Antioxidant and anti-inflammatory activities of four kinds of Huangshan chrysanthemum / 药学实践杂志

Objective To investigate the antioxidant and anti-inflammatory activities of four kinds of Huangshan chrysanthemum. Methods ABTS, FRAP and DPPH were used to detect the antioxidant activities of Huangshan golden silk chrysanthemum, Huangshan chrysanthemum, Huangshan gongju, and Huangshan dendranthema. Their anti-inflammatory activities were evaluated by NF-κB reporter gene assay and rat foot swelling models. Results The outcomes of ABTS，FRAP and DPPH showed that the water extracts of four kinds of chrysanthemum all had certain antioxidant activities and the activities of Huangshan golden silk chrysanthemum were strongest, followed by Huangshan chrysanthemum , Huangshan gongju , and Huangshan dendranthema. Results of NF-κB reporter gene assay and rat foot swelling models showed that four extracts of chrysanthemum morifolium could inhibit the transcription of NF-κB induced by LPS and alleviate foot swelling of rat induced by carrageenan, with the strongest activity of Huangshan chrysanthemum, followed by Huangshan golden silk chrysanthemum, Huangshan gongju, and Huangshan dendranthema. Conclusion The antioxidant activities of Huangshan golden silk chrysanthemum were strongest, followed by Huangshan chrysanthemum, Huangshan gongju, and Huangshan dendranthema. The anti-inflammatory activities of Huangshan chrysanthemum were strongest, followed by Huangshan golden silk chrysanthemum, Huangshan gongju, and Huangshan dendranthema.

Genetic diversity analysis and fingerprints of Chrysanthemum×morifolium based on SSR molecular markers / 生物工程学报

The present study aims to explore the genetic diversity of germplasm resources of Chrysanthemum×morifolium (hereinafter, C.×morifolium) at the molecular level and to establish a fingerprint database of C.×morifolium varieties. We employed 12 pairs of primers with high levels of polymorphism, clear bands, and high degrees of reproducibility to analyze the SSR molecular markers and genetic diversity of 91 C.×morifolium materials and 14 chrysanthemum- related materials. With regard to constructing the fingerprints of the tested materials, we chose 9 pairs of core primers. The findings revealed that 12 primer pairs detected 104 alleles in 105 samples, ranging from 2 to 26. The average number of observed alleles (Na) per site was 9.25. The average number of effective alleles (Ne) per site was 2.745 6, with its range being 1.276 0 to 4.742 5. Shannon genetic diversity index (I) values ranged between 0.513 3 and 2.239 9 (M=1.209 0). Nei's gene diversity index (H) ranged between 0.216 3 and 0.789 1 (M=0.578 0). The observed heterozygosity (Ho) ranged between 0.223 3 and 0.895 2 (M=0.557 5). The expected heterozygosity (He) ranged between 0.217 4 and 0.793 3 (M=0.580 8). The polymorphism information content (PIC) ranged between 0.211 5 and 0.774 0 (M=0.532 9). The genetic similarity (GS) ranged between 0.228 5 and 1.000 0 (M=0.608 3). Cluster analysis revealed that when the genetic distance (GD) equals to 0.30, the tested materials can be classified into 2 groups. When the GD equals to 0.27, the first group can be divided into 6 subgroups; accordingly, 105 tested materials can be divided into 7 subgroups. The cophenetic correlation test was carried out based on the cluster analysis, and the corresponding results showed that the cluster map correlated with the genetic similarity coefficient (r=0.952 73). According to the results of Structure population analysis, we obtained the optimal population number, with the true number of populations (K) being 3 and the population being divided concerning Q≥0.5. Three subgroups, i.e., Q1, Q2 and Q3, included 34, 33 and 28 germplasms, respectively, and the remaining 10 germplasms were identified as the mixed population. During the experiment, 9 pairs of core primers were screened among the total of 12 for a complete differentiation regarding 105 tested materials, and the fingerprints of 91 C.×morifolium materials and 14 chrysanthemum-related materials were further constructed. Overall, there were significant genetic differences and rich genetic diversity among C.×morifolium materials, which would shed light on the garden application and variety selection fields of C.×morifolium. The fingerprint database of 105 C.×morifolium varieties and chrysanthemum-related species may provide technical support for future research regarding the identification and screening system of C.×morifolium varieties.

