Pharmacognosy Identification Research Of Dendrobium Huoshanense

Objective: The purpose of this research is to study the biopharmaceutical characteristics of Dendrobium.Huoshanense medicinal materials and its closely related species which are all collected in Chinese Pharmacopoeia,but also to establish a quality evaluation system for them.We expect our result could provide a basis for its cultivational quality control and sustainable production of D.huoshanense medicinal materials.Methods: The similarities and differences among D.huoshanense medicinal materials and other species in genus Dendrobium collected in Chinese Pharmacopoeia compared by physicochemical and molecular methods such as microscopy observation,thin layer chromatography detection,HPLC fingerprinting anlysis,and SSR molecular marker identification.The differences of medicinal quality come from different growth years and different cultivation bases in D.huoshanense medicinal materials were evaluated by HPLC fingerprint chromatograms,and the data were analyzed by similarity evaluation detection,cluster analysis and principal component analysis.The contents of four chemicals such as rutin,ginsenoside,syringic acid,and naringenin in D.huoshanense medicinal materials were determined by external standard method to identify the different species in genus Dendrobium.Results: There are significant differences not only among the six detected Dendrobium dry drugs in their shapes,sizes,surface colors and their transection colors but also in the color,microscopy character,HPLC finger-print photograpy and SSR molecular marker among these medicinal powder.The similarity of fingerprint chromatogrames among the annual and biennial,three year old cultivated D.huoshanense medicinal materials is less than 0.900,and the similarity among different growth years in imitating wild D.huoshanense medicinal materials are greater than 0.900.The higher cultivate age,the higher the similarity among samples was observed.The similarity of D.huoshanense medicinal materials from different cultivational bases were all less than 0.900,cluster analysis showed that there were differences for D.huoshanense medicinal materials at different growth years as well as different cultivation bases.The contents of four components in three-year-old greenhouse cultivation products of D.huoshanense medicinal materials is highest,while there's no difference between the annual and the biennial years plants.The content of four components in the three-yearold D.huoshanense medicinal materials in the wild products was the highest,but the contents of rutin,ginseng phenol,and naringenin remained basically unchanged,and had no significant relationship with the growth period.Sixteen pairs of primers were screened by PCR amplification.The inter-species amplification results showed that the primer c299343 had a specific band around 300 bp,which could be used to identify D.huoshanense medicinal materials from other Dendrobium species.The other pair of primers was c299343,and the two strains could be amplified from the D.huoshanense No.1,and the D.huoshanense No.2 strain could amplify a band.Conclusion: In this study,we systematically studied the traits,microscopic,thin-layer,high-performance liquid chromatogram and molecular marke identification of D.huoshanense medicinal materials and other closely related species.It was found that D.huoshanense medicinal materials and other plants from genus Dendrobium had significant differences in medicinal properties,thin layers and fingerprints chromatogram.The thin layer chromatography and HPLC fingerprinting method established in this experiment are simple and reproducible.The PCR of SSR sequence can realize accurate identification of interspecies and intraspecific of D.huoshanense.It has the advantages of simple operation,stability and reliability,and a wide range of applications.The above-mentioned identification method is of great significant for the quality control of D.huoshanense medicinal materials and the sustainable development of its industry.At present,the cultivation system of D.huoshanense medicinal materials in greenhouse has not been unified yet.The differences in sources,cultivation substrates,and fertilizers from different cultivation bases can lead certain differences in the clustering analysis of medicinal materials.However,the imitation of wild cultivation was relatively stable due to the effects of undisturbed factors as well as influence from natural environment.The content of 4 chemical constituents in D.huoshanense medicinal materials in the different age was quite different.Among them,the contents of four components in three years were higher than those in annual and biennial,which corresponded to the results of cluster analysis.Our finding confirmed that three-yearold stems of D.huoshanense medicinal materials were traditionally used as medicine is reasonable.