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To solve the serious problem of stem and leaf shading in the middle and late stage of traditional flat planting of Codonopsis pilosula, this study analyzed the effects of different stereoscopic traction heights on the photosynthetic characteristics and growth of C. pilosula and explored the optimal traction height to improve the yield and quality of C. pilosula. The experiment designed three stereo-scopic traction heights [H1(60 cm), H2(90 cm), and H3(120 cm)] with natural growth without traction as the control(CK). The results showed that the increase in stereoscopic traction heights broadened the growth space of stems and leaves of C. pilosula, enhanced the ventilation effect, significantly increased the average daily net photosynthetic rate of C. pilosula, promoted the absorption of intercellular CO_2, decreased the transpiration rate, and reduced the evaporation of water. Moreover, it effectively avoided the problem of weakened photosynthesis, maintained the carbon balance of individual plants, and promoted the growth and development of the C. pilosula roots. In terms of the seed yield of C. pilosula, it was ranked as H2>H1>H3>CK. To be specific, H1 increased by 213.41% compared with CK, H2 increased by 282.43% compared with CK, and H3 increased by 133.95% compared with CK. The yield and quality of C. pilosula were the highest in the H3 treatment group, with the fresh yield of 6 858.33 kg·hm~(-2), 50.59% higher than CK, dry yield of 2 398.33 kg·hm~(-2), 76.54% higher than CK, and lobetyolin content of 0.56 mg·g~(-1), 45.22% higher than CK. Therefore, the stereoscopic traction height has a great influence on the photosynthetic characteristics, yield, and quality of C. pilosula. Particularly, the yield and quality of C. pilosula can be optimized and improved in the traction height treatment of H3(120 cm). This planting method is worth popularizing and applying in the cultivated management of C. pilosula.
Subject(s)
Codonopsis , Traction , Photosynthesis , Plant Leaves , Plant RootsABSTRACT
ObjectiveThe transcriptome characteristics of different tissues of Codonopsis pilosula were analyzed to illustrate the genetic basis of the accumulation of active ingredients in the root of C. pilosula， and to provide theoretical basis for its high-quality production and cultivation. MethodDifferent tissues of C. pilosula at flowering stage were selected as experimental materials， and the contents of tangshenoside Ⅰ， lobetyolin and atractylenolide Ⅲ were detected by high performance liquid chromatography（HPLC）. RNA-Seq was used to perform transcriptome sequencing of different tissues， and the differentially expressed genes were screened and analyzed by Gene Ontology（GO） and Kyoto Gene and Encyclopedia of Genes and Genomes（KEGG） enrichment analysis， in order to explore the characteristics of active compound distribution and the transcriptional profiles. ResultThe contents of polysaccharides and tangshenoside Ⅰ in the root of C. pilosula were significantly higher than those in other tissues. The transcriptional profiles of the root were significantly different from those of stem， leaf and flower. Cluster analysis， GO and KEGG enrichment analysis of differential gene expression showed that the differential expression genes were mainly enriched in flavonoid and phenylpropanoid biosynthesis， sucrose-starch metabolism， plant hormone signal transduction， plant-pathogen interaction， mitogen-activated protein kinase（MAPK） cascade signal transduction， Adenosine triphosphate（ATP）-binding cassette（ABC） transporter and other pathways. The expression of genes related to biosynthesis of phenylpropanoid compounds were significantly up-regulated in the roots and flowers， and ABC transporter proteins were mostly highly expressed in the flowers. The expression of key enzyme genes for polysaccharide synthesis， such as sucrose：sucrose 1-fructosyltransferase（1-SST） and fructan 1-exohydrolase（1-Feh）， were significantly up-regulated in the roots， and a large number of stress-responsive genes closely related to the accumulation of secondary metabolites were significantly up-regulated in the roots. ConclusionThe active compound content and transcriptional profiles in C. pilosula roots were significantly different from those in stem， leaf， flower and other tissues， showing tissue specificity. Meanwhile， the genes related to stress response and biosynthesis of active compound， such as fructan and phenylpropanoid compounds， were up-regulated in roots of C. pilosula.
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Lu Dangshen, a traditional authentic medicinal material of Codonopsis Radix is mainly produced in Shangdang (Changzhi) area of Shanxi Province. Baitiao Dangshen is mainly produced in Gansu Province. Codonopsis Radix contains many kinds of components such as phenylpropanoids, polyalkynes, alkaloids, terpenes, fatty acids, flavonoids, and so on. At present, the effect of producing areas on its chemical compositions has not been systematically studied. This study analyzed the differences of metabolites among Codonopsis pilosula from different producing areas by UPLC-HRMS. PCA, OPLS-DA coupled with Thermo mzcloud online and local databases were used to compare the overall differences of metabolites among Codonopsis pilosula from different producing areas, and the chemical constituents were identified to further screen and find out the different metabolites and analyze the metabolic pathways by information retrieval in HMDB, PubChem, Chemspider and KEGG databases. The results showed that 72 differential metabolites were identified in this study. There were 15 kinds of up-regulated and 57 kinds of down-regulated metabolites of Lu Dangshen compared with Baitiao Dangshen. The top 30 metabolic pathways were analyzed by KEGG enrichment, and the most important metabolic pathways were phenylpropanoid biosynthesis, which was demonstrated that phenylpropanoid biosynthesis pathway and related intermediate metabolites could be used as the characteristics of distinguishing Lu Dangshen from different habitats of Codonopsis pilosula. The present study provided a basis for analyzing the influence of producing areas on the chemical components of Codonopsis pilosula and reasonably evaluating the quality of Codonopsis Radix, and also provided a new idea for expounding the authenticity of Lu Dangshen.
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OBJECTIVE To study the quality of Codonopsis pilosula with different commodity specification grades， and to provide the data support for market transactions， scientific research and clinical use. METHODS According to the classification standard of commodity specification grades of C. pilosula， 17 batches of C. pilosula from different producing areas， origins and commodity specification grades were collected. The contents of tangshenoside Ⅰ， lobetyolin and atractylenolide Ⅲ were determined by HPLC. The contents of alcohol-soluble extracts were determined by hot dipping method stated in general rule 2201 of Chinese Pharmacopeia （part Ⅳ）. The contents of polysaccharide were determined by phenol-sulfuric acid method （calculated by D-glucose anhydrous）. RESULTS For cultivar of C. pilosula， four specifications and three commodity grades of C. pilosula all contained tangshenoside Ⅰ and lobetyolin； Radix C. pilosula from Shanxi of China and C. pilosula from Wenxian County of China， also contained atractylenolide Ⅲ. In terms of the contents of tangshenoside Ⅰ， lobetyolin and atractylenolide Ⅲ， the content of second class was equivalent to that of first class， even better than the first class， while the content of third class was lower than that of first class and second class； the content of tangshenoside Ⅰ was the highest among the two types of wild C. pilosula. The contents of alcohol-soluble extracts and polysaccharides in first class cultivated C. pilosula were higher than those of second class， and the second class was higher than the third class； wild C. pilosula had low content of alcohol-soluble extracts and polysaccharides. CONCLUSIONS The internal quality of C. pilosula is basically consistent with the classification standard of different commodity specification grades； the content of each indicator in first-class and second-class medicinal herb is high， making them high-quality medicinal herbs.
