Correlation Between Main Effective Components In Two Species Of Dasyphora And Ecological Factors

Human use medicinal plants to treat and prevent disease for thousands of years history.Today, about25%of prescription drugs generate from the medicinal plants.Most of the active ingredients in medicinal plants are plant secondary metabolites. Secondary metabolites regulated commonly by the genetic characteristics (internal) and plant growth environment (external) are the non-essential small molecule compounds in cellular activities or the proper functioning of the plant growing development. Its generation and distribution usually has obvious geographical, species, organ, tissue and reproductive period specificity, while environmental changes can affect the types and quantities of secondary metabolites directly, or synthesize specific secondary metabolites in the particular environment and time conditions only. Therefore, any kind of natural ecological factors in the environment are likely to affect the synthesis and accumulation of secondary metabolites in plants. In contrast with the primary metabolism (metabolites), secondary metabolism (metabolites) has stronger correlation and correspondence with the environment.For this reason, to determine the dominant environmental factors for the synthesis and accumulation of the plant secondary metabolites is the key to guarantee the quality of wild medicinal plants and improve the efficacy of medicine. However, in the actual production, quality and yield of medicinal plant are the two conflicted aspects, since to improve the content of secondary metabolites is bound to lose part of the production. Consequently,This thesis is focus on studying the role and role size of the main ecological factors on quality of Chinese herbal medicine to reveal the dominant factors. Thus, artificially regulating and balancing the contradiction between quality and yield.Potentilla fruticosa, and Potentilla glabra (both in Rosaceae), are one of the earliest applications in traditional Chinese medicine, with heat clearing, heart purifying and brain beneficial, digestion promoting, spleen invigorating and dampness eliminating, menstruation regulating functions, etc. They are typical deciduous shrub in the alpine region in China, located in Xinjiang, Qinghai, Shaanxi, Gansu, Ningxia, Inner Mongolia, Sichuan, Shanxi, Heilongjiang, Jilin Province, etc. The thesis selected eight representational plots, Huzhu Northern Mountain in Qinghai Province Taibai Mountain, Zibai Mountain, Huoditang, and Niubeiliang in Shaanxi Province, Emei Mountain in Sichuan Province, Yunding Mountain in Shanxi Province, Helan Mountain in Inner Mongolia Autonomous Region as the research object, studying the major active ingredients, in vitro antioxidant activity, climatic conditions of the sampled plots, and soil conditions of Potentilla fruticosa and Potentilla glabra, aiming to ascertain the correlation of the active component content and antioxidant activity with all the ecological factors of them, screening the dominant factors, and providing a theoretical basis for the protection, utilization and sustainable management of Potentilla fruticosa and Potentilla glabra.1. Optimization of the main secondary metabolites extraction process of Potentilla fruticosa and Potentilla glabra leavesOn the basis of the single-factor test, according to principle of the Box-Behnken central composite design, using the response surface design to optimize the extraction process of the tannin and total flavonoid of Potentilla fruticosa and Potentilla glabra leaves. According to the analysis of Design-Expert software, the optimized extraction process for Potentilla fruticosa were as follows:ethanol concentration of70%, extraction temperature of80℃, extraction time of2h, extraction times of3, the liquid ratio of20:1. The optimized extraction process for Potentilla glabra were as follows:ethanol concentration of90%, extraction temperature of80℃, extraction time of2h, extraction times of3, the liquid ratio of15:1.2. Differentiation of the effective components and antioxidant activity of Potentilla fruticosa in the two representative plotsThis experiment selects two typical plots—Potentilla fruticosa from Huzhu Northern Mountain in Qinghai Province and Taibai Mountain in Shaanxi Province, measuring the effective components content and antioxidant activity of the ethanol extracts of different parts of them. Tannin and rutin content of Potentilla fruticosa leaves from Huzhu Northern Mountain in Qinghai Province are the highest, reaching8.04%and0.46%, respectively. Flavonoid content of Potentilla fruticosa flowers from Taibai Mountain in Shaanxi Province is the highest, reaching8.46%.It is proven that leaves has the strongest antioxidant activity. Potentilla fruticosa leaves from Huzhu Northern Mountain in Qinghai Province have the strongest antioxidant activity among them, with the DPPHIC50value at7.32μg/ml. While compared with the antioxidant capacity of rutin standard product(DPPHIC50=5.79μg/ml), it can be seen that there is no significant differences between them(P<0.05). The results of this study show that significant differences (P<0.05) between the effective components content and antioxidant activity of different parts of Potentilla fruticosa in different plots. the effiective components content and antioxidant capacity of Potentilla fruticosa leaves from Huzhu Northern Mountain in Qinghai Province is the highest among them. The reason for this difference may be:(Ⅰ) Plant’s own metabolic factors.(Ⅱ) Ecological factors such as soil and climate.(Ⅲ) Genetic mutations. This result is able to provide the theoretical basis for using Potentilla fruticosa leaves as natural antioxidants and traditional Chinese medicine.3. Correlation of the active components and antioxidant activity of Potentilla fruticosa leaves with ecological factorsIn this study, the impact of ecological factors on tannin, flavonoid, rutin content and antioxidant activity of the Potentilla fruticosa leaves are studied using correlation analysis, multiple linear stepwise regression analysis, and path analysis. Rapidly-available phosphorus, total potassium, and annual lowest temperature are the dominant factors affecting the tannin content, among which rapidly-available phosphorus is a direct effective factor, presenting a significant negative correlation with the tannin content; soil pH and annual average temperature are the dominant factors affecting the total flavonoid content, among which pH is a direct effective factor, presenting a significant positive correlation with the total flavonoid content; total phosphorus, annual sunshine hours and frost-free period are the dominant factors affecting the rutin content of, among which annual sunshine hours is a direct effective factor, presenting a significant negative correlation with the rutin content; rapidly-available phosphorus, organic matter and annual average temperature are the dominant factors affecting the antioxidant activity, among which annual average temperature is a direct effective factor, presenting a significant positive correlation with antioxidant activity. The results show that the ecological factors are principle elements affecting active components content and antioxidant activity of Potentilla fruticosa leaves. Though, the dominant factors affecting various active components and antioxidant activity capacity are different, they are mainly focused in the rapidly-available phosphorus, pH, annual sunshine hours, and annual average temperature.4. Correlation of the active components and antioxidant activity of Potentilla glabra leaves with ecological factorsThe correlation of ecological factors on the tannin, flavonoid content and in vitro antioxidant activity of Potentilla glabra leaves in six different plots are analyzed using the gray relational analysis. The results show that there is significant difference in the quality of Potentilla glabra in different plots, with the tannin content between4.95%～7.74%; the flavonoid content between1.07%～3.56%, among which the tannin and flavonoid content of Potentilla glabra leaves from Huzhu Northern Mountain in Qinghai are the highest, reaching7.74%and3.56%, respectively; Potentilla glabra leaves from Helan Mountain have the strongest antioxidant capacity, with its DPPHIC50value at6.9g·ml-1; in the gray correlation analysis, annual lowest temperature, ammonium nitrogen, frost-free periods, organic matter, rapidly-available potassium, and annual highest temperature are the dominant factors affecting the tannin content; annual sunshine hours, annual lowest temperature, pH, effective accumulated temperature, organic matter, and total nitrogen are the dominant factors affecting the flavonoid content; annual sunshine hours, annual average rainfall, organic matter, pH, annual lowest temperature, and annual highest temperature are the dominant factors affecting the antioxidant activity.