Comparison And Analysis Of Genetic Diversity And Quality Of Rhodiola

There are 32 species and two varieties of Rhodiola in Tibet Autonomous Region, accounting for about one-third of the world’s.Because of its high medicinal value,and its being an edible plant,Tibet Rhodiola is mainly used as an raw material in clinical treat, health food, cosmetic and chemical aspects in large quantities;it is also known as "plateau ginseng" for its high degree of popular concern.At the same time,Rhodiola is also used as a gift for friends and families to share as a plateau specialty;more importantly,Rhodiola can be used for defense training, production, construction, tourism and other aspects in order to ease the high altitude discomfort.However,due to natural climate such as low temperature,low-oxygen content,strong ultraviolet radiation,and large temperature difference between day and night,the growth of Rhodiola is quite slow;thus,in 2009 the Rhodiola was listed as an endangered.Tibetan medicinal species in Tibet Autonomous Region. How to tackle the problems of its high demand and supply shortage are main subjects in this research.Research on the genetic diversity of Rhodiola:Researchers of this project have visited, investigated resources and collected samples in Lhasa, Shannan, Changdu, Nyingchi and Shigatse for study on its genetic diversity. The result shows that the growing region of Rhodiola is changing year by year, presenting a dynamic distribution; the natural environment has a decisive constraint on the growth of Rhodiola. This study on the one hand provides a macroscopic guidance to determine the distribution of Rhodiola in future, and on the other hand calls for people to raise the awareness to protect the natural environment.The growth of new branches, roots and the blossom of the isolated shoot taken from R.fastigiata in Nyingchi, R.serrata H.Ohba in shannan, R.himalensis in Shigatse are observed and studied by researcher of this project in Kunming under the condition of room temperature.Researchers took about 10cm tip of Rhodiola from the collected branch samples, put it in a beaker and added 100ml water to cultivate. We took different individual genome DNA in new shoots, used generic primers ITS1 for PCR amplification, and found a high genetic diversity in four kinds of rhodiola rosea; simultaneously.we conducted an ITS sequencing, molecular identification and analysis of three complete segments of Rhodiola, and the result suggests that the distribution of Rhodiola in that area has a high genetic diversity. In addition, the genetic diversity of Rhodiola found in this paper provides not only a technical basis and effective primer to further analyze its genetic diversity, but also a foundation for future design and distinguish between Rhodiola and other kinds of specific PCR primers.On the aspect of quantitative evaluation:in the qualitative and quantitative analysis of chemical composition, researchers use HS-GC-MS on 12 samples to analysise their volatile components,30 chemical compounds were identified from them. It was found that the similarity rate of ion flow diagram is above 0.9 of the same origin Rhodiola in different region. R.fastigiata is very similar to rhodiola crenulata, and the similarity rate between R.serrata H.Ohba, R.himalensis, R bupleuroides and Rhodiola crenulata decreases in turn.To determine the content of Rhodiola salidroside and gallic acid in Rhodiola samples by HPLC found a big difference of salidroside in Rhodiola in different regions. There are no salidroside in one samples, but there is Gallic acid in each sample, and there is no regularity of their content.In LC-MS/MS method to analysize the main chemical components in Rhodiola crenulata, the result shows a difference in chemical composition of Rhodiola crenulata in different regions.The author of this paper analyses and studies on six active components in Rhodiola crenulata from different Tibetan habitats to establish a quantitative analysis method, and provides a basis for its further use. Methods:take samples with 30% ethanol; Diamonsil C18 chromatographic column (250×4.6mm),conduct a gradient elution with acetonitrile(A)-0.3% phosphoric acid solution (B),detect wavelength 275nm and the column temperature is 25℃. Results:gallic acid, salidroside, tyrosol, catechin, gallic acid ethyl ester, coumaric acid are in sound linear relationship, they range from 0.0382 μg.ml-1～0.382 μg.ml-1 (r= 0.9998),0.301 μg.ml-1～3.01 μg.ml-1 (r= 0.9999), 0.0198μg.ml-1～0.198μg.ml-1(r= 0.9996), 0.0171 μg.ml-1～0.171 μg.ml-1 (r=0.9996),0.0045μg.ml-1～0.045μg.ml-1(r= 0.9998),and 0.00638μg.ml-1～0.0638 μg.ml-1(r=0.9994) respectively and all the rates of recovery are in line with the requirement of content determination. Conclusion:the method is simple, rapid and accurate, and it can be used to detect the content of six active ingredients in Rhodiola.The author also determines the content of polysaccharide in Rhodiola from different origins in Tibetan Plateau by UV spectrophotometric.Methodising sulfuric acid—nthranone to detect 617nm wavelength.Result:the linear range is from 1.0Oμg.ml-1～200.0μg.ml-1(R= 0.9999).Conclusion:this method is convenient,rapid, accurate and reliable,and can be used as a method for the determination of polysaccharide in Rhodiola.As toxicity in drug effect is an important index to evaluate the quality of drugs, therefore, the author conducts a comparative study on the pharmacodynamics and toxicity of rhodiola rosea,on the form of thrombosis and hemorrheology,on hypoxia tolerance test,and on the influence of older mice’s immunity and anti-oxidation function.The results show that on aspects of improving immune function and hemorrheology, Rhodiola crenulata can be replaced by R.fastigiata; on aspect of enhancing body’s hypoxia tolerance, R.serrata H.Ohba can be used as a substitute of Rhodiola crenulata. The research on the acute toxicity of three kinds of rhodiola shows there is no obvious toxicity and they are in the safety scope.