Fusarium Wilt Changes Microbial Community Structure in Rhizosphere Soil of Chrysanthemum morifolium / 中国实验方剂学杂志

Objective:To explore the differences in rhizosphere microbial community structure between <italic>Fusarium</italic> wilt-infected and healthy <italic>Chrysanthemum morifolium </italic>plants<italic>.</italic> Method:The rhizosphere soils of diseased and healthy<italic> C. morifolium </italic>plants were sampled and subjected to high-throughput 16S ribosomal DNA （rDNA） and internal transcribed spacer （ITS） sequencing， to identify the microbial community structure including bacteria and fungi. Result:<italic>Fusarium</italic> wilt reduced the bacterial abundance and diversity but had no significant effect on fungal alpha-diversity.The proportions of Acidobacteria， Gemmatimonadetes， and Nitrospirae in rhizosphere soil of healthy <italic>C.morifolium</italic> plants were higher than those of diseased plants， while the proportions of Proteobacteria and Bacteroidetes were lower（<italic>P</italic><0.05）. <italic>Fusarium</italic> fungi accounted for 27.49%， 14.53%， and 11.94% in diseased plants whereas 0.47%， 1.01%， and 0.67% in healthy plants.Pathogenic bacteria <italic>Pectobacterium</italic> and <italic>Dickeya</italic> were enriched in rhizosphere soil of diseased plants. The abundances of nitrifying， detoxifying， and photosynthetic bacteria in rhizosphere soil of healthy plants were higher than those of diseased plants. Conclusion:<italic>Fusarium</italic> wilt reduces the bacterial richness and diversity and triggers the enrichment of massive <italic>Fusarium</italic> fungi， <italic>Pectobacterium</italic>， and <italic>Dickeya</italic>. The proportion of beneficial bacteria in rhizosphere soil of healthy plants is significantly higher than that of diseased plants.

A new polyacetylene from the stems and leaves of Chrysanthemum morifolium Ramat / 药学学报

Eight polyacetylenes were isolated from the extract of the stems and leaves of Chrysanthemum morifolium by various chromatographic methods. Their structures were determined as 2E,4E,12Z-tetradecatriene-1-pyrrolidine-1-oxo-8,10-diynoic (1), tetradeca-2E,4E,12E-trien-8,10-diynoic acid pyrrolidide (2), tetradeca-2E,4E-dien-8,10-diynoic acid pyrrolidide (3), tetradeca-2E,4E,10Z-trien-8-ynoic acid pyrrolidide (4), 2E,4E,12E-tetradecatriene-8,10-diynoic acid isobutylamide (5), 2E,4E-undecyldiene-8,10-diynoic acid isobutylamide (6), 2E,4E,10E-N-isobutyl-2,4,10-tetradecatrien-8-ynoic acid amide (7), and undeca-2E,4E-diene-8,10-diynoic acid phenylethylamide (8) by spectroscopic methods, including UV, IR, ESI-MS, HR-ESI-MS, 1D and 2D NMR spectra. Among them, compound 1 is a new polyacetylene, and compounds 2-8 were isolated from this plant for the first time. Compounds 5-8 inhibited the proliferation of A549 cell significantly at certain concentration, showing potent antitumor activity.

Correlations between content of linarin in Chrysanthemum indicum and climatic factors in habitats / 中国中药杂志

Chrysanthemi Indici Flos(CIF), the capitulum of Chrysanthemum indicum, is widely used in proprietary Chinese medicine and daily chemical products. At present, CIF is mainly produced from wild resources and rarely cultivated. This study aims to reveal the correlations between linarin content in CIF and climatic factors in different habitats, and provide a theoretical basis for suitable zoning and rational production of medicinal materials. The content of linarin in CIF was determined by HPLC. Grey relational analysis and Pearson correlation analysis were carried out for linarin content with climatic factors. The results showed that the content of linarin in CIF was significantly different among different habitats. The grey relational degrees of climatic factors with linarin content was in an order of average annual precipitation>annual average sunshine hours>annual average temperature>longitude>annual frost-free period>latitude>altitude. Longitude, annual average temperature and average annual precipitation had significantly positive correlations with the content of linarin in CIF, whereas latitude and altitude showed negative correlations with it. The annual frost-free period and annual average sunshine hours had no significant correlation with the content of linarin in CIF. The content of linarin in CIF varied significantly in different habitats. High longitude, low latitude, low altitude, high annual average temperature and high annual average precipitation could be used as indicators for the habitats of high-quality Ch. indicum. This study provides a reference for selecting suitable producing areas of Ch. indicum and establishing artificial cultivation system.

