| Purpose:Gentiana scabra is a famous authentic medicinal material produced in Liao Dynasty.It is called one of the "Six Treasures of Liao Medicine".It has many pharmacological effects,such as clearing heat and dampness,and clearing liver fire.The artificial planting of G.scabra is also the main way to reform the agricultural supply side of Liaoning Province,adjust the agricultural planting structure,promote the development of the health industry and increase the income of farmers.In the long-term practice of Chinese medicine,authenticity has always been a unique comprehensive standard for evaluating the quality of Chinese medicine.Ecological environmental conditions affect the distribution,growth,yield and quality of authentic medicinal materials.Proper habitat conditions are conducive to the formation and accumulation of secondary metabolites in traditional Chinese medicine,thereby affecting the quality of medicinal materials.Environmental factors mainly include topographical factors,climate factors,soil factors and biological factors of the place of production.It is important to understand the formation of authentic medicinal materials and the mechanisms that drive the productivity of medicinal plants in order to optimize the service functions of farmland and natural ecosystems and increase the content of secondary metabolites of medicinal plants.This study takes gentian as the research object,and through understanding the correlation between the comprehensive quality of gentian and ecological factors,it hopes to explore the formation and mechanism of the gentian tract from an ecological perspective,and provide a theory for promoting the large-scale and standardized production of gentian in accordance with.Methods:1.Materials: The experimental materials for this subject were collected in the main distribution areas of G.scabra in Liaoning,Jilin and Heilongjiang.A total of 31 batches of wild or cultivated gentian from different origins were collected(Heihe City,Heilongjiang,Jiamusi City,Heilongjiang,Jilin City,Jilin Province,Jiaohe City,Jilin Province,Qingyuan County,Fushun City,Liaoning Province,Benxi City,Liaoning Province,etc.),the medicinal parts are the roots and rhizomes,and the rhizosphere soil of G.scabra in various producing areas is collected.All samples were identified as G.scabra by Professor Yin Haibo,Department of Chinese Medicine Resources,Liaoning University of Traditional Chinese Medicine.2.Method:2.1 The main distribution areas of G.scabra are Liaoning Province,Jilin Province and Heilongjiang Province,three provinces,11 cities,18 counties and districts to investigate the germplasm resources.Observe,measure and record the ecological environment and plant characteristics in each distribution area,and collect plant samples to collect the rhizosphere soil of gentian from different producing areas.2.2 The moisture content is determined in accordance with the general rules of Part 4 of the Pharmacopoeia of the People’s Republic of China(Practitioner of the People’s Republic of China)(0832 Method 2).Determine the content of ash and acid-insoluble components in accordance with the General Principles of Part 4 of the Pharmacopoeia of the People’s Republic of China 2020(2302 total ash determination method).2.3 Determination of 4 iridoid glycosides in 31 wild and cultivated G.scabra herbal materials from 31 origins by high performance liquid chromatography,namely loganine acid,seriopein,gentiopicroside and seriogenin Comprehensive evaluation of the quality of G.scabra medicinal materials.2.4 High performance liquid chromatography was used to establish fingerprints of wild and cultivated gentian grass from 31 producing areas.2.5 The soil indicators such as mineral elements,PH value,water content,mechanical composition,aggregate structure,organic matter and humus in the soil were determined respectively,and soil factors affecting the quality of G.scabra were screened.2.6 Use Arc GIS software to obtain the ecological factor data of 31 producing areas of G.scabra,including data related to precipitation,sunshine,climate,soil,topography and vegetation type,and screen the meteorological factors that affect the quality of G.scabra;determine the growth environment of gentian Terrain and biological factors,to screen the terrain or biological factors that affect the quality of G.scabra.2.7 Construct a mathematical model of niche suitability of G.scabra to determine the comprehensive influence of habitat factors on quality factors of Ephedra.Results:1.The field survey recorded a total of 322 species,including 38 trees,68 shrubs,and 195 herbs.Among the companion plants of G.scabra,the trees are mostly Pinus tabulaeformis,Acer liaotungensis,Quercus liaotungensis,Shanlihong,Glandular Willow,etc.The shrubs are mostly Hazel,Mountain Rose,Euonymus,Lespedeza,Beef Tripe,etc.There are many herbaceous plants.It is gerbera,Diyu,dragon sprout,berry-leaf potentilla,thawson grass,and so on.The cultivation bases of G.scabra are mainly distributed in eastern Liaoning.Among them,Qingyuan County,Fushun City,Liaoning Province is the largest G.scabra cultivation base in China,and Xinbin County,Fushun City,Huanren County,Benxi City,and Kuandian County,Dandong City,etc.There is cultivation.2.There are significant differences in plant height,number of inflorescences,root length,leaf size and other traits