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We performed an extensively targeting metabolomic detecting using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) to compare the secondary metabolites in Dang shen [Codonopsis pilosula (Franch.) Nannf.] from Shanxi and Gansu provinces. The findings showed that 161 secondary metabolites in 6 groups (phenolic acids, flavonoids, lignans and coumarins, alkaloids, terpenoids, others) were found from Dang shen in Changzhi city of Shanxi province and Dingxi city of Gansu province. There were 98 secondary metabolites which is differed significantly. In comparison to Dingxi city, 33 different secondary metabolites of Dang shen in Changzhi city had a greater relative content, whereas relative content of 65 different metabolites in Dingxi city was higher. Metabolic pathway enrichment analysis revealed that phenolic acids and flavonoids were significantly different in the secondary metabolites of Dang shen from different producing places. This may be one of the reasons for the difference in the quality of Dang shen in Shanxi and Gansu provinces. This work compared and analyzed the secondary metabolites of Dang shen from Dingxi city in Gansu province and Changzhi city in Shanxi province for the first time, which lays the foundation for further study on the quality of Dang shen.
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ObjectiveTo explore the correlation between the content of 4 functional components in Codonopsis pilosula roots from different areas and soil factors， and thereby to lay a theoretical basis for soil ecological regulation and improvement of quality of C. pilosula roots. MethodThe content of lobetyolin， atractylenolide Ⅲ， alcohol extract， and polysaccharides， as well as soil fertility and 16 soil factors in 24 batches of samples from different producing areas were determined. Principal component analysis （PCA） and Pearson's correlation analysis were used to explore the key soil factors leading to the variation of chemical component content in C. pilosula roots. ResultThe content of lobetyolin and atractylenolide Ⅲ in samples from Longxi was the highest， and the content of polysaccharides peaked in samples from Huguan. The content of lobetyolin was in positive correlation with soil total nitrogen， total phosphorus， total potassium， alkali-hydrolyzable nitrogen， and available potassium （P<0.01）， as well as soil organic matter， pH， available manganese， and available zinc （P<0.05）. There was a positive correlation between pH and atractylenolide Ⅲ content （P<0.05）. Soil total potassium was in positive correlation with alcohol extract and polysaccharide content （P<0.01）. Soil available zinc was positively correlated with alcohol extract and the polysaccharide content （P<0.05）. Sample sites with higher PCA scores were Pingshun， Huguan， and Longxi， which were significantly positively correlated with the content of polysaccharides in C. pilosula roots in different habitats. ConclusionThe content of functional components in C. pilosula roots can be improved by raising soil organic matter content and applying specific fertilizers.
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Lu Dangshen is the geoherb in Shanxi Province. The content of Codonopsis pilosula polysaccharides (CPP) in Lu Dangshen is more than that in other Codonopsis Radix from other regions. Glycosyltransferase is the key enzyme for the synthesis of bioactive components, such as CPP and tangshenoside I. Based on the transcriptome data of C. pilosula [Codonopsis pilosula (Franch.) Nannf.] from different producing areas, this study carried out functional annotation of GO and KEGG, conservative domain analysis, phylogenetic tree analysis and expression pattern analysis of glycosyltransferase genes in C. pilosula to provides a theoretical basis for exploring the mechanism of genuineness formation in Lu Dangshen. In this study, 98 glycosyltransferase genes were screened and identified, which belonged to GT family 1, GT family 2, GT family 90 and other families. By GO functional annotation, it was found that most of the glycosyltransferase genes had catalytic activity. Analysis of KEGG functional annotation showed that C. pilosula glycosyltransferase was mainly involved in glycan organism and terpenoid and polyketone metabolism. Among them, conserved domain of 42 glycosyltransferase genes in GT family 1 was [X]-W-[2X]-Q-[3X]-[LH]-[5X]-[FLTHCGWNS]-[2X]-E-[4X]-[GVP]-[4X]-P-[4X]-Q-[2X]-[NAK]. Phylogenetic tree analysis based on the glycosyltransferase sequence in Arabidopsis thaliana showed that C. pilosula glycosyltransferases were mainly located in Arabidopsis thaliana UGT73, 72 and 85 branches. Gene expression pattern analysis showed that expression of CpUGT73AH2 was higher in Lu Dangshen than that in Baitiaodang and could respond to drought and low temperature stress. In conclusion, a glycosyltransferase gene CpUGT73AH2, which is involved in the metabolism of terpenoids and polyketides and can respond to environmental stress, was screened from the C. pilosula glycosyltransferase family 1, which was used to further study the role of C. pilosula glycosyltransferase in Lu Dangshen. It laid a theoretical foundation for further study on the role of C. pilosula glycosyltransferase in the formation of Lu Dangshen.