Research progress of pesticide residues in Chrysanthemum / 中国中药杂志

Chrysanthemum is widely used as a type of edible flower and also considered as the important materials of many beverages in China. Due to the occurrence of diseases and pests, and the lack of regulations for species, frequency, dose of pesticides in Chrysanthemum, pesticides have become one of the main pollutants in Chrysanthemum. The pesticide residues in Chrysanthemum were detected frequently and worth noting. This paper focused on the types of pesticides, pesticide residue detection techniques, and risk assessment methods for Chrysanthemums on the basis of relevant literatures. The pesticide residues of traditional Chinese medicine are mainly organochlorines, organophosphorus and pyrethroids, and the detection techniques include gas chromatography(GC), liquid chromatography(LC) or both combined with mass spectrometry(MS). With the increasing use of traditional Chinese medicine, Chrysanthemum is widely circulated in the market. Therefore, it is important to understand the current situation of pesticide residues in different varieties of Chrysanthemum, so as to provide theoretical reference for the control of quality and safety of Chrysanthemum and the formulation of the maximum residue limit.

Content characteristics of mineral elements and its correlation with soil nutrients and effective components in Chrysanthemum morifolium from different producing areas in Macheng / 中国中药杂志

In this study, 24 copies of samples of Chrysanthemum morifolium and soil from two main production towns in Macheng city were collected, and the contents of 13 mineral elements, 5 effective components and 14 soil nutrient factors in Ch. morifolium were determined. The enrichment characteristics of available soil nutrients by mineral elements were analyzed and the dominant factors affecting the effective components of Ch. morifolium were screened. The results showed that the content of mineral elements and soil nutrients and effective components are very different, and variation of soil fertility was much greater than that of inorganic elements in chrysanthemum plants. In general, the level of element content in Ch. morifolium from different producing areas is K>N>P>Mg>Ca>Fe>Mn>Zn>Cu>Ni>Cr>Pb>Cd. The content of K, N and Mg is higher than that of common crops, and the content of Cu, Cd and Pb in Ch. morifolium from various producing areas does not exceed the relevant standards. The N, P and K enrichment capacity in soil was stronger than that of other elements, and the Ca enrichment ability was the worst. The content of AvCu in the soil was positively correlated with the contents of N, Mg, K, Fe and Cu elements in Ch. morifolium. The contents of chlorogenic acid, luteolin, 3,5-O-dicaffeoylquinic acid reached the pharmacopoeia standard. The percentage of chlorogenic acid and 3,5-O-dicaffeoylquinic acid in Ch. morifolium that from Huangtugang town in the active components were generally higher than that from Futianhe town, and the diffe-rences of luteolin contents in the two producing areas were relatively small. The correlation and regression analysis showed that the contents of Cu, Zn and Cr in Ch. morifolium were positively correlated with the active components, while the contents of Fe, Mn and Ni were negatively correlated with the contents of AvP, AvK, TK, AvMn and AvCu in soil. In general, Zn and Ca fertilizer should be added to the ecological planting of Ch. morifolium, K fertilizer should be added, and N and P fertilizer should be applied appropriately.

Analysis and evaluation of mineral elements of Chrysanthemum morifolium for medicinal and tea use of different germplasm resources / 中国中药杂志

In this study, 23 germplasm resources of Chrysanthemum morifolium used in medicine and tea were collected from Dabie Mountains and its surrounding producing areas, and the contents of 13 mineral elements were determined and compared. The thermal maps of correlation analysis, principal component analysis and cluster analysis were used for comprehensive evaluation. The results showed that the average content of each element in Ch. morifolium of different germplasm resources was: K>N>P>Mg>Ca>Fe>Mn>Zn>Cu>Ni>Cr>Pb>Cd, and the leaves were: K>N>Ca>Mg>P>Fe>Mn>Zn>Cr>Cu>Ni>Pb>Cd. There are rich contents of N, P, K, Ca, Mg and Fe in Ch. morifolium flowers and their leaves, among them, K element has the largest change range, while N, Ca, Fe, Mg and Zn elements have a larger change range. The absorption and accumulation of each element in the leaves of different germplasm resources varied greatly. The correlation analysis shows that there is a strong positive correlation between Ca element, Mg, Mn and Cd element.Principal component analysis in Ch. morifolium flowers characteristic elements for Mn, Cr, Cu, P, K, can be used as a Ch. morifolium resources to identify the characteristics of the elements, choose top five principal component(F1-F5) comprehensive evalua-tion of medicinal Ch. morifolium, scored in the top five varieties for Hangiu-Fuhuangju, Hangju-Xiaoyangju, Hangju-Sheyangju, Hangju-Dayanghua, Hangju-Subeiju,indicates that in terms of mineral elements, the five medicinal Ch. morifolium resources quality is better. The PCA score chart can divide 23 Ch. morifolium resources into 4 groups, and the cluster analysis heat map divides 23 Ch. morifolium resources into 5 groups. All the Ch. morifolium resources of the same type can be well clustered together, indicating that the difference in mineral element content of Ch. morifolium germplasm resources is closely related to genetic factors.

Comprehensive antioxidant and anti-inflammatory activity of alcohol extracts from Chrysanthemum indicum in different areas based on entropy weight and TOPSIS methodology / 中国中药杂志

The study is aiming at investigating the application of entropy weight TOPSIS method in the comparison of the scavenging effect of DPPH, ABTS and hydroxyl radical and the inhibition effect of xanthine oxidase(XOD) and lipoxygenase(LOX) of Chrysanthemum indicum. The DPPH, ABTS, salicylic acid and spectrophotometry were used to determine the scavenging effect of DPPH, ABTS and hydroxyl radical and the inhibition effect of xanthine oxidase(XOD) and lipoxygenase(LOX) of Ch. indicum from 31 different areas in vitro. Take the half inhibition rate of as the evaluation index, two principal components were extracted by the principal component analysis, and their cumulative contribution rate reached at 92.4%. The different areas of Ch. indicum could be divided into Dabei Mountain and Qinling-Taihang Mountain by use principal component to analysis. The entropy weight TOPSIS method was used to objectively assign weights to five indexes, calculate the weight of each index and set up the best and worst scheme of the evaluation object, and the relative proximity(C_i) was used as the measure to construct the multi-index comprehensive evaluation model of Ch. indicum. And then sort with the relative proximity value. The results showed that the relative proximity was between 0.098 and 0.983 which represents there were significant differences in the scavenging effect of DPPH, ABTS and hydroxyl radical and the inhibition effect of xanthine oxidase(XOD) and lipoxygenase(LOX) between extracts of Ch. indicum from different areas. The Ch. indicum from Dabie Mountain area have a relatively high relative degree of measurement and high-quality ranking. Taken together, the quality of Ch. indicum.from the Dabie Mountain area is better. The index weight coefficient and the classification result of producing area are basically consistent with the result of principal component analysis. The results show that the TOPSIS method based on entropy weight method can be used to evaluate the comprehensive quality of Ch. indicum.

Ectopic expression of Chrysanthemum CDM19 in Arabidopsis reveals a novel function in carpel development

BACKGROUND: APETALA3 (AP3) has significant roles in petal and stamen development in accordance with the classical ABC model. RESULTS: The AP3 homolog, CDM19, from Chrysanthemum morifolium cv. Jinba was cloned and sequenced. Sequence and phylogenetic analyses revealed that CDM19 is of DEF/AP3 lineage possessing the characteristic MIKC-type II structure. Expression analysis showed that CDM19 was transcribed in petals and stamens of ray and disc florets with weak expression in the carpels. Ectopic expression of CDM19 in Arabidopsis wild-type background altered carpel development resulting in multi-carpel siliques. CDM19 could only partially rescue the Arabidopsis ap3­­3 mutant. CONCLUSIONS: Our results suggest that CDM19 may partially be involved in petal and stamen development in addition to having novel function in carpel development.

Effects of planting density on yield and quality of Chrysanthemum morifolium / 中国中药杂志

In this paper, five field density treatments were set up in the field plot experiment, which were 2 500,3 000,5 000,6 660,8 000 plants/mu(1 mu≈667 m~2). The agronomic traits, economic traits, mineral element absorption and the content of effective components of Chrysanthemum morifolium under different densities were studied. The results showed that dense planting could significantly reduce the number of secondary branches of Ch. morifolium and the yield per plant, but significantly increase the population yield of Ch. morifolium. The yield of Ch. morifolium was the highest when the density was 8 000 plants/mu, but the effect of increasing yield would gradually decrease with the increase of planting density. With the increase of planting density, the N, P and Mg elements in flowers firstly increased and then decreased. The N element content in leaves increased gradually, which showed that increasing the planting density within a certain range could increase the absorption of N, P and Mg elements in flowers and leaves of Ch. morifolium. The contents of rutin, chlorogenic acid and 3,5-O-dicaffeoyl quinic acid in Ch. morifolium showed a trend of first increasing and then decreasing with the increase of planting density. When the planting density was 5 500,5 000,3 750 plants/mu, the content of chlorogenic acid, rutin and 3,5-O-dicaffeyl quinic acid had the maximum value. The content of luteolin in Ch. morifolium decreased gradually with the increase of planting density. When the planting density was 7 143 plants/mu, the content of luteolin was the minimum. Considering factors such as yield and active ingredient content, the cultivation density of 5 000 plants/mu(row spacing 40 cm×30 cm) can be selected for standard planting of Ch. morifolium.

Status and prospects of resource utilization of non-medicinal parts of medicinal Chrysanthemum morifolium / 中草药

Chrysanthemum morifolium is a traditional Chinese medicinal material in China. The yield of non-medicinal parts is much higher than the inflorescence, and the yield of stems and leaves of C. morifolium is 3.5 times of medicinal parts. For a long time, the non-medicinal parts of C. morifolium have not been fully used, resulting in great waste of resources and environmental pollution. Therefore, the in-depth research and development of non-medicinal parts of C. morifolium deserve attention. Research shows that the non-medicinal parts of C. morifolium is rich in volatile oil, flavonoids, phenolic acids, polysaccharides and other components, which have antibacterial, anti-inflammatory, antioxidant, anti-convulsion and improvement of intestinal disorders. This article summarizes the research situation of chemical components, pharmacological effects, and resource utilization status of stems, leaves, roots and other non-medicinal parts produced during the cultivation and production of medicinal C. morifolium, in order to provide the scientific basis and reference for the development, utilization and industrialization of the non-medicinal parts of medicinal C. morifolium.

Quality Evaluation of Chrysanthemum indicum Based on Fingerprint, Pattern Recognition and Simultaneous Determination of Multi-Component / 中国药学杂志

OBJECTIVE: To establish a method based on HPLC fingerprint, pattern recognition and simultaneous determination of multi-component for quality evaluation of Chrysanthemum indicum, and investigate its feasibility and accuracy. METHODS: HPLC fingerprints of 21 batches of Chrysanthemum indicum were established, similarity analysis, principal component analysis and cluster analysis were performed on fingerprints of Chrysanthemum indicum by TCMYS fingerprint software, and 12 common chromatographic peaks in fingerprints were identified, and 12 components were quantified by external standard method. RESULTS: The HPLC fingerprints of 21 batches of Chrysanthemum indicum were similar, and their classification trend was found. Seventeen common peaks were found and 12 of them were identified as neochlorogenic acid, chlorogenic acid, caffeic acid, 1,3-dicaffeoylquinic acid, cynaroside, isochlorogenic acid C, isochlorogenic acid A, (1S, 3R, 4R, 5R)-3, 4-bis[[(E)-3-(3, 4-dihydroxyphenyl)prop-2-enoyl]oxy]-1, 5-dihydroxycyclohexane-1-carboxylic acid, linarin, luteolin, apigenin and acacetin. Good linear relationship (r≥0.999 6) was shown for the external standard method, the average recoveries were between 95.84% and 102.46%, and the RSDs were all below 2.72%. CONCLUSION: Combination of HPLC fingerprint, pattern recognition and simultaneous determination of multi-component are accurate and reliable for quality evaluation of Chrysanthemum indicum, which can provide important information for the overall quality evaluation of Chrysanthemum indicum.