of G.scabra from different producing areas.Among them,G.scabra planted in Yangmuchuan Town,Kuandian County,Dandong City,Liaoning Province are tall and strong with strong main roots.The wild G.scabra in Ying’emen Town,Qingyuan County,Fushun City,Liaoning Province comes next.Among the traits of G.scabra,the largest average standard deviation is the number of inflorescences(σ=0.08);the largest average coefficient of variation is the number of inflorescences(CV = 140.13%).Gentiana plant height(24.53%),inflorescence length(61.46%),inflorescence width(55.03%),number of stems(56.45%),main stem thickness(29.12%),stem middle leaf length(18.46%),stem The coefficient of variation of 9 traits such as leaf width(19.04%),root length(16.05%),rhizome diameter(24.15%)and so on was relatively small.3.The moisture content of 31 batches of G.scabra is 3.62%～8.61%,the total ash content is3.00%～6.67%,and the acid-insoluble ash content is 0.67%～2.67%.The difference in water content between different production areas was 14.66%,the difference in total ash content was 21.70%,and the difference in acid-insoluble content was 31.09%.Among them,loganylic acid has a relatively high content in LD-2 samples(21.6835 mg/g);Swertia citroside has a relatively high average content in LD-28 samples(5.3793 mg/g);Gentiopicrin is in LD-The average content of 16 samples was relatively high(73.0254 mg/g);the average content of sorgenin was relatively high in the LD-2 samples(3.9069 mg/g);the total iridoid content was relatively high in the LD-16 samples(95.2065 mg/g).4.The establishment of the fingerprint of G.scabra showed that the 25 common peaks of 31 batches of test products all appeared within 60 minutes,and by comparing the relative retention time,the 7th peak was designated as loganic acid and the 9th peak was Glycoside,peak 11 is gentiopicroside,peak 13 is seriogenin,the similarity between 31 batches of G.scabra medicinal materials and the control profile is 0.985 to 1.000,and the similarity between batches is close.The variability results showed that the common fingerprint peaks of peak 25,peak 13,peak 15,and peak 16 of gentian from different producing areas have high differences.Among them,the common fingerprint peak 25 of gentian has the largest coefficient of variation(CV = 96.9258%).Total fingerprint peak 1(CV =14.3820%),shared fingerprint peak 9(CV =17.9321%),shared fingerprint peak 11(CV =15.9029%),shared fingerprint peak 12(CV = 24.2712%),shared fingerprint peak 22(CV = 23.3675 %)etc.The coefficient of variation is small.In the results of the systematic cluster analysis,31 batches of gentian from different producing areas can be grouped into 2 categories,which cannot be separated by geographic location.The results of principal component analysis showed that the comprehensive scores(0.60,0.59)of gentian medicinal materials produced in Huanren County,Benxi City were higher than those of other producing areas.Principal component analysis results of G.scabra medicinal materials produced in Huanren County,Benxi City have a high comprehensive score(0.60,0.59)and other producing areas.5.The soil texture of the gentian producing area is divided into five categories.Sandy loam accounted for 58%,silty loam accounted for 16%,loam and loamy sandy soil accounted for less than 16%,and the least was sandy clay.Loam soil accounts for 6%.The average content of inorganic elements in gentian soil is iron>magnesium>calcium>manganese>available potassium>alkali-hydrolyzed nitrogen>zinc>available phosphorus>copper.The total content of inorganic elements in soil is between 9203.5 and 86741.40 mg/kg.The soil water content is between 2.20 ～ 14.50%,and the soil is neutral and acidic.There are significant differences in the aggregate content of soil at different levels.The largest difference is the content of soil aggregates with a particle size of 0.05 to 0.01 mm,and the coefficient of variation is as high as 87.05%.The content of humus in the soil is between 5.59 and 60.37 g/kg,and the content of organic matter is between 4.22 and 45.39 g/kg.Among them,the content of humic acid in the soil is the biggest difference among the components of humus,with the highest variability of 62.95%.6.Through the correlation analysis of 21 quality factors of G.scabra and all habitat factors,the habitat factors that are significantly related to one or more quality factors of G.scabra are screened out,including 22 soil factors and 31 meteorological factors,3 terrain factors and 1biological factor.Soil factors include organic matter,humus,humin,humic acid,fulvic acid,total humic acid and fulvic acid;available potassium,available phosphorus,copper,zinc,manganese,calcium,magnesium,iron;water content,p H value,machinery Compose various structures,2～0.25 mm,0.05～0.01 mm,0.01～0.005 mm,<0.001 mm aggregate structure.Terrain factors include longitude,latitude,and altitude.Biological factor: important value.Meteorological factors include the coldest season average temperature,the warmest season average temperature,the monthly average temperature difference between day and night,the annual average temperature range,the warmest month’s highest temperature,the coldest month’s lowest temperature,the standard deviation of seasonal temperature changes,isothermality,and the wettest Seasonal average temperature,driest season average temperature,annual average humidity,annual average sunshine hours,annual average radiation,annual average temperature,driest month precipitation,rainfall variation coefficient,driest season rainfall,seasonal rainfall,1 ～June rainfall,October～December rainfall,January～December average temperature.7.Niche suitability calculation results show that there are 13 most suitable producing areas with Fi > 0.9,concentrated in Jilin Jiaohe,Tonghua,Fushun Qingyuan and Xinbin in Liaoning;the other 18 producing areas with Fi > 0.8 are more suitable Production area,unsuitable production area is zero.Conclusions:1.The wild resources of G.scabra are scarce,and most of them grow in meadows with sufficient sunlight and little human intervention.The cultivation base of G.scabra is expanding day by day,mainly distributed in Liaodong area,Qingyuan County,Fushun City,Liaoning Province,Xinbin County,Fushun City,Liaoning Province,Huanren County,Benxi City,Kuandian County,Dandong City and other places.Among them,Qingyuan County,Liaoning Province,has the largest gentian planting base with the largest number of households,high yield per mu,and its cultivation and management methods are also the most systematic.2.There are significant differences in plant height,number of inflorescences,root length,leaf size and other traits of gentian from different producing areas.The G.scabra grown in Yangmuchuan Township,Kuandian County are tall and robust,with the best growth conditions,followed by wild gentian plants in Ying’emen Township,Qingyuan County.When choosing the provenance of gentian,you can focus on the number of inflorescences on the gentian plant.The plant height,inflorescence length,inflorescence width,stem number,main stem thickness,stem leaf length,stem leaf width,root length,rhizome diameter and other indicators can also be used to assist in judging the growth of plants.3.The active ingredients of the 31 samples of medicinal materials all reached the standards stipulated in the 2020 edition of the Chinese Pharmacopoeia,and there were certain differences in the content of 4 iridoids in gentian from different regions.Among them,loganylic acid has a relatively high content in LD-2 samples(21.6835 mg/g);Swertia citroside has a relatively high average content in LD-28 samples(5.3793 mg/g);Gentiopicrin is in LD-The average content of 16 samples was relatively high(73.0254 mg/g);the average content of sorgenin was relatively high in the LD-2 samples(3.9069 mg/g);the total iridoid content was relatively high in the LD-16 samples(95.2065 mg/g).The results of principal component analysis and iridoid content determination are consistent,indicating that principal component analysis can more intuitively evaluate the quality of G.scabra.4.Through the establishment of high performance liquid phase fingerprints of G.scabra,a total of 25 common fingerprint peaks were calibrated,and loganic acid,gentiopicroside,seriogenin,and sageripicroside were identified among 4 gentian materials.The similarity between the main active ingredients,31 batches of gentian medicinal materials and the control profile is 0.985 to 1.000,and the similarity between batches is close.The 31 batches of gentian medicinal materials are basically the same in terms of chemical composition,with only minor differences in content.This is also consistent with the reason why cultivated G.scabra materials occupy the medicinal market.5.Correlation analysis results show that there are 57 habitat factors that are significantly related to the comprehensive quality of G.scabra,including 22 soil factors,3 topographic factors,1 biological factor,and 31 meteorological factors.Among them,the content of swervain,medicinal material moisture,ash,acid-insoluble ash,and plant traits are strongly correlated with soil factors.Swerogen is strongly correlated with climatic factors.The accumulation of loganic acid content is affected by the soil factors and climate of the place of production.Factors are affected together.The accumulation of gentiopicrin content is affected by the soil factors,topographic factors and climate factors of the place of production.The fingerprint map is significantly related to the four factors of soil,climate,topography and biological factors.On the whole,the quality of G.scabra is mainly affected by the climatic factors of its production area,followed by soil factors,topographic factors,and biological factors the least.The results of this study can provide a scientific basis for field management during field production of G.scabra.6.Through the establishment of the niche suitability model of G.scabra,it can be seen that the 31 producing areas investigated are suitable for the growth and development of gentian and the accumulation of effective ingredients,of which 13 are the most suitable producing areas,and the main producing areas are concentrated in Jilin Jiaohe and Tonghua,Fushun Qingyuan,Xinbin,Benxi Nanfen District,Xifeng County of Tieling City,Xiuyan County of Anshan City,and Kuandian County of Dandong City in Liaoning.The other producing areas are all suitable producing areas for G.scabra,and all are unsuitable areas.In this study,Liaoning,Heilongjiang,and Jilin were selected as the research objects in the authentic production areas of G.scabra.The results show that the climate and soil are suitable for its growth and development,which can explain its authentic principles. |
