| Lu Dangshen is the dry roots of Codonopsis pilosula(Franch.)Nannf.The top-grade medicinal materials of Codonopsis radix is produced in ancient Shangdang area in Shanxi Province(now Changzhi in Shanxi Province),hence the name.Codonopsis radix is a traditional tonic medicine,and is wildly used for both medicine and food.The market demand for Codonopsis radix is increasing.However,due to the environmental damage and the scarcity of wild resources caused by over excavation,the cultivated Codonopsis radix has become the main kind in the market.At present,there are some problems in the standardized production of Codonopsis radix,such as the confusion of germplasm,being easily affected by the environment,and extensive management of harvesting and processing,leading to the uneven quality of Codonopsis radix in the market.Therefore,efficient production of Codonopsis radix,improving its yield and quality,and studying its quality formation mechanism have become the key to its standardized production.In order to solve these problems,we have dug into the standardized production of Lu Dangshen,including the identification of three kinds of original products from adulterants by using DNA sequence;the metabonomics analysis of the differences among three kinds of Codonopsis and two ecotypes;the determination of the contents of 5 heavy metal residues and 19 other beneficial inorganic elements in Lu Dangshen and its rhizosphere soil by ICP-MS,and the analysis of their enrichment relationships;the research of mechanism of stress response by measuring the dynamic changes of physiological indexes,photosynthesis and chlorophyll fluorescence parameters in the growth process of Lu Dangshen under simulated temperature stress in an artificial climate chamber;the optimum harvest time,harvest time,drying method and rubbing process were optimized by multi index comprehensive evaluation method and the consistency of Codonopsis radix was evaluated by HPLC fingerprint.The main content of this subject include the following aspects:1.Study on germplasm resources of Codonopsis Radix.In this part,based on the ITS2 sequence,Codonopsis and other adulterants were identified by analyzing distance,mutation sites and phylogenetic tree.The chemical constituents of Codonopsis radix were analyzed by HPLC triple TOF MS/MS,and the metabolic profile was analyzed by multivariate statistical analysis method,with VIP>1,fold change>1.5 or<0.67,P<0.05.The results showed that 93,47 and 46 metabolites were obtained from C.pilosula and C.tangshen,C.pilosula and C.pilosula var.modesta,C.tangshen and C.pilosula var.modesta,and 19,18 and 12 metabolites were identified respectively MS/MS was used to analyze the chemical components of different ecotypes(wild type and cultivated type)of Codonopsis radix.The metabolic profile was analyzed by multivariate statistical analysis.The potential difference components were screened by VIP>1,fold change>1.5 or<0.67,P<0.05.49 difference metabolites were obtained and 14 difference metabolites were identified.2.Study on the effects of Soil and Temperature on quality formation of Lu Dangshen.In this part,microwave digestion combined with ICP-MS method was used to determine 24 inorganic elements in Lu Dangshen and its rhizosphere soil samples,and the content and enrichment coefficient were analyzed,according to which not only whether the content of heavy metals exceeded the standard was confirmed,but also the quality formation of Lu Dangshen was evaluated from the perspective of nutritional elements.The results showed that the content of heavy metals in 15 batches of Lu Dangshen was not exceeded,and the content of heavy metals in its rhizosphere soil was higher than the standard.The content of heavy metals also did not exceed the environmental soil standards.The medicinal materials of Lu Dangshen are rich in beneficial inorganic elements,including Mg and Fe.In addition,Lu Dangshen has a strong enrichment effect on Zn,which is closely related to the human immune system,and is so called "flower of life",and its main source of elements is rhizosphere soil.The artificial climate box was used to adjust the temperature to simulate the high temperature stress of plants,and the changes of physiological indexes,photosynthetic characteristics and chlorophyll fluorescence characteristics were detected to study the stress response mechanism of plants under high temperature stress.The results showed that high temperature stress led to a large amount of MDA accumulation,and the activities of CAT,SOD and POD first increased and then decreased Pn,Gs,Ci and Tr decreased significantly with the increase of stress time.Fo increased with the increase of high temperature stress time,Fv/Fm,ETR and QP decreased with the increase of temperature and stress time,which indicated that PS Ⅱ function of dangshen leaves decreased,photosynthetic electron transport capacity decreased and photosynthesis was inhibited in different degrees under high temperature stress.3.Study on the effect of harvesting and processing on the quality formation of Lu Dangshen.In this part,this study determined the contents of lobetyolin,atractylactone Ⅲ,syringol glycosides,and total polysaccharides of Lu Dangshen in different growth years and different harvest periods.Analyzed this contents change trend of each components in different harvesting periods.The contents of each component increases first and then decreases with the increase of growth time.It is best to choose two-year-old and harvest between October 20 and November 5 each year.In this study,fresh Lu Dangshen was used as raw material,and six drying methods were used to dry in the shade,sun-dried,infrared drying,microwave drying,vacuum drying,and hot air drying.The contents of lobetyolin,atractylactone Ⅲ,syringol glycosides,total polysaccharides and appearance traits of medicinal materials were used as evaluation indicators to assess the effects of different drying methods on the quality of Codonopsis Radix.Meanwhile,the best drying method of Codonopsis Radix was optimized by comprehensive evaluation of different drying methods combining with dry time and energy consumption.It shows that the optimal drying method for Codonopsis Radix was infrared or hot air drying at 60℃.This study adopts orthogonal test design.We taked the drying temperature,the water content before kneading,the number of kneading and the time of each kneading as the inspection factors,and used lobetyolin,Atractylaceton Ⅲ,syringol glycosides,and total polysaccharides of Lu Dangshen as indicators,and combined with the comprehensive scoring method to screen and optimize the rubbing and drying process of Lu Dangshen.And we maked a preliminary jjudgment.The drying temperature and the number of kneading were relatively important in the kneading and drying process.The best kneading and drying process was preliminarily selected as the drying temperature of 50℃ the water content of 30%before the first kneading,the kneading twice,and each time 3 minutes each time.4.Study of fingerprints of Lu Dangshen.In order to evaluate the quality of Lu Dangshen more comprehensively and objectively.This study used HPLC to study the fingerprints of 15 batches of cultivated Lu Dangshen,established its fingerprint spectrum conditions.Its calibrated 14 common peaks on the Similarity evaluation system for chromatographic fingerprint of Traditional Chinese Medicine.The comparison of the reference substance was used to confirm the lobetyolin peak and the atractylenolide Ⅲ peak,and we were calculated the relative retention time,relative peak area and similarity of each shared peak.The similarity of each sample is greater than 0.90.This shows that the consistency between batches of samples is better. |
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