalongjiang watershed and Wumeng Mountain. This haplotype distribution pattern indicated that each group might have served as a refuge for L. tongolensis during the Quaternary Glaciation. Postglacial demographic expansion was supported by unimodal mismatch distribution and star-like phylogenies, with expansion ages of 274 ka B. P. for this 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search for new and efficient antitumor drugs from natural products, especially plants, has been the focus of scholars and researchers all over the world. The dissertation composed of four chapters and focused on antitumor drugs of plant origin and plant cyclopeptides research. The first chapter was about the establishment and application of cancer cell line assay. In total, 8835 compounds and 3638 extracts were tested against seven cell lines and the results indicated that 459 compounds and 189 extracts showed cytotoxicities. Based on these results, the plant Rubia yunnanensis (Franch.) Diels, which showed potential activity and has been used as a common herb to treat cancer, was selected for the further study. Chapter two, focused on the studies on chemistry, bioactivities, antitumor mechanism and preliminary pre-clinical research of cyclopeptides in R. yunnanensis. A phytochemical investigation on cyclopeptides constituents from this plant led to the isolation of 18 cyclopeptides, including 2 new ones with novel skeletons and 7 other new ones. The possible biosynthetic pathways for four types of rubiaceae-type cyclopeptides were also proposed. The 18 cyclopeptides were evaluated for their cyctotoxicities against 11 cancer cell lines and the result indicated that these cyclopeptides exhibited strong activities and RA-V showed best activities with IC50 values of approximately 10 nM. Moreover, RA-V was also found to exhibit anti I-type herpes simplex virus (HSV-1) activity for the first time. Thereafter, we carried out research on the antitumor mechanism of RA-V and its glycoside RA-XII. The results showed that they significantly inhibited TNF-α induced NF-κB signaling pathway. The phosphorylation of IκBα was down-regulated by them, which could inhibit the translocation of P65 and NO production in LPS and IFN-γ-induced RAW 264.7 murine macrophages. This is the first time it is being reported that RA-V and RA-XII are new natural NF-κB pathway inhibitors. Furthermore, from the anti-angiogenesis study, RA-V showed strong anti-proliferative activities in human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMEC). Further study on its mechanism indicated that RA-V could induce G2/M phase arrest and cell apoptosis by down-regulation of cell cycle regulatory protein and gene expression in extracellular signal-regulated kinase ½ (Erk ½) phosphorylation pathway. Moreover, RA-V could inhibit migration and tube formation of HUVEC and HMEC by inhibition of matrix metalloproteinase. The third chapter was about the studies on other chemical constituents of R. yunnanensis and their bioactivities. A phytochemical assessment of this plant resulted in the isolation of 68 compounds, including 11 new arborinane-type triterpenoids and 4 new quinones. Subsequently, 61 of them were evaluated for their cytotoxicities against three cancer cell lines, anti-Staphylococcus aureus and anti-Candida albicans activities. The results indicated that for the first time the arborinane-type triterpenoids exhibited cytotoxicities and also rubiarbonol G and 2-methyl-1,3,6-trihydroxy-9,10-anthraquinone induced cell apoptosis in Hela cell line. The fourth chapter was a review on chemical constituents and bioactivities of genus Rubia plants. All the above-mentioned research work had laid a good foundation for quality control and multi-component, multi-target drug R&D of R. yunnanensis.","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.source_filter&advanced=false&query1=%25E6%25A3%2580%25E6%25B5%258B%25E6%2596%25B9%25E6%25B3%2595&&fq=dc.project.title_filter%3AThe%5C+search%5C+for%5C+new%5C+and%5C+efficient%5C+antitumor%5C+drugs%5C+from%5C+natural%5C+products%2C%5C+especially%5C+plants%2C%5C+has%5C+been%5C+the%5C+focus%5C+of%5C+scholars%5C+and%5C+researchers%5C+all%5C+over%5C+the%5C+world.