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OBJECTIVE： To study the tonifying spl een effect of Codonopsis pilosula polysaccharides （CPP） on spleen-deficiency model mice. METHODS ：Totally 60 ICR male mice were randomized into blank group ，model group ，CPP high-dose，medium-dose and low-dose groups [ 1.6，0.8，0.4 g/（kg·d）]，Sijunzi tang （SJZT）group [ 30 g/（kg·d），by crude drug] ， with 10 mice in each group. Except for blank group ，other groups were given Sennae Folium solution intragastrically （0.4 g/d）to establish spleen-deficiency model. After modeling ，administration groups were given relevant drug intragastrically ，and blank group and model group were given 20 mL/kg normal saline intragastrically ，once a day ，for consecutive 6 weeks. After last administration，body weight of mice in each group was measured and their general behavioral characteristics were observed. The red blood cell count in whole blood were recorded ，and the contents of D-xylose，gastrin（GAS），motilin（MTL），somatostatin （SS），vasoactive intestinal peptide （VIP），amylase（AMS），immunoglobulin（IgG and IgM ）and lipopolysaccharide （LPS）in serum were determined ；mRNA and protein expressions of Claudin and Occludin in colon tissues of mice were also detected. RESULTS：Compared with blank group ，body weight of mice in model group was significantly decreased （P＜0.05），and the spleen deficiency symptoms such as sparse back hair and no luster appeared ；the red blood cell count in whole blood ，serum contents of D-xylose，SS，VIP，AMS，IgG and IgM ，mRNA and protein expressions of Claudin and Occludin were significantly decreased（P＜0.05 or P＜0.01）；serum contents of GAS ，MTL and LPS were significantly increased （P＜0.01）. Compared with model group ，the body weight of mice were increased significantly in CPP high-dose group （P＜0.05），and spleen-deficiency symptom was improved significantly. Except for the serum contents of D-xylose and IgM ，the protein expressions of Claudin and Occludin in colon tissue had no statistical significance in CPP low-dose group （P＞0.05），above indexes of other groups w ere improved significantly （P＜0.05 or P＜0.01）. CONCLUSION ：CPP can i mprove spleen-defic iency model mice induced by Sennae Folium，the mechanism of which may be associated with regulating gastrointestinal hormone secretion ， enhancing immunity and protecting intestinal mucosal barrier.
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Objective:To explore the high-efficiency and high-quality seedling raising method of <italic>Codonopsis pilosula</italic>. Method:In the main production area of <italic>C. pilosula</italic> in the Tanchang county，Gansu province，after the soil was fumigated with dazomet （containing 98% methylisothiocyanate）， four varieties of <italic>C. pilosula</italic> seedlings were raised. The dynamic change in growth and differences in quality and yield of <italic>C. pilosula</italic> seedlings after emergence were regularly determined. Result:① The soil enzyme activity was first inhibited and then restored by soil fumigation，which increased the root length of <italic>C. pilosula</italic> seedlings by 9.8%. Besides， the field growth indexes such as plant height，plant width，stem length，stem diameter，number of branches，number of nodes，number of leaves， and fitted leaf area increased in varying degrees，and the plant height showed an "S"-shaped growth trend. "Gandang No.1" and "Gandang No.2" grew better than "Weidang No.1" and "Tanchang control". ② Fumigation reduced the incidence rate of <italic>C. pilosula</italic> root in the field by 4.9%，and the incidence rates of "Gandang No.1" and "Gandang No.2" were significantly lower than those of "Weidang No.1" and "Tanchang control". ③ Fumigation increased the total number of <italic>C. pilosula </italic>seedlings by 6.15×10<sup>5</sup> plants·hm<sup>-2</sup>，of which the number of primary seedlings increased by 45.3% and that of secondary seedlings increased by 42.2%. ④ Fumigation increased the seedling yield of <italic>C. pilosula</italic> by 42.4%. It showed the most significant effect on the yield of "Gandang No.2"，which increased by 61.8%， and the weakest effect on the yield of "Gandang No.1"，which increased by 15.4%. ⑤ Comprehensive analysis showed that the quality and yield of <italic>C. pilosula</italic> seedlings in the fumigation area were better than those in the non-fumigation area. Conclusion:The results showed that soil fumigation showed a promoting effect on the seedling yield of <italic>C. pilosula</italic> in spite of different effects achieved in terms of different varieties.
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Objective:The biological mechanism of <italic>Codonopsis pilosula</italic> adaptation to drought was explored by determining the root metabolome of <italic>C. pilosula</italic> during harvesting. Method:Non-targeted metabonomics LC-MS was used to screen differential metabolites by multivariate statistical analysis，univariate statistical analysis and metabolic pathway enrichment analysis. Result:①There were 274 metabolites in LD vs CK group，142 of which were up-regulated and 132 of which were down-regulated. There were 284 metabolites with significant difference in MD vs CK group，of which 157 were up-regulated and 127 were down-regulated. There were 317 metabolites with significant difference in SD vs CK group，of which 133 were up-regulated and 184 were down-regulated. ②Differential metabolites were annotated into kyoto Encyclopedia of Gene and Genomes （KEGG） database and 82 differential metabolic pathways were obtained，among which sphingolipids metabolism was significantly enriched （<italic>P</italic><0.01）.Metabolism of arginine and proline，tryptophan，alanine，galactose，nicotinic acid and nicotinamide，cysteine and methionine，arachidonic acid，linolenic acid and glycerides were significantly enriched in different metabolite pathways （<italic>P</italic><0.05）. ③The metabolites of the three comparison groups before and after enrichment were classified and analyzed. It was found that they were mainly concentrated in fatty acyls group，carboxylic acid and derivatives，and organ oxygen compounds，followed by sphingolipids，indoles and derivatives，organonitrogen compounds，glycerophospholipids，pyridines and derivatives，peptidomimetics，glycerolipids and so on.In the drought stress of <italic>C. codonopsis</italic>，carbohydrate related metabolites were mainly up-regulated，lipid related metabolites were mainly down-regulated，and all other metabolites were up-regulated. Conclusion:The changes of metabolites in the roots of <italic>C. pilosula</italic> under drought stress were elucidated. carbohydrate and lipid-related metabolites were the main products of <italic>C. pilosula</italic> under drought stress，and these metabolites may be the main reason to improve the ability of <italic>C. pilosula</italic> to resist drought，which laid a foundation for further study on the mechanism of <italic>C. pilosula</italic> to resist drought.
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Objective:To establish the grade evaluation method for Codonopsis Radix slices based relative quality constants， in order to provide scientific theoretical basis for grading of Codonopsis Radix slices. Method:Through literature and market research，the main production areas of Codonopsis Radix slices were determined，and 67 batches of Ludangshen slices（52 batches） and Baitiaodang slices （15 batches） were collected. The appearance traits （average quality and average thickness of Codonopsis Radix slices） were observed and measured. According to Chinese Pharmacopoeia （2015 edition）， the extract and the content of Codonopsis pilosula polysaccharide was determined by phenol-sulphoacid method. Then the relative quality constant was calculated，and the results of grade evaluation were evaluated through systematic cluster analysis and correlation analysis. Result:Relative quality constants of 67 batches of Codonopsis Radix slices were between 0.32-2.97. If these samples were divided into three grades：the first-grade relative quality constants were greater than or equal to 2.08，the second grade was greater than or equal to 0.89 but less than 2.08，while the third grade was less than 0.89. The results of systematic cluster analysis showed that 67 batches of Codonopsis Radix slices were clustered into 3 categories，and the results were basically consistent with the classification. The correlation analysis showed a positive correlation between the content of the extracts and the polysaccharide content （P<0.05）. Conclusion:This method links the extrinsic characteristics to the intrinsic quality，and objectively grade Codonopsis Radix slices， so as to provide a basis for its grade standards.
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Based on 1H NMR metabonomics combined with multivariate statistical analysis, to study the changes of serum endogenous metabolites from immunosuppressive rats induced by cyclophosphamide (CTX), and to screen the best processed products of Codonopsis pilosula, rice-fried Codonopsis pilosula and honey-fried Codonopsis pilosula in improve immunity, and to explore its mechanisms. Methods CTX was administered intraperitoneal (i. p.) to induce the immunosuppressive model. Serum was collected by aorta abdominalis on the last day for detecting immunocyte, immunokine and 1H NMR analysis. Results Codonopsis pilosula and its processed products could increase the numbers of white blood cells and B lymphocytes, as well as the ratio of CD4+ and CD8 + T cells. The regulation effects of honey-fried Codonopsis pilosula and Codonopsis pilosula on IL-2, IgG and sigA were better than those of rice-fried Codonopsis pilosula. Honeyfried Codonopsis pilosula could significantly improve the levels of leucine, isoleucine, alanine, glutamate, glutamine, glucose and valine etc. Conclusions Honeyfried Codonopsis pilosula could play the best role in immunosuppressive rats through regulating valine, leucine and isoleucine metabolism, glutamate and glutamine metabolism, alanine, aspartate and glutamate metabolism.
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The medication rules of high frequency herb-pairs containing Codonopsis pilosula (Dangshen) were analyzed with data mining tools, and the molecular mechanisms of these herb-pairs on the gastrointestinal diseases were predicted with the network pharmacology. The R language association rules were used to mine the high frequency herb-pairs from TCM formulae containing Dangshen, and these herb-pairs would be screened out, which satisfied the following requirements with support ≥ 0.3 and confidence ≥ 0.9 at the same time. Using the Integrated Pharmacology Platform of Traditional Chinese Medicine (TCMIP) to predict the key core targets of the high frequency herb-pairs, the network of Chinese medicine-compound-target-pathway related to Dangshen were built to explore the preventing and treating molecular mechanism on gastrointestinal diseases. At last, the relation of the main active components from Dangshen and its herbal pairs with target proteins were validated by Systems Dock Web Site. The 185 formulae were selected from 543 formulae containing Dangshen, and 6 herbal pairs with Dangshen, which includes Angelica (Danggui), Licorice (Gancao), Atractylodes macrocephala (Baizhu), Poria cocos (Fuling), dried tangerine peel (Chenpi) and Astragalus membranaceus (Huangqi), were discovered with Apriori algorithm. The combination of 6 herbal pairs is similar to Bu Zhong Yi Qi Decoction; 6 herbal pairs with Dangshen were related to the target of POMC, OPRM1, CCR9 and HTR2C in TCMIP. The known targets (HTR2C, POMC, OPRM1, CCR9, OPRD1) and potential drug-targets (GNB1, GCK, SDHD, SLC25A2, DHRS4) for gastrointestinal diseases were predicted about the high frequency pairs with Dangshen. The results of GO enrichment analysis showed that the biological function was mainly located in the mitochondria and myelin sheath, and involved in the biological processes of three carboxylic acid cycle, platelet activation, and aspartic acid metabolism. KEGG pathway enrichment analysis showed that the main metabolic pathways related with Dangshen pairs involved amino acid metabolism, energy metabolism and endocrine metabolism. The prediction results showed many targets of the frequency herbal pairs with Dangshen preventing and treating gastrointestinal diseases were related with nerve cells. These herbal pairs could prevent and treat the gastrointestinal diseases through the neuroendocrine system and the brain gut axis.
