| Chrysanthemum is a dried flower head of Chrysanthemum morifolium Ramat,which was first recorded in Shennong materia classic with a history of more than 2,000 years.Chrysanthemum can be used as medicine and food,so it is in high demand.Chrysanthemum has the effect of reducing inflammation,smoothing liver,brightening eyes,abatementing heat and detoxifying.According to different places of origin and processing methods,chrysanthemum in 2015 edition of Chinese Pharmacopoeia can be divided into five varieties: Hangju,Boju,Gongju,Chuju and Huaiju.There are also local varieties in various producing areas,such as Qiju,Jinsi huangju,Wuyuan Huangju.There has been no systematic comparative study on the consistency of agronomic characters,chemical constituents and pharmacological activities among different Chrysanthemum varieties.In recent years,in order to solve the problems of continuous cropping obstacle,diseases and insect pests,many new varieties of Chrysanthemum have been introduced or cultivated among different producing areas,which leads to the confusion of market.Therefore,it is necessary to evaluate the morphological characteristics,chemical constituents and pharmacological activities of the existing chrysanthemum varieties,which is helpful for the protection and rational utilization of chrysanthemum resources.Objective: To compare and analyze the germplasm resources of Chrysanthemum from the aspects of agronomic characters,mineral elements,effective components and pharmacological activity,so as to provide references for genetic improvement,germplasm resource protection,introduction and cultivation,quality evaluation and clinical medication.Methods:1.29 chrysanthemum resources were collected from Dabie Mountain Area and relevant origin.51 morphological characters of plants,leaves and inflorescences were investigated and analyzed.Principal component analysis and cluster analysis were used to classify them.2.The contents of 13 mineral elements in flower and leaf were determined by many ways,such as atomic absorption spectrometry,including N,P,K,Ca,Mg,Fe,Zn,Mn,Ni,Cu,Pb,Cr and Cd.The distribution law of elements was found and classified by correlation analysis,principal component analysis and cluster analysis.3.HPLC fingerprint of chrysanthemum was established and the contents of chlorogenic acid,luteolin,3,5-o-dicaffeoylquinic acid was determined for evaluating the quality of chrysanthemum resources.4.The antioxidant activities of water extracts from chrysanthemum were evaluated in vitro by DPPH,ABTS and FRAP.The protective effects of water extracts from chrysanthemum on human normal hepatocytes(L02 cells)damaged by hydrogen peroxide(H2O2)and mice with alcoholic liver injury were studied.The contents of luteolin,luteolin-7-o-?-d-glucuronide,isochlorogenic acid A,isochlorogenic acid B,isochlorogenic acid C,vanillin-7-o-glucoside,monkside and artichoke glycoside in the water extract were measured by HPLC,The correlation between chemical component and agronomic character,mineral element and antioxidant activity was analyzed.Results:1.According to the investigation of agronomic characters,34 of 51 morphological characters have the coefficient of variation more than 20%,and 6 characters more than 40%,which indicates that there are abundant genetic diversity in chrysanthemum resources.A total of 11 principal components were extracted by principal component analysis,and the cumulative contribution rate reached 89.54%.According to the score of the first three principal components,Hangju series,QYB,and CJ can be divided into a class,because diameter of tubular flowers group and number of tubular flowers are similar.DHJ?XHJ?BJ and HAJ can be divided into a class,because number of tubular flowers is few and ligulate flowers are abundant.GJ?HJ?TCJ and XNJ can be divided into a class,because the tubular flowers are petal-like,and the type of flower center is anemone.Cluster analysis of 11 principal components can better distinguish Hangju series and other varieties,which is basically consistent with the results of field investigation and statistics,indicating that the cluster method is scientific and reliable for the comprehensive classification of Chrysanthemum.2.The content of mineral element N,P,K,Ca,Mg,Fe,Zn in chrysanthemum resources is rich.According to the principal component analysis,the representative elements of chrysanthemum are Mn,Cr,Cu,P and K.according to the scatter score chart,23 chrysanthemum resources are divided into 4 groups.The early varieties of Hangju can be divided with other varieties of Hangju.According to the cluster analysis,23 chrysanthemum resources are divided into 5 groups,among which HJ-FBJ is a single group.3.The HPLC fingerprint and the determination of three effective components showed that: the difference of chromatographic peaks was mainly concentrated after 45 minutes;the similarity between the sample map and the control map was between 0.753 and 0.960,and JSH was the lowest,while HJ-DYH,HJ-EYH,HJ-ZYH,HJ-FBJ,HJ-FHJ were higher.There are 12 common peaks identified by the fingerprint.The cluster analysis shows that HJ-XYJB and HJ-XYJH are grouped into one group,BJ,QJ and HJ-BJJ are grouped into one group,JSH is divided into a group,and HAJ is divided into a group,and other varieties are grouped into one group.The contents of chlorogenic acid,luteolin,3,5-o-dicaffeoylquinic acid and 3,5-o-dicaffeoylquinic acid were 0.36% ~ 2.69%,0.02% ~ 0.63% and 0.31% ~ 4.00% respectively.The content of chlorogenic acid in HJ-BJJ and BJ is the highest,greater than 2.30%,more than 10 times higher than the standard of Chinese Pharmacopoeia.The content of luteolin in DHJ and XHJ is the highest,greater than 0.60%,more than 7.5 times higher than the standard of Chinese Pharmacopoeia.The content of 3,5-o-dicaffeoylquinic acid was 3.49% and 4.00% in BJ and HJ-BJJ respectively,which was more than 3 times higher than the standard of Chinese Pharmacopoeia.4.Pharmacological activities research found that:(1)The foud of comparative study of antioxidant activity in vitro show that the IC50 values of ·DPPH free radicals of HJ-DYH?BJ?DHJ and XHJ were 0.028 mg·m L-1?0.029 mg·m L-1?0.030 mg·m L-1 and 0.031 mg·m L-1,respectively,which showed a strong ability to scavenge ·DPPH free radicals.The IC50 values of ·ABTS+ free radicals in BJ,DHJ and XHJ were 0.026 mg·m L-1?0.025 mg·m L-1 and 0.026 mg·m L-1,respectively,which showed a strong ability to scavenge ·ABTS+ free radicals.When the concentration of water extract is 0.8 mg·m L-1,the total antioxidant capacity of XHJ is the highest,which is 0.789 m M,followed by HJ-DYH and DHJ,which are 0.693 m M and 0.680 m M respectively.(2)HJ-XYJH,HJ-TXZS,HJ-TXWS,HJ-BJJ,HJ-FHJ,TCJ,CJ,HSG,HJ-EYH and HAJ have obvious protective effects on liver injury cells,and TCJ has the strongest anti hydrogen peroxide effect on liver injury cells.(3)The protective effects of 8 kinds of chrysanthemum water extract on alcoholic liver injury mice showed that the water extract of chrysanthemum can reduce the liver coefficient and AST content of liver tissue,improve the morphology and pathological changes of liver,which shows that the water extract of chrysanthemum has protective effect on alcohol liver injury.HJ-FBJ can significantly reduce the content of AST and MDA in serum,BJ can significantly reduce the content of AST and increase the content of GSH in serum,DHJ can significantly reduce the content of ALT in liver tissue,JSH,DHJ,HSG and HAJ can significantly increase the content of GSH in liver tissue,HJ-FBJ,JSH and CJ can significantly reduce the content of MDA in liver tissue Content,HJ-FBJ,JSH,DHJ,HSG can significantly increase the content of SOD in liver tissue.Therefore,HJ-FBJ,JSH,DHJ and BJ have stronger protective effects on alcoholic liver injury mice.5.The contents of active components of water extract from chrysanthemum and their correlation with agronomic characters,mineral elements and antioxidant activity showed that:(1)the contents of chlorogenic acid,isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C in BJ were higher than those of other varieties,and the contents of luteolin and luteolin-7-o-?-d-glucuronide in of DHJ and XHJ were higher than those of other varieties.The content of geranioxylin 7-o-glucoside was higher in early-maturing Hangju variety.(2)Chlorogenic acid,isochlorogenic acid B,A,C and luteolin are mainly found in the corolla tube of glossy floret,and most of them are in the corolla tube of glossy floret.There is a very significant negative correlation between the content of luteolin-7-o-glucoside and the florescence,that is,the later the florescence,the lower the content of geranioxylin-7-o-glucoside.Geranioxylin-7-o-glucoside and buddleoside are mainly in the tube flower.(3)There was a significant positive correlation between isochlorogenic acid A and tilianin,N and P,and a significant positive correlation between isochlorogenic acid B and n,Chlorogenic acid,isochlorogenic acid A,C and luteolin had significant negative correlation with Pb,luteolin had significant negative correlation with Mn, geranioxylin-7-o-glucoside had significant positive correlation with Mn and Mg,and had significant positive correlation with Ca and Pb,while buddleoside had significant positive correlation with Cd and Cr.(4)The contents of chlorogenic acid,luteolin,isochlorogenic acid A,isochlorogenic acid B,isochlorogenic acid C,luteolin-7-o-?-d-glucuronide are positively correlated with ·DPPH free radical scavenging rate,· ABTS + free radical scavenging rate and total antioxidant capacity,which are the effective components of antioxidant.There was a negative correlation between the three antioxidant indexes and the survival rate of hepatocytes injured by hydrogen peroxide.The anti-hydrogen peroxide liver cell damage were negatively correlated with the contents of chlorogenic acid,luteolin-7-o-?-d-glucuronide,isochlorogenic acid A,isochlorogenic acid B,isochlorogenic acid C and tilianin,but positively correlated with the contents of vanillin-7-o-glucoside and monkside,but not significantly.Conclusion: Different chrysanthemum germplasm resources have great differences in agronomic characters,active ingredients,mineral elements and antioxidant activities.There are abundant genetic diversity and excellent characters in different chrysanthemum germplasm resources,which lay a good foundation for the breeding of chrysanthemum varieties for medicine and tea and the protection of new varieties.The results of this study can provide reference for genetic improvement,germplasm resource protection,introduction and cultivation,quality evaluation and clinical use of chrysanthemum germplasm resources. |
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