%5C+The%5C+dissertation%5C+composed%5C+of%5C+four%5C+chapters%5C+and%5C+focused%5C+on%5C+antitumor%5C+drugs%5C+of%5C+plant%5C+origin%5C+and%5C+plant%5C+cyclopeptides%5C+research.%5C+The%5C+first%5C+chapter%5C+was%5C+about%5C+the%5C+establishment%5C+and%5C+application%5C+of%5C+cancer%5C+cell%5C+line%5C+assay.%5C+In%5C+total%2C%5C+8835%5C+compounds%5C+and%5C+3638%5C+extracts%5C+were%5C+tested%5C+against%5C+seven%5C+cell%5C+lines%5C+and%5C+the%5C+results%5C+indicated%5C+that%5C+459%5C+compounds%5C+and%5C+189%5C+extracts%5C+showed%5C+cytotoxicities.%5C+Based%5C+on%5C+these%5C+results%2C%5C+the%5C+plant%5C+Rubia%5C+yunnanensis%5C+%5C%28Franch.%5C%29%5C+Diels%2C%5C+which%5C+showed%5C+potential%5C+activity%5C+and%5C+has%5C+been%5C+used%5C+as%5C+a%5C+common%5C+herb%5C+to%5C+treat%5C+cancer%2C%5C+was%5C+selected%5C+for%5C+the%5C+further%5C+study.%5C+Chapter%5C+two%2C%5C+focused%5C+on%5C+the%5C+studies%5C+on%5C+chemistry%2C%5C+bioactivities%2C%5C+antitumor%5C+mechanism%5C+and%5C+preliminary%5C+pre%5C-clinical%5C+research%5C+of%5C+cyclopeptides%5C+in%5C+R.%5C+yunnanensis.%5C+A%5C+phytochemical%5C+investigation%5C+on%5C+cyclopeptides%5C+constituents%5C+from%5C+this%5C+plant%5C+led%5C+to%5C+the%5C+isolation%5C+of%5C+18%5C+cyclopeptides%2C%5C+including%5C+2%5C+new%5C+ones%5C+with%5C+novel%5C+skeletons%5C+and%5C+7%5C+other%5C+new%5C+ones.%5C+The%5C+possible%5C+biosynthetic%5C+pathways%5C+for%5C+four%5C+types%5C+of%5C+rubiaceae%5C-type%5C+cyclopeptides%5C+were%5C+also%5C+proposed.%5C+The%5C+18%5C+cyclopeptides%5C+were%5C+evaluated%5C+for%5C+their%5C+cyctotoxicities%5C+against%5C+11%5C+cancer%5C+cell%5C+lines%5C+and%5C+the%5C+result%5C+indicated%5C+that%5C+these%5C+cyclopeptides%5C+exhibited%5C+strong%5C+activities%5C+and%5C+RA%5C-V%5C+showed%5C+best%5C+activities%5C+with%5C+IC50%5C+values%5C+of%5C+approximately%5C+10%5C+nM.%5C+Moreover%2C%5C+RA%5C-V%5C+was%5C+also%5C+found%5C+to%5C+exhibit%5C+anti%5C+I%5C-type%5C+herpes%5C+simplex%5C+virus%5C+%5C%28HSV%5C-1%5C%29%5C+activity%5C+for%5C+the%5C+first%5C+time.%5C+Thereafter%2C%5C+we%5C+carried%5C+out%5C+research%5C+on%5C+the%5C+antitumor%5C+mechanism%5C+of%5C+RA%5C-V%5C+and%5C+its%5C+glycoside%5C+RA%5C-XII.%5C+The%5C+results%5C+showed%5C+that%5C+they%5C+significantly%5C+inhibited%5C+TNF%5C-%CE%B1%5C+induced%5C+NF%5C-%CE%BAB%5C+signaling%5C+pathway.%5C+The%5C+phosphorylation%5C+of%5C+I%CE%BAB%CE%B1%5C+was%5C+down%5C-regulated%5C+by%5C+them%2C%5C+which%5C+could%5C+inhibit%5C+the%5C+translocation%5C+of%5C+P65%5C+and%5C+NO%5C+production%5C+in%5C+LPS%5C+and%5C+IFN%5C-%CE%B3%5C-induced%5C+RAW%5C+264.7%5C+murine%5C+macrophages.%5C+This%5C+is%5C+the%5C+first%5C+time%5C+it%5C+is%5C+being%5C+reported%5C+that%5C+RA%5C-V%5C+and%5C+RA%5C-XII%5C+are%5C+new%5C+natural%5C+NF%5C-%CE%BAB%5C+pathway%5C+inhibitors.%5C+Furthermore%2C%5C+from%5C+the%5C+anti%5C-angiogenesis%5C+study%2C%5C+RA%5C-V%5C+showed%5C+strong%5C+anti%5C-proliferative%5C+activities%5C+in%5C+human%5C+umbilical%5C+vein%5C+endothelial%5C+cells%5C+%5C%28HUVEC%5C%29%5C+and%5C+human%5C+microvascular%5C+endothelial%5C+cells%5C+%5C%28HMEC%5C%29.%5C+Further%5C+study%5C+on%5C+its%5C+mechanism%5C+indicated%5C+that%5C+RA%5C-V%5C+could%5C+induce%5C+G2%5C%2FM%5C+phase%5C+arrest%5C+and%5C+cell%5C+apoptosis%5C+by%5C+down%5C-regulation%5C+of%5C+cell%5C+cycle%5C+regulatory%5C+protein%5C+and%5C+gene%5C+expression%5C+in%5C+extracellular%5C+signal%5C-regulated%5C+kinase%5C+%C2%BD%5C+%5C%28Erk%5C+%C2%BD%5C%29%5C+phosphorylation%5C+pathway.%5C+Moreover%2C%5C+RA%5C-V%5C+could%5C+inhibit%5C+migration%5C+and%5C+tube%5C+formation%5C+of%5C+HUVEC%5C+and%5C+HMEC%5C+by%5C+inhibition%5C+of%5C+matrix%5C+metalloproteinase.