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GAPDH(glyceraldehyde-3-phosphate dehydrogenase) gene is a key enzyme gene in carbohydrate metabolism and always used as reference gene. To clarify and complete the biosynthetic pathway of polysaccharide, the GAPDH gene in Codonopsis pilosula, named CpGAPDH, was cloned according to the transcriptome of pilosula, using the GAPDH gene in potato as query. The CpGAPDH contained a 1 014 bp open reading frame(ORF) and encoded a protein with 337 amino acids. Bioinformatic analysis clearly suggested that CpGAPDH shared high similarity with GAPDH among other plants, and had the closest relatives to potato and danshen. The predicted protein did not have signal peptide, which indicated that it might be located in the cytoplasm. According to the existing of several phosphorylation sites and the conserved domains analysis, we predicted that it belonged to Gp_dh_N superfamily. Prokaryotic expression showed that the recombinant expressed a 44.3 kDa protein, which was corresponding to the theoretical relative molecular mass. However, the relative transcript level of the CpGAPDH did not have significant differences in different tissues and roots at different developmental stages of pilosula. Moreover, the stability of the CpGAPDH was analyzed by BestKeeper, geNorm, and NormFinder and RefFinder software, which showed that the CpGAPDH was more stable and could be used as a new reference gene. All these lay a foundation for the expression analysis of the gene relative to the polysaccharide synthesis.
Subject(s)
Codonopsis , Genetics , Glyceraldehyde-3-Phosphate Dehydrogenases , Genetics , Plant Proteins , Genetics , Polysaccharides , TranscriptomeABSTRACT
OBJECTIVE To investigate the protective effect of Codonopsis Pilosula Polysaccharide (CPPS) on improving of the memory consolidation disorder induced by Cycloheximide and its possible mechanisms in mice. METHODS The mice was divided into five groups, as normal control group, cycloheximid model group, piracetam positive control group, CPPS 300 mg · kg- 1 group, and CPPS 150 mg·kg-1 group. The mice respectively were given saline, piracetam, and CPPS for 15 d. The memory consolidation disorder model in mice was established by ip. Cyclohexylamine, and orally administered CPPS(300 mg·kg-1 or 150 mg·kg-1) every day. Then experimental groups were subjected Morris Water Maze test. Western blotting analysis were used to analysis the expression of CaMKⅡ/CREB signaling pathways. RESULTS Morris water maze experiment showed that cyclohexylamine can cause memory consolidation disorder(P<0.01), and giving piracetam and CPPS (300 mg · kg- 1) can improve spatial memory impairment in mice(P<0.05, P<0.01). Western blotting experiment results show that compared with normal control group, CaMKⅡ and CREB contents of brain in model group mice had significant decreased(P<0.001); Compared with model group, CaMK Ⅱ and CREB contents of brain tissue in piracetam and CPPS groups increased significantly(P<0.05,P<0.01,P<0.001). CONCLUSION Cyclo?heximide can induce the memory consolidation disorder, and its effect in mice related to CaMK/CREB signaling pathways. CPPS can improved this memory disorder by influence CaMKⅡ/CREB signaling pathways.
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OBJECTIVE:To establish a method for the content determination of saccharide in polysaccharides containing galact-uronic acid based on phenol-sulfuric acid method combined with correction factor method. METHODS:The determination condi-tion of phenol-sulfuric acid was optimized. A series of standard curves were drawn with glacturonic aid-glucose mixed control with different mass ratio. The content of saccharide in Codonopsis pilosula polysaccharides CPP1b samples containing galacturonic acid was determined. According to regression equation of galacturonic acid-glucose ratio of 0-100%,the correction factor was calculated by using C. pilosula polysaccharides CPP1b as the reference polysaccharide,and the results of content determination of saccharide were corrected. The rationality of this method was verified by using mixed monosaccharide control with same composition as C. pi-losula polysaccharides CPP1b. RESULTS:The correction factor of C. pilosula polysaccharide CPP1b to glucose was 3.33;in vali-dation test,the content of saccharide in mixed monosaccharide control with same composition as C. pilosula polysaccharides CPP1b was 103.47%. RSDs of precision and stability tests was <1%. The recoveries ranged 93.52%-107.35%(RSD=5.09%,n=6). CONCLUSIONS:The established method can accurately determine the content of saccharide in C. pilosula polysaccharides CPP1b containing galacturonic acid.