Effects of planting period on growth, yield and medicinal quality of Chrysanthemum morifolium / 中国中药杂志

In this paper, Chrysanthemum morifolium was used as the experimental object, eight different planting periods were set up in field plot experiment from April to August, which were 04-15, 05-19, 05-30, 06-09, 06-19, 07-20, 07-31, 08-15. The effects of different treatments on the occurrence of root rot, agronnmic traits, mineral element absorption and content of effective components of Ch. morifolium in Macheng country of Hubei province were studied. The results showed that delaying the planting time could effectively reduce the diseases occurrence of root rot of Ch. morifolium. With the advance of transplanting period, the plant height, the weight of one hundred flowers and the number of flowers of Ch. morifolium showed a trend of gradual decrease, while the number of primary branches and the thickness of main stem and the primary branch increased first and then decreased. The yield of Ch. morifolium per plant and per mu increased with the advance of the planting period, and the yield per mu increased during the planting period on June 19, which was 91.96% higher than that on April 15. And with the delay of the planting period,the absorption and accumulation of potassium(K) elements was promotes. The content of active ingredients such as chlorogenic acid, rutin, luteolin, and 3,5-O dicoffeoacy lquinic acid in the Ch. morifolium increased significantly and then gradually decreased with the delay of the planting period, which indicates that late planting can significantly improve the quality of Ch. morifolium. Considering factors such as the occurrence of root rot disease, yield and active ingredient content, combined with climatic conditions in the Dabie Mountains in eastern Hubei, the optimum planting period of Ch. morifolium was determined from mid-late June to early July.

Quality assessment of soil fertility of planting Chrysanthemum morifolium areas of Macheng city based on minimum data set / 中国中药杂志

A minimal data set( MDS) for soil fertility evaluation of Chrysanthemum plantation areas of Macheng city was established by principal component analysis( PCA) combined with Norm values of soil fertility indices and correlation coefficients among indices. A radar map was used to visually reflect the fertility level of individual indicators. Then,the comprehensive index model was used to calculate the soil fertility quality index( SFQI),and the values of SFQI was used to cluster,and the results showed that MDS was composed of five indicators: organic matter( OM),total phosphate( TP),available phosphorus( Av P),available magnesium( Av Mg) and available ferrum( Av Fe). Radar maps showed that the fertility of available phosphorus( Av P) and available copper( Av Cu) was mostly different in the two town,and the fertility of available ferrum( Av Fe) is smallest different. Except for the effective manganese( Av Mn) fertility level of Huangtugang town was higher than that of Futianhe town,the rest were lower than that of Futianhe town. Through analysis,the sensitivity of SFQI value calculated by taking the contribution rate of MDS index in the principal component of the whole data set( TDS) as the weight was the highest,MDS could better replace TDS. The value of SFQI-MDS ranged from 0. 353 to 0. 833,with an average value of 0. 604 and a coefficient of variation of 22%. The results of SFQI-MDS clustering showed that soil fertility could be divided into four categories: grade Ⅰ( 0. 727-0. 833) was superior,accounting for 25. 0%,grade Ⅱ( 0. 615-0. 681)was good,accounting for 29. 2%,mainly distributed in Futianhe Town,grade Ⅲ( 0. 494-0. 589) was medium,accounting for29. 1%,and grade Ⅳ( 0. 353-0. 419) was poor,accounting for 16. 7%,mainly distributed in Huangtugang town. Soil fertility of Futianhe town was better than that of Huangtugang town. It is suggested that boron fertilizer and potassium fertilizer should be supplemented to Chrysanthemum morifolium in production practice,and the amount of phosphate fertilizer,magnesium fertilizer and nitrogen fertilizer should be increased appropriately. At the same time,the amount of organic fertilizer should be increased to enhance soil fertility and improve soil physical and chemical properties.

