Genetic Diversity And HPLC Fingerprint Analysis Of Dendrobium Officinale Kimura Et Migo

In this study, genetic diversity and genetic relationship of twenty Dendrobiumofficinale Kimura et Migo from different places were reseached by ISSR molecularmarker, in order to lay a foundation for genetic background and variety breeding. Themedicinal components in stem and leaves of Dendrobium officinale Kimura et Migofrom10different producing areas were detected by HPLC-UV,and the HPLCfingerprinting in different parts of Dendrobium officinale Kimura et Migo werestudied. To understand the effect of producing area and harvest time on the results ofquantitative analyses, the variation of medicinal components in Dendrobium officinaleKimura et Migo from ten different regions and different parts and harvest time wereanalyzed. The primary research results are as follows:1The investigation of the genetic diversity by ISSR molecular marker. Twelvepolymorphism primers that selected from100ISSR primers had amplification167DNA bands in total in which154were polymorphism bands, and the ratio ofpolymorphism in average was92.21percent. By POPGENE32analysis, the averageof Observed number of alleles(Na) is1.9701, effective number of alleles(Ne) is0.5373, Nei’s genetic diversity(H) is0.3646and the Shannon’s information index(I) is1.6408. The genetic similarity(GS) for twenty varieties experimental material by DPSranged from0.41to0.82. In the process of Cluster Analysis,18copies of Dendrobiumofficinale Kimura et Migo were gather into a large class, but some materials that havethe close geographical position were not necessarily gotten together in further study.The results illustrated that the test materials were more complex, the geneticbackground is rich.2The HPLC fingerprint method for the medicinal components in Dendrobiumofficinale Kimura et Migo was established. Chromatographic conditions: WatersC18(5μm,4.6×250mm), the mobile phase: acetonitrile-0.05%phosphoric acid(gradientelution), flow rate:0.8mL/min, the detection wavelength:225nm, the injection volume:20μL, column temperature was25℃, analysis time:75min.3Obtained the HPLC fingerprinting in stem and leaf of Dendrobium officinaleKimura et Migo. Fourteen common peaks (the9th peak was naringenin) in thefingerprints of stem were determined. Thirteen common peaks (the8th peak wasnaringenin) in the fingerprints of leaf were determined. The results showed thesimilarity between the HPLC fingerprints in the same part (stem, leaf) from differentDendrobium officinale Kimura et Migo, the difference between stem and leaf from thesame sample was significant.4There were significant differences of the main medicinal ingredients inDendrobium officinale Kimura et Migo from different areas. The contents ofpolysaccharide in stem that from Guangxi-Nanning, Zhejiang-Xiaoshan,Zhejiang-Hangzhