| In recent years, with the increasing of the market demand and changes in the environment, Tibetan medicine resources was sevciously damaged. Finding out the distribution of resources and protect the wild resources is particularly urgent. On the other hand, the phenomenon of synonymies and homonymies in Tibetan medicine, resulting in varieties identification difficult, seriously impact the quality control and evaluation. Therefore this paper aims at the complex multi original Tibetan medicine "Dida" as the research object, focused on its species identification by DNA barcode; Besides, then carry out Hippophae rhamnoides L.subsp. sinensis Rousi’resources investigation, normalized planting, ecological suitability zoning, accumulation of chemical components and ecological factors,to provide scientific basis for variety identification, the protection of wild resources and the promotion of artificial cultivation in the suitable habitat.Objectives:1. To identify Tibetan medicine "Dida" by DNA barcoding; To select the suitable habitat Hippophae rhamnoides L. subsp. sinensis Rousi. and to investigate the dominant ecological factors whith affect the accumulation of chemical components.Methods:1. ITS2 sequence was Used to identify different of "Dida", to provide the basis of varieties identification from the molecular level.2. Based on the Maxent model, the suitable area is studied;3. TLC was used to identify, the contents of flavonoids were determined by HPLC:4. Pearson correlation analysis, stepwise regression analysis and partial least squares (PLS) were used to analyze the correlation between the chemical components and their ecological factors.Results:1.ITS2 sequence can be used for "Dida" identification. ITS2 sequence length were 231 bp, GC content was an average of 61.40%, Phylogenetic analysis demonstrated that every species clustered into their own clade, except Swertia bifolia and Swertia wolfangiana clade2.Widely distributed in the provinces of China, wide distribution area, large number of resources. Like the sun, for the water has certain requirements. Often with shrubs, grass shrub vines formed structure.Established 3 standardized cultivation.demonstration base, a total of 6 acres.3.China’s regional distribution of the suitable index for 0-0.708. suitable area 59.05 ten thousand km~2, accounting for 6.13% of the total area. More suitable area 55.25 ten thousand km", accounting for 5.73% of the total area.With quercetin, kaempferol and isorhamnetin as controls, Determination of the content of 17 batches by HPLC:quercetin 0.1929%-0.5236%, isorhamnetin 0.2242%-0.6236%, kaempferol 0.0772%-0.1775%. Results show that the content difference among different producing areas.4. Determine the dominant ecological factor and it’s optimal range that effect the accumulation of quercetin,kaempferol and isorhamnetin.Effect of quercetin content accumulation:altitude is 3009-3320m; the average monthly precipitation in January and August are 4.0-6.0mm and 118-152mm respectively.Effect of kaempferol accumulation:altitude is 2999-3315m;the aerage monthly precipitation in the coldest quarter and December are -9.0-0.1℃. Effect of isorhamnetin accumulation:the average monthly precipitation in August,January and the coldest quarter are118-122mm,4.0-5.0mm and 13.0-22.0mm;altitude is 3158-3568m.Conclusions:1. ITS2 sequence as the Gentianaceae Tibetan medicine Dida’s DNA barcode with good accuracy.2. Hippophae rhamnoides L. subsp. sinensis Rousi is widely distributed in china.with a wide area and a large number of resources, which is suitable for sustainable development and utilization. |
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