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Objective: To isolate and purify the Codonopsis pilosula polysaccharide (CPP) and investigate its structure characterization and anti-oxidant activity. Methods: CPP was extracted by heating reflux, crud polysaccharide was refined and then isolated by hollow fiber ultrafiltration experimental device, the anti-oxidant activity of polysaccharides was studied by in vivo and in vitro methods. Results: CPP was purified and obtained three ingredients named CPP1, CPP2, and CPP3. The anti-oxidant activity in vitro showed that CPP3 had the strongest scavenging ability on DPPH, hydroxyl radical, and superoxide anion. In vivo study showed that high dose group of CPP3 had obvious protective effect on the mice induced by D-galactose. Conclusion: CPP has obvious anti-oxidant function. This study provides the theoretical basis for the development of CPP functional food.
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Objective: To establish an ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous determination of 23 plant growth regulator (PGR) residues in Chinese materia medica (CMM). Methods: Samples were extracted with acetontrile, and then determined by UPLC-MS/MS directly without further clean up. The matrix-matched external standard method was used for quantitative analysis. Results: The calibration curves showed good linearity in each range with correlation coefficients greater than 0.99. The limits of detection (LODs) were 0.01-20.80 ng/mL for the 23 PGRs spiked in Codonopsis pilosula and Angelica dahurica. The recoveries of the 23 PGRs spiked in C. pilosula and A. dahurica at three levels of 0.01, 0.04, and 0.1 mg/kg were in the range of 71.0%-101.4%, the relative standard deviation (RSDs) were 0.8%-15.2%. Commonly used CMM including eight species (63 batches) was analyzed by this method. Conclusion: The method proved to be simple, rapid, sensitive, accurate, and suitable for the simultaneous determination of 23 PGR residues in CMM
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Objective: To study the chemical constituents of Codonopsis pilosula. Methods: The chemical constituents were isolated and purified by AB-8 macroporous adsorption resin, silica gel, and Sephadex LH-20 column chromatography. The structures of the isolated compounds were identified on the basis of spectroscopic methods including 1H-NMR, 13C-NMR, CD, and MS spectra. Results: Seven compounds were identified as (-)-(8R, 9R, 2E, 6E, 10E)-tetradeca-2, 6, 10-triene-4-yne-8, 14-diol-9-β-D-glucopyranoside (1), phenyl-β-D-glucoside (2), (E)-isoconiferin (3), coniferin (4), syringin (5), apigenin (6), and adenosine (7). Conclusion: Compound 1 is a new compound, named lobetyolin A, and compounds 2 and 3 are all isolated from this plant for the first time.
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A new neolignan, (-)-(7R,8S,7'E)-3',4-dihydroxy-3-methoxy-8,4'-oxyneoligna-7'-ene-7,9,9'-triol(1), and seven known compounds, 9-(tetrahydropyran-2-yl)-nona-trans,trans-2,8-diene-4,6-diyn-1-ol (2), 9-(tetrahydropyran-2-yl)-trans-non-8-ene-4,6-diyn-1-ol (3), lobetyol (4), lobetyolin (5),dehydrodieoniferyl alcohol (6), 5-hydroxymethylfurfural (7), and 4, 4'-dihydroxy-3, 3'-dimethoxy-trans-stilbene (8), were isolated from the H2O extract of Codonopsis pilosula. The structures of 1-8 were elucidated by spectroscopic methods including NMR, HR-ESI-MS, and CD. In addition, compounds 2 and 3 were isolated from the genus Codonopsis for the first time.