| Dendrobium officinale is a traditional Chinese Medicine which was first recorded in Shen Nong's Herbal Class has been used as folk medicine for thousands of years.In the wild,this plant is near extinction.The Dendrobium officinale sold in the medicinal market was cultivated in the greenhouse.At present,there existed many issues of this cultivated materials such as provenance,hybridization with different Dendrobium genus,which dramatically restrict the quality of this medicinal plant.The quality,external properties and genetic information from different cultivated population was different.The characterization and identification of external properties of the plants are two basic and direct way to understand different germplasm resource,and they are also important index to evaluate the quality of the plant resource.The SSR fingerprint analysis is one of the most accepted methods for species identification.Through establishing the chromatography fingerprint and SSR fingerprint,the genus,species and individual of the Dendrobium officinale can be identified.At present,the pesticide effect of the Dendrobium officniale is based on the content of the polysaccharide.However,the polysaccharide content in this plant diverse dramatically from different plantation environment and location.It is critical to study this diversity of this plant.In this study,we collected 392 samples of the Dendrobium officinale from Guangxi,Guangdong,Zhejiang,Anhui,Fujian,Yunnan,Guizhou and Sichuan provinces.In order to rearrange,determine the key germplasm resources and provide certain references for application,the agronomic character,polysaccharides content,SSR fingerprint and DNA ID of these samples was described,tested and established.Objective:Through collecting the Dendrobium officianle samples from different places of production,arranging the agronomic character,establishing the SSR fingerprints and DNA IDs,determining the polysaccharide content differences,the intrinsic relation between these index and places of production was analyzed.Method:1.To analyze the agronomic character of the Dendrobium officinale through principle component analysis method,and the main character components were screened out.And using cluster analysis to build clustering tree of the different population of this plant.2.The DNA was extracted by CTAB reagent.The DNA was amplified by PCR,and the polymorphism of the PCR product was analyzed through analyzing the band differences by 12%silver staining polyacrylamide gel electrophoresis.3.The polysaccharide content was detected by PMP-HPLC method.The content of mannose,glucose and total sugar was used as variable to analysis the relation of the samples from different populations.And the clustering tree was constructed.All calculation was done by using SPSS 23.0 software.Results:1.The coefficient variation of the stem height and leaf area was higher,and most of the external properties were significantly related.Each of the variables was the key external to distinguish from different populations.The line L3 divided these 38 populations into 6 classes.This result was determined by the places of production.2.4 pair of the SSR primers was applied for amplification.19 polymorphic bands were spotted and each primer can detected 4.75 bands in average.UPGMA mathematical method was used to analysis the cluster of the 29 populations of the plant Dendrobium officinale from 9 places of production.These 29 populations can be divided into 4 classes.Samples from Yunnan,Guizhou and Sichuan provinces were grouped together.Samples from Anhui province grouped individually,while samples from Guangxi province also grouped as one class.Samples from Guangdong province Danxia area,Zhejiang province Yongkang district,Leqing district and Taining district were grouped as one class.3.The polysaccharide content and the mannose and glucose ratio of the Dendrobium officinale from different populations was different.The conjugation line L3 divided these 29 populations into 4 classed,The samples named YK1 to YK3,YN3,YN-6 and GPH were grouped as A group.Samples named as ZJ,A6,A1,A5 and TN2,GZ were grouped as B group.Samples named as A2,A4,TN1 and SM were grouped as C group.Samples named as DX1 to DX4,YN2,YN4 to YN5 and GPL was grouped as D group.Conclusion:1.Through analyzing of the economical character of the Dendrobium officinale,it was found that this plant was extremely diverse in genetically.Through cluster analysis,we found that the classification of the populations was significantly related to the places of production,stating that the populations cluster results exhibit remarkable relation with the places of production.This character can be applied as a way to identify the germplasm resource of Dendrobium officinale.2.The ID built by SSR fingerprint with these 4 primers was closely related to the places of production.Through establishing the DNA ID and genetic similarity of the Dendrobium officinale from different populations,the germplasm information of Dendrobium officinale can be defined.3.Through analyzing the polysaccharides content of the 29 populations from 9 places of productions,the polysaccharides content as well as the mannose glucose ratio of this plant was significantly different when comparing samples from different places of production.The samples with better economical character had higher polysaccharides content.Those better populations can be promoted as cultivation germplasm. |
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