| Objective:Through the investigation and quality evaluation of Ek,it was found that the content of active components in Ek could not meet the requirement of pharmacopoeia,which seriously affected the clinical efficacy and market price of Ek.Therefore,on the basis of this study,a systematic study on the quality regionalization of Ek was carried out in order to find the environmental factors affecting Ek and provide a theoretical basis for the artificial tending and fine planting of Ek.On Ek leaf collection(S1-S4)in a different time,different hormone treatment(Me JA,SA),explore the epimedium biomass with Ek,the influence of the external phenotype,internal quality,and materials for the transcriptome sequencing,comprehensive analysis of screening of flavonoids biosynthesis genes,Ek for the study of epimedium contain and physiological and biochemical processes,flavonoids synthesis mechanism,the new variety breeding to provide the reference.Methods:Based on the contents of four flavonoids in Ek,the resource distribution and quality regionalization of Ek were studied by using GIS technology,maximum entropy model(Max Ent)and geographical probe statistical analysis method.Annual north Korea epimedium as material,through the undergrowth bionic cultivation,laid on different time collection(S1-S4)and hormone treatment(Me JA,SA),and USES the ultraviolet spectrophotometer,HPLC method for determination of total flavonoids and four kinds of content of flavone glycosides,portable instrument measuring leaf area,leaf area,high quality computer chromatic meter measure its chromaticity,scales measuring biomass.The samples of Epimedium japonicum leaf treated above were used for transcriptome sequencing using Illumina Hi Seq TM 4000 sequencing platform.The transcriptome splicing,gene annotation,gene expression level and expression difference analysis were used in combination with content determination to screen the regulatory genes related to total flavonoid biosynthesis.Results:1.Ek mainly distributed in the southeastern jilin province and liaoning province northeast,the main chemical component content by the coefficient of variation of annual precipitation,precipitation,annual average temperature,clay quantity is greatly influenced by environmental factors,such as including the herb to determine A herb,and the higher C content distribution in southeast area in jilin province and the northeastern liaoning province,the herb for higher B content distribution area in eastern liaoning province,epimedium glycoside content is higher most region in northeastern liaoning province,A small amount of distributed in the southeast of jilin province.2.The leaf area and biomass,With light source D65,the composition of Epimedium in Korea was mainly correlated with L and A *,and the contents of Epimedium A,B,C and Epimedium were negatively correlated with L value.Is positively correlated with a*value.Under light source A,the contents of icariin A,B,C and icariin were negatively correlated with the values of L and A *(*p<0.05);No correlation with B * value.3.After treatment with different hormones,compared with blank group,there was no significant difference in leaf area and biomass of E.japonicum treated with Me JA,but leaf area,fresh weight and dry weight of E.japonicum treated with SA increased by 24.67%,23.50% and 81.50%,respectively.The content of total flavonoids was increased by 12.89%after treatment with Me JA and 8.44% after treatment with SA.There was no obvious uniform change of the four flavonoid glycosides.It is worth noting that the content of icariin was significantly increased by 67.28% and 113.16% under treatment with two hormones,respectively.4.A total of 18 different c DNA libraries of Ek were constructed.Through transcriptome sequencing,high-quality filtered data of 4980756594-12762476287 bp were obtained,and a total of 366522 Unigenes were functional annotated.Conclusion: The results showed that the contents of flavonoids in Ek varied greatly in different regions,and Ek was mainly distributed in the southeastern part of Jilin Province and the northeastern part of Liaoning Province.With the growth and development of Ek,leaf area and biomass increased,and the content of total flavonoids first increased to the threshold value and then decreased.There was a significant correlation between the chromaticity(quantity)of Epimedium and the intrinsic flavonoids(quality).The 18 c DNA libraries obtained by transcriptome sequencing in this study were of high quality,and the 25 flavonoid biosynthesis-related genes screened were accurate.The above research results provided references for the introduction and cultivation of Ek,artificial cultivation,study on the mechanism of total flavonoid biosynthesis,and new variety breeding. |
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