%5C+The%5C+third%5C+chapter%5C+was%5C+about%5C+the%5C+studies%5C+on%5C+other%5C+chemical%5C+constituents%5C+of%5C+R.%5C+yunnanensis%5C+and%5C+their%5C+bioactivities.%5C+A%5C+phytochemical%5C+assessment%5C+of%5C+this%5C+plant%5C+resulted%5C+in%5C+the%5C+isolation%5C+of%5C+68%5C+compounds%2C%5C+including%5C+11%5C+new%5C+arborinane%5C-type%5C+triterpenoids%5C+and%5C+4%5C+new%5C+quinones.%5C+Subsequently%2C%5C+61%5C+of%5C+them%5C+were%5C+evaluated%5C+for%5C+their%5C+cytotoxicities%5C+against%5C+three%5C+cancer%5C+cell%5C+lines%2C%5C+anti%5C-Staphylococcus%5C+aureus%5C+and%5C+anti%5C-Candida%5C+albicans%5C+activities.%5C+The%5C+results%5C+indicated%5C+that%5C+for%5C+the%5C+first%5C+time%5C+the%5C+arborinane%5C-type%5C+triterpenoids%5C+exhibited%5C+cytotoxicities%5C+and%5C+also%5C+rubiarbonol%5C+G%5C+and%5C+2%5C-methyl%5C-1%2C3%2C6%5C-trihydroxy%5C-9%2C10%5C-anthraquinone%5C+induced%5C+cell%5C+apoptosis%5C+in%5C+Hela%5C+cell%5C+line.%5C+The%5C+fourth%5C+chapter%5C+was%5C+a%5C+review%5C+on%5C+chemical%5C+constituents%5C+and%5C+bioactivities%5C+of%5C+genus%5C+Rubia%5C+plants.%5C+All%5C+the%5C+above%5C-mentioned%5C+research%5C+work%5C+had%5C+laid%5C+a%5C+good%5C+foundation%5C+for%5C+quality%5C+control%5C+and%5C+multi%5C-component%2C%5C+multi%5C-target%5C+drug%5C+R%5C%26D%5C+of%5C+R.%5C+yunnanensis."},{"jsname":"Transposable elements (TEs) have been found to be a significant fraction of eukaryotic genomes. Moreover, they make great contributions to the structure, function and evolution of genomes as well as genes. However, some questions such as the mechanisms of retainment of TEs in the genome and their adaptive evolution have not been fully elucidated so far. In this study, the distributions of 17 TE-gene associations among Oryza species were investigated. In addition, the nucleotide diversity was analysed and neutral tests for the region flanking the TE insertions were performed. Based on the above-observed patterns, evolutionary relationships between species in the AA genome group were discussed. The main results are as follows: For each TE-gene association, PCR and electrophoresis were conducted for a total of 107 strains, belonging to different Oryza species. The patterns of each TE-gene association in different species were obtained. It is our finding that 2 associations distribute through all Oryza species. By contrast, other 15 associations were only observed in some Oryza species. On basis of the above-mentioned results, it is likely that insertion events under study occurred in their common ancestor, and then they dispersed with subsequent divergence of different AA genome species. Our datas strongly support that O. meridionalis is the most basal lineage of AA genome group, instead of O. longistaminata.For several TE-gene associations fixed in populations of ancestor, the nucleotide diversity was estimated and neutral tests for the region flanking the TE insertions between populations with and without TE insertions were performed. No significant result was obtained. It is possible that the fixation of mutations with TE insetion is a random process; alternatively, this process is attributable to nature selection. Since the fixation has finished, it is difficult to detect the signature at the sequence level.","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.source_filter&advanced=false&query1=%25E6%25A3%2580%25E6%25B5%258B%25E6%2596%25B9%25E6%25B3%2595&&fq=dc.project.title_filter%3ATransposable%5C+elements%5C+%5C%28TEs%5C%29%5C+have%5C+been%5C+found%5C+to%5C+be%5C+a%5C+significant%5C+fraction%5C+of%5C+eukaryotic%5C+genomes.%5C+Moreover%2C%5C+they%5C+make%5C+great%5C+contributions%5C+to%5C+the%5C+structure%2C%5C+function%5C+and%5C+evolution%5C+of%5C+genomes%5C+as%5C+well%5C+as%5C+genes.