Effect of different sulfur fumigation dosages on activity of browning enzymes and chemical constituents of Chrysanthemum morifolium cv. Boju / 中国中药杂志

This study aims to investigate the influence of the different dosages of sulfur on the quality and the browning enzyme activity of Chrysanthemum morifolium cv. Boju. In this experiment,UV-spectrophotometry was used to determine the activities of browning enzymes,including polyphenol oxidase( PPO) and peroxidase( POD),in 7 different dosages of 0,4,8,16,50,150,200 g·kg~(-1)( weight ratio of sulfur/fresh chrysanthemum). A comprehensively comparison of the 7 chemical constituents of C. morifolium cv. Boju fumigated with 7 different dosage of sulfur was conducted by HPLC analysis. In this paper,the results showed that the activities of PPO and POD enzymes decreased significantly in chrysanthemum processed by sulfur fumigation. The activities of PPO and POD enzymes decreased gradually with the increase of sulfur dosage. When the sulfur dosage was higher than 4 g·kg~(-1),the PPO enzyme was significantly reduced. When the sulfur dosage was higher than 8 g·kg~(-1),the PPO enzyme was completely inactivated. The effect of different sulfur dosage s on the chemical composition was investigated. In comparison,it was found that when the sulfur dosage was 8 g·kg~(-1),the content of chlorogenic acid was higher than the 4 g·kg~(-1) and that of the sample without sulfur fumigation. Thereafter,with the increase of the sulfur dosage,the content of chlorogenic acid was unchanged. It was speculated that when harvesting,the tissue of fresh flower was destroyed,which caused the activation of browning enzymes. Afterwards,the sulfur fumigation could significantly reduce the activity of browning enzymes,which prevented the conversion of phenols in the reaction substrates( chlorogenic acid and 3,5-dicaffeoylquninic acid) into terpenoids,and better retained quinic acid components. However,when the sulfur dosage reached 8 g·kg~(-1) or16 g·kg~(-1),the content of quinic acid components were no longer changed,which indicated that the sulfur dosage had reached the saturated dosage. Similarly,when the sulfur dosage was increased,the contents of flavonoid aglycones showed a downward trend except for luteolin-7-O-glucoside. It was speculated that the sulfur fumigation inhibited the activity of hydrolase,which reduced the hydrolysis of flavonoid glycosides to aglycones. However,the reaction mechanism needed further verification. In conclusion,although sulfur fumigation could significantly inhibit browning,different dosages of sulfur had a significant effect on the chemical composition of C. morifolium cv. Boju,which could affect the consistency of quality and the stability of the therapeutic effect. Excessive use of sulfur was likely to cause a large amount of SO2 residues in C. morifolium cv. Boju,Therefore,different Sulphur dosages had a significant effect on the quality of chrysanthemum,which therefore was not recommended in production. A small dose of sulfur could be used to prevent enzymatic browning. When the dosage of sulfur increased to a certain extent or reached a saturation state,a small dose of sulfur is recommended in necessary. In this paper,the correlation between the sulfur dosage,the enzyme activity,and the main chemical constituents of chrysanthemum was clarified. The experimental research provided the guidance for regulating the harvesting processing of chrysanthemum and the harvesting processing,and improving the quality of chrysanthemum.

Application of ITS2 secondary structure phylogenetic information in DNA barcode identification of Chrysanthemum indicum and its related plants / 中国中药杂志

By exploring additional phylogenetic information hidden in ITS2 secondary structure,the possibility of identifying Chrysanthemum indicum and its related species with DNA barcode of ITS2 nucleic acid sequence and its structure information were discussed.The genomic DNA was extracted from 12 samples. The ITS2 fragments were amplified by PCR and sequenced bidirectionally to obtain ITS2 sequence information. 28 sequences of related species for Ch. indicum were downloaded from Gen Bank. Until all 40 ITS2 sequences were aligned,ITS2 secondary structure prediction and structure comparison were finished. Then ITS2 secondary structure information was coded. After comparing ITS2 structure information and nucleic acid information,MP phylogenetic trees were built. The results showed that the secondary structures of ITS2 shared the same structure model--a four-fingered hand. They not only have the common characteristics of ITS2 secondary structures in plants,but also have many other conservative sequences,and their overall conservativeness is high. Among all species used in this study,their ITS2 secondary structures had obvious difference. In addition,the number of mutation sites in the joint matrix compared with the nucleic acid sequences increased by nearly 90%,which greatly enriched the number of mutation sites. This method of information analysis distinguished Ch. indicum from its related species. At the same time,the support rate of the branches of evolutionary trees and the identification rate of species were significantly improved. Although there was no distinction between Ch. zawadskii and Ch. morifolium,it effectively distinguished the three species,namely,Ch. hypargyrum,Ch.oreastrum,and Ch. dichrum. Therefore,the authors suggest that the ITS2 sequence combined with its structural data information should be applied to the identification of Ch. indicum and its related species,and be widely applied to DNA barcode research.

