| Objective: To investigate the germplasm resources of Salvia miltiorrhiza Bunge in Anhui province,and to explore the morphological changes of Salvia miltiorrhiza in different populations,and to analying the foundation of the selection of fine germplasm.To determine the content of water-soluble active ingredients and fat-soluble active ingredients in 14 areas of Salvia miltiorrhiza,the correlation between the content of active ingredients and traits and growth environment was analyzed.Using chemical fingerprints,with its main origin of Salvia miltiorrhiza as a control,the similarity evaluation system of 18 different origin salvia miltiorrhiza was established and the system clustering analysis was carried out.The results provide a scientific basis for the differential identification of Salvia miltiorrhiza germplasm and the quality evaluation of medicinal materials.Methods: Combined with the fourth national survey on Chinese meteria medica resources,mainly to Wandong,Wannan and Wanxi mountain wild Salvia miltiorrhiza resources collected on-site investigation;and then unified planted in the school garden of Salvia miltiorrhiza morphology and micro-morphological characteristics of the statistical the total DNA of Salvia miltiorrhiza genome was extracted by modified CTAB method.The gene sequence and chloroplast gene sequence were analyzed by PCR amplification technique.The genetic structure of Salvia miltiorrhiza was analyzed.The HPLC reaction conditions were as follows: acetonitrile-0.02% phosphoric acid as the mobile phase gradient elution,detection wavelength of 270 nm,flow rate of 1.0m L·min-1,column temperature of 22 ?,determination of Danshen fat-soluble cryptotanshinone,tanshinone ? and tanshinone ?A,and the correlation between the content of liposoluble and traits was compared.The HPLC reaction conditions were as follows: acetonitrile-0.1% phosphoric acid(22:78),the detection wavelength was 286 nm and the flow rate was 1.2 m L·min-1,the column temperature was 22 ?,and the content of water-soluble salvianolic acid B was determined.The gradient was eluted with acetonitrile-0.1% phosphoric acid as the mobile phase.The detection wavelength was 270 nm and the flow rate was 1.2 m L·min-1,and the column temperature was 30 ?,fingerprints of Salvia miltiorrhiza were established,and the data clustering analysis was carried out by "Chromatographic Fingerprint Similarity Evaluation System"(2012 A Edition)and SPSS 19.0 software.Results: There were significant differences in the morphological differences between the different populations,and the differentiation trend was observed with the latitude and longitude.The morphological differences of plant height,leaf length,leaf length,leaflet width and umbrella inflorescence were observed in morphological aspects.In addition,there were some changes in corolla,leaflets and there was a certain difference in the color of the root bark.micro-morphological observation showed that the leaf color and small leaf traits had obvious difference.The contents of three kinds of tanshinones in salvia miltiorrhiza in 14 regions were different from that of salvianolic acid B,and the content of liposoluble in Salvia miltiorrhiza was closely related to its traits,and the content difference was related to the environmental changes in different regions.The HPLC fingerprints of Salvia miltiorrhiza in different habitats were established,and nine of them were identified.The similarity was only eight,and the difference was more than 0.90.There was a big difference between Salvia miltiorrhiza of different habitat.Conclusion: In this paper,by comparing the morphological differences and their genetic diversity of Salvia miltiorrhiza in different habitats in Anhui Province,it was found that the genetic relationship among Salvia miltiorrhiza had changed,and there was a certain genetic distance,and the different environment had a great influence on the germplasm.The comparison of the contents of the active ingredients of Salvia miltiorrhiza Bunge and the correlation study showed that they had a positive correlation with the origin cluster and the correlation.The fingerprint identification method established in this experiment is simple and reproducible,which can provide a strong basis for the quality control of Salvia miltiorrhiza. |
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