%5C+However%2C%5C+some%5C+questions%5C+such%5C+as%5C+the%5C+mechanisms%5C+of%5C+retainment%5C+of%5C+TEs%5C+in%5C+the%5C+genome%5C+and%5C+their%5C+adaptive%5C+evolution%5C+have%5C+not%5C+been%5C+fully%5C+elucidated%5C+so%5C+far.%5C+In%5C+this%5C+study%2C%5C+the%5C+distributions%5C+of%5C+17%5C+TE%5C-gene%5C+associations%5C+among%5C+Oryza%5C+species%5C+were%5C+investigated.%5C+In%5C+addition%2C%5C+the%5C+nucleotide%5C+diversity%5C+was%5C+analysed%5C+and%5C+neutral%5C+tests%5C+for%5C+the%5C+region%5C+flanking%5C+the%5C+TE%5C+insertions%5C+were%5C+performed.%5C+Based%5C+on%5C+the%5C+above%5C-observed%5C+patterns%2C%5C+evolutionary%5C+relationships%5C+between%5C+species%5C+in%5C+the%5C+AA%5C+genome%5C+group%5C+were%5C+discussed.%5C+The%5C+main%5C+results%5C+are%5C+as%5C+follows%5C%3A%5C+For%5C+each%5C+TE%5C-gene%5C+association%2C%5C+PCR%5C+and%5C+electrophoresis%5C+were%5C+conducted%5C+for%5C+a%5C+total%5C+of%5C+107%5C+strains%2C%5C+belonging%5C+to%5C+different%5C+Oryza%5C+species.%5C+The%5C+patterns%5C+of%5C+each%5C+TE%5C-gene%5C+association%5C+in%5C+different%5C+species%5C+were%5C+obtained.%5C+It%5C+is%5C+our%5C+finding%5C+that%5C+2%5C+associations%5C+distribute%5C+through%5C+all%5C+Oryza%5C+species.%5C+By%5C+contrast%2C%5C+other%5C+15%5C+associations%5C+were%5C+only%5C+observed%5C+in%5C+some%5C+Oryza%5C+species.%5C+On%5C+basis%5C+of%5C+the%5C+above%5C-mentioned%5C+results%2C%5C+it%5C+is%5C+likely%5C+that%5C+insertion%5C+events%5C+under%5C+study%5C+occurred%5C+in%5C+their%5C+common%5C+ancestor%2C%5C+and%5C+then%5C+they%5C+dispersed%5C+with%5C+subsequent%5C+divergence%5C+of%5C+different%5C+AA%5C+genome%5C+species.%5C+Our%5C+datas%5C+strongly%5C+support%5C+that%5C+O.%5C+meridionalis%5C+is%5C+the%5C+most%5C+basal%5C+lineage%5C+of%5C+AA%5C+genome%5C+group%2C%5C+instead%5C+of%5C+O.%5C+longistaminata.For%5C+several%5C+TE%5C-gene%5C+associations%5C+fixed%5C+in%5C+populations%5C+of%5C+ancestor%2C%5C+the%5C+nucleotide%5C+diversity%5C+was%5C+estimated%5C+and%5C+neutral%5C+tests%5C+for%5C+the%5C+region%5C+flanking%5C+the%5C+TE%5C+insertions%5C+between%5C+populations%5C+with%5C+and%5C+without%5C+TE%5C+insertions%5C+were%5C+performed.%5C+No%5C+significant%5C+result%5C+was%5C+obtained.%5C+It%5C+is%5C+possible%5C+that%5C+the%5C+fixation%5C+of%5C+mutations%5C+with%5C+TE%5C+insetion%5C+is%5C+a%5C+random%5C+process%5C%3B%5C+alternatively%2C%5C+this%5C+process%5C+is%5C+attributable%5C+to%5C+nature%5C+selection.%5C+Since%5C+the%5C+fixation%5C+has%5C+finished%2C%5C+it%5C+is%5C+difficult%5C+to%5C+detect%5C+the%5C+signature%5C+at%5C+the%5C+sequence%5C+level."},{"jsname":"Tupistra pingbianensis J. L. Huang & X. Z. Liu, is a newly described perennial herb narrowly distributed in South-east Yunnan, China. It belongs to genera Tupistra Ker Gawler(Liliaceae). It usually occurs on outcrops of bare rock, or occasionally as an epiphyte on tree trunks covered with humus and moss. T. pingbianensis is unusual in that it exhibits rarity according to three different ways of measuring rarity, i.e. it has a small geographical range, is a habitat specialist, and always has low abundance where it occurs. Because of this, T. pingbianensis has been listed as an endangered species and catalogued in the Chinese Species Red List. In order to discuss the causes of rarity of T. pingbianensis, the multidisciplinary investigations of the seed and seedling establishment, cytology, breeding system, and population genetic structure of the endangered T. pingbianensis were performed in this thesis. Besides, the corresponding conservation strategies were also proposed according to the above-mentioned. The main results are summarized as follows:1. Biological traits of T. pingbianensis,T. pingbianensis is a perennial herbaceous with a creeping rhizome, thick basal leaves, and an inflorescence that is a terminal spike. Florescence is from November to December, while fruiting occurs between November and December in the next year. Reproduction and spread also occurs clonally via rhizomes, most seeds simply fall from the mother plant and germinate where they land. It occurs on outcrops of bare rock, or occasionally as an epiphyte on tree trunks covered with humus and moss, which are naturally rare habitat. Throughout its small geographical range, T. pingbianensis occurs as discrete, small populations size. 2. Seed germination traits of T. pingbianensis,Seed morphology was observed and effects of substrates soil types, light, sowing depth on germination percentage of the species T. pingbianensis were investigated primarily. The results showed that the average seed size was (1.17±0.02) cm × (0.79±0.01) cm × (0.77±0.01) cm (length × width × thickness), per-hundred-seed-weight was about 35.03±0.12g. Among the three different substrates soil types and sowing depths, seeds of T. pingbianensis germinate best in alkalescence soil and shallow sowing depth (2cm). It could germinate in the both light and dark, but the germination rate can be accelerated by light obviously. Its seed has high germination rate not just in greenhouse, but also in the field. We considered that this is a good strategy to expand its population in the special habit.3. Karyotype evolution status of T. pingbianensis,The karyotype of total eight species in Campylandra, Tupistra and Aspidistra from China were reported. Considering Tupistra has the similar morphological character with Campylandra but resemble Aspidistra in karyotype. The results support the earlier study that Tupistra is a transition between Compylandra and Aspidistra. Besides, our results also showes that the T. pingbianensis and T. fungilliformis has higher karyotype asymmetry than other species in this genera, which means these species have higher karyotype evolution status. 4. Reproduction ecology of T. pingbianensis, The flower phenology, pollinators of T. pingbianensis were documented herein. We also examined the breeding system of T. pingbianensis and seed fitness traits to determine what forms of pollination and mating occur in this naturally rare species, and is there evidence of inbreeding depression in its populations. The results shows that the flowers opened 10-15 days, which suggest stigma and pollen can keep high vitality for a long time (10-15 days). The only pollinators observed on T. pingbianensis flowers were ants (Aphaenogaster smythiesii Forel,Formicidea), springtail (Hypogastrura sp., Hypogastruridae, Collembola) and one species of beetles (Anomala corpulenta Motsch, Rutelidae). These pollinators generally have restricted movement capacities and hence promote geitonogamy or mating between individuals in close proximity within populations. The results of out crossing index (OCI) pollination experiments in our study suggest that T. pingbianensis has an animal-pollinated, mixed selfing and outcrossing breeding systems. However, a pollination experiment also fail to detect significant inbreeding depression upon F1 fruit set, seed weight and germinate rate fitness-traits. Since naturally rare species T. pingbianensis is not seriously genetically impoverished and likely to have adapted to tolerating a high level of inbreeding early in its history. 5. Conservation genetic of T. pingbianensis, The levels and partitioning of genetic diversity were investigated in Tupistra pingbianensis. Here genetic diversity and patterns of genetic variation within and among 11 populations were analyzed using AFLP markers with 97 individuals across its whole geographical range. High levels of genetic variation were revealed both at the species level (P99 = 96.012%; Ht = 0.302) and at the population level (P99 = 51.41%; Hs = 0.224). Strong genetic differentiation among populations was also detected (FST = 0.2961; ⍬Ⅱ= 0.281), which corresponded to results reported for typical animal-pollinated, mixed selfing and outcrossing plant species. Special habitat and its life history traits may play an important role in shaping the genetic diversity and the genetic structure of this species. Based on the special habitat in T. pingbianensis, the most suitable strategy for its conservation is the protection of its habitat. Moreover, given the observed strong genetic differentiation among populations of T. pingbianensis, the preservation of genetic diversity in this species will require the protection of many populations as possible to maintain the current levels of genetic 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a town of Shangri-la County, Diqing Prefecture, was chosen as the main field site for studying the structure and characters of traditional agricultural ecosystem, by using approaches of ethnobotany, cultural anthropology and ecology. Combined with interviewing exercises in Hanpi village, Jiantang Township, this paper also discussed the impact of traditional management on the biocultural diversity. The results showed: Traditional agroecosystem in Shangri-la is an integrated system with three subsystems, which are farming, forest and grazing subsystem. The seasonal shifting grazing activity in Shangri-la, following the natural season change and the recover process of plants, is a sustainable management that protects the local biodiversity. However, along with the decay of shifting grazing tradition recently, the local Tibetans turned to use grass land and forest which is close to villages as the main grazing lands. It increased the pasturing pressure to these areas and caused productivity decreasing and biodiversity. As a symbolic part of Tibetan culture in Shangri-la, the sacred mountain culture has played a significant role in biodiversity conservation by restricting human’s behavior. The Tibetan traditional culture, indigenous knowledge and traditional ecosystem management in Shangri-la has contributed to the biodiversity conservation in this area. However, this research indicated that under the pressure of mainstream culture and market economy, traditional knowledge is vanishing; old crop land races are decreasing; diverse land use management is inclining to be single and seasonal shifting grazing tradition is fading away. The change of diversity to singularity might cause some negative impacts on the local environment and ecosystem. In this paper, advices were also given on how to combine Tibetan traditional knowledge and management experiences into sustainable development of modern agriculture. In this thesis, genetic diversity of Musella lasiocarpa (Franch.) C. Y. Wu ex H. W. Li, a plant endemic to southwest China, was also discussed through the approach of SSR markers. The wild populations of M. lasiocarpa are very rare now due to the habitat fragment and long time human’s disturbance. By conducting broad field investigation, we have found 5 wild populations near the boarder of Yunnan and Sichuan province. Seventeen microsatellite markers were isolated from M. lasiocarpa by using FIASCO method. 8 primers were selected to do the further genetic population structure and genetic diversity analysis. The results showed that genetic diversity of M. lasiocarpa’s wild populations is higher than cultivated populations. The genetic diversity difference between wild and cultivated populations is related to the different reproduction systems. Adopting the way of asexuality reproduction, the genetic basis of cultivated populations become narrow that decrease the genetic diversity. AMOVA analysis showed that 37.19% genetic differentiation is among populations and 62.81% is within population. Genetic differentiation among different populations is related to the limited gene communication. POPGENE analysis indicated that there is very little gene flow among different populations (0.4916), which is the main reason of high genetic differentiation among M. lasiocarpa 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