Qualitative and quantitative analyses of Chrysanthemum morifolium ‘Fubaiju’ by LC-MSn and HPLC-UV / 中国药科大学学报

@#A qualitative analytical method of liquid chromatography coupled with mass spectrometry(LC-MSn)was developed for the identification of main constituents in Chrysanthemum morifolium ‘Fubaiju’. High-performance liquid chromatography(HPLC)was developed for the quantification of five active components, including chlorogenic acid(1), luteolin-7-O-β-D-glucopyranoside(2), luteolin-7-O-β-D-glucopyranuronide(3), 3, 5-Di-caffeoylquinic acid(4), and apigenin-7-O-β-D-glucopyranoside(5). A total of 22 compounds, including 13 flavonoids and 9 phenolic acids, were identified based on their retention behaviors, UV profiles and MS fragment information. Furthermore, a validation method with good linearity(r> 0. 999 9), precision, stability, repeatability and recovery was successfully applied for simultaneous determination of five major components in 10 batches of C. morifolium ‘Fubaiju’ by HPLC-UV method. The established method was proved to be a validation strategy for the quality evaluation of C. morifolium ‘Fubaiju’.

Establishment of HPLC fingerprint and comparison of active ingredients between varieties of chrysanthemum in Hubei province / 中国中药杂志

In this paper,the fingerprint of different varieties of chrysanthemum were established with " Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica" and the content of chlorogenic acid,galuteolin and 3,5-O-dicaffeoylquinic acid in 29 batches of different varieties of chrysanthemum in Futianhe town,Huangtugang town and Wuhan city were compared. At the same time,similarity evaluation and common peak clustering analysis were carried out. There were 11 common peaks in the fingerprints of 29 batches of different varieties of chrysanthemum,and the similarity ranged from 0. 802 to 0. 975. Hangju and Gongju were divided into one group by cluster analysis,and Huangju into another category. The established fingerprint method provides a basis for the identification of chrysanthemum cultivars. The content of 29 batches of chlorogenic acid was between 4. 092 and 11. 723 mg·g-1,luteolin was between 1. 010 and 11. 713 mg·g-1,and 3,5-O-dicaffeoylquinic acid was between 8. 828 and 33. 435 mg·g-1,both reach the pharmacopoeia standard,but the effective components of different varieties of chrysanthemum were quite different. Based on the contents of three active ingredients and the diversity of fingerprint peaks,the quality of the characteristic germplasm resource of local Fubaijuin Macheng is superior,and the protection of local characteristic germplasm resource should be strengthened in production.

Cloning and prokaryotic expression of CHI in Chrysanthemum morifolium cv.'Hangju' / 中国中药杂志

Three Chrysanthemum-chalcone-isomerase genes( CmCHI) were successfully cloned by PCR from the database of Chrysanthemum transcriptome and named CmCHI1,CmCHI2 and CmCHI3,respectively. Bioinformatics analysis showed that the base numbers of CmCHI1-3 open reading frame were 708,633 and 681 bp,encoding 235,210 and 226 amino acids,respectively. Three fusion proteins of about 30 kDa were successfully induced by prokaryotic expression technology,and the corresponding recombinant fusion proteins were isolated and purified by Ni-NTA resin column. Clustering analysis showed that the 3 CmCHI were homologous with Compositae plants,and CmCHI1 and CmCHI3 belonged to type Ⅰ CHI. CmCHI2 belongs to type Ⅳ CHI. Using β-actin as an internal reference gene,RT-qPCR was used to detect and analyze the expression of CmCHI1-3 genes in Hangju. The results showed that the expression levels of CmCHI1 and CmCHI3 were higher,while the expression levels of CmCHI2 were lower. It was concluded that CmCHI1 and CmCHI3 were the main chalcone isomerase genes involved in the synthesis of flavonoids in Hangju,and CmCHI2 was a helper gene. Flooding treatment significantly promoted the expression of CmCHI1 and CmCHI3 genes,but had no regulatory effect on CmCHI2. The above results provided a basis for further study of the molecular regulation mechanism of CHI gene in the metabolism of flavonoids in Hangju,which laid a foundation for improving the content of flavonoids in Hangju and finally improving the medicinal quality of Hangju.