| Purpose:Liaoning Province is a real estate area for gentian medicine,but due to the destruction of vegetation and over-excavation by humans,wild gentian resources are very scarce.In this experiment,wild and cultivated gentian medicinal materials from different places in Liaoning Province were used as research objects.The TOE method was used to comprehensively compare the contents and growth indices of eight chemical components of wild and cultivated gentian.From a macro perspective to a micro perspective,a comprehensive comparison of the quality of gentian herbs can provide theoretical basis for the cultivation of gentian to replace wild gentian herbs,and provide scientific guidance for the cultivation of gentian herbs.Materials and methods : 1 materials: The experimental materials of this subject were collected in 9 different wild or cultivated producing areas in Liaoning Province and gentian grasses of different bases in various producing areas.(Qingyuan County,Fushun City,Liaoning Province;Xinbin County,Fushun City,Huanren County,Benxi City),medicinal parts were its root and rhizome.The materials were identified as Gentiana scabra Bunge.By professor Yin Haibo of Liaoning University of Traditional Chinese Medicine.Gentiana seed and 2-year-old Seedling collected in Ying'e Men Zhen Ying'e Men Cun,Qingyuan County,Fushun City which all varieties are Gentiana scabra Bunge.The experimental soil samples were collected for the cultivation base corresponding to the 14 batches of medicinal materials.2 Method :2.1 Straightedge,vernier caliper direct measurement and color difference measuring method were used to determine the growth index and medicinal powder color,and the medicinal materials were photographed and stored.From the perspective of appearance characteristics,the differences between wild and cultivated gentian in different producing areas were compared.2.2 The contents of flavonoids and polysaccharides in gentian medicinal materials were determined by ultraviolet spectrophotometry,and the results were analyzed by significant difference test(LSD method).2.3 High performance liquid chromatography was used to determine the cyclic cyclic cyclic ether terpenoids in gentian herbs from different wild and cultivated areas,namely gentiopicroside,manganin,argentin,argentin,and apatin.Comprehensive evaluation of the quality of gentian herbs.2.4 Fingerprinting of Wild and Cultivated Gentians from Different Places by High Performance Liquid Chromatography2.5 High-throughput sequencing was used to determine the entire chloroplast genome of gentiana scabra.Results:1.Appearance traits The results of independent sample t test on the average root thickness,average whisker number,L,a,b,and E values that meet the normal distribution are as follows: average root thickness P = 0.427 > 0.05,The difference in average root thickness is not statistically significant;the average number of whiskers P = 0.074 > 0.05,the difference between the average number of roots of wild products and cultivated products is not statistically significant;L value P = 0.011<0.05;The L values of cultivated products are different;a value P = 0.62 > 0.05,the difference in a value between wild products and cultivated products is not statistically significant;b value P = 0.06 > 0.05,the difference in b value between wild products and cultivated products is not available;Statistical significance;E value P = 0.01<0.05,it can be considered that the E value of wild products and cultivated gentian is significantly different,which has statistical significance.2.Chemical composition2.1 Polysaccharides: The statistics of normality test of polysaccharide content in wild and cultivated products are 0.914 and 0.870,respectively,and the P values are 0.343 and 0.065,both of which are> 0.05,which meet the normal distribution;the four Levene statistics are 4.461 and 4.073,respectively 4.073,4.517,all> 0.05,homogeneity of variance.Independent sample t test P = 0.318 > 0.05,there was no significant difference in polysaccharide content between wild and cultivated products.2.2 Five kinds of cyclic enoterpenoid glycosides: Except for gentisin and gentioside,there are no significant differences between gentian glucoside,argentin and argentin in wild and cultivated gentian.Among them,gentisic acid is significantly related to the L value of gentian medicinal materials.It can be speculated that gentamic acid will make gentian medicinal materials whiter.2.3 Flavonoids: The statistics of the normality test of flavonoid content in wild and cultivated products are 0.825 and 0.932,and the P values are 0.282 and 0.087,all of which are> 0.05,which meet the normal distribution.4.073,3.986,both> 0.05,homogeneity of variance.Independent sample t test P = 0.031 <0.05.The difference in flavonoid content between wild and cultivated products is statistically significant.2.4 Oleanolic acid: As the wild and cultivated products do not reach the detection limit in part,the sample size is small,and there is no statistical significance in mathematical analysis.However,according to the results of the content determination,the gentian is homogeneous.The proportion of oleanolic acid that did not reach the limit of quantification was 15.38%,and the proportion of wild products that did not reach the limit of quantification was 66.67%.Therefore,the content of oleanolic acid in wild products was significantly smaller than that in cultivated products.3.Chloroplast whole genome analysis:The gentian chloroplast genome CP sequence is 149,192 bp in length,with a total GC content of 37.61%,of which SSC(17,269 bp)and LSC(81,353 bp)are composed of 4 typical structures,forming a single IR,The gentian CP genome encodes 125 unique genes,including 84 protein-coding genes,38 t RNA genes,4rRNA genes,and 44 copies of the IR region.Conclusions:1.Appearance1.1 Numerical analysis shows that the wild gentian medicine is whiter than the cultivated gentian medicine,and the proportion of horizontal ring pattern is larger,but there is no difference in other indicators.1.2 Observation with the naked eye shows that the wild gentian medicinal materialsare full in shape,yellowish white in color,and most have main roots,and the fibrous roots are scattered after drying;while the cultivated gentian medicinal materials have shriveled roots,reddish brown in color,and have no main roots.1.3 The gentian content of gentian medicinal materials has a correlation with color.It is speculated that the higher the gentian acid content,the whiter the gentian medicinal materials.2.Chemical composition2.1 The fingerprint similarity evaluation results and HPLC fingerprint analysis showed that there were 16 common peaks in the chemical fingerprints of wild gentian,artificially cultivated gentian,and reference medicinal materials.By reference product designation,5 peaks were identified,which were: raninic acid,argentin,gentiopicrin,argentin,and gentiolide,and most samples were similar by similarity calculation.The degree is greater than 0.9,which indicates that the similarity between wild products and cultivated products is high.2.2 Cyclic ether terpenoids: The content of wild-type raninic acid is significantly greater than that of cultivated products,while the content of wild-type gentisate is less than that of cultivated products.There were no differences in the contents of gentiopicroside,argentin,and argentin.2.3 Polysaccharide content: There is no difference in polysaccharide content between wild and cultivated products.2.4 Flavonoid content: Wild products have less flavonoids than cultivated products.2.5 Triterpenoids: The oleanolic acid content of wild gentian is significantly lower than that of cultivated products.3.Chloroplast whole genome sequencing resultsThe gentian chloroplast genome CP sequence is 149,192 bp in length,with a total GC content of 37.61%,of which SSC(17,269 bp)and LSC(81,353 bp)are composed of 4 typical structures,forming a single IR(25,285bp).The gentian CP genome encodes 125 unique genes,including 84 protein-coding genes,38 t RNA genes,4rRNA genes,and 44 copies of the IR region.One gene with two introns: TRNF-AAA,trn Y-ATA,trn DATC,PSBA,atp F,Trni-TAT,trn N-ATT,rpo C 1,rpl2,ycf 2,ycf 15,ndh B,NDHA,ycf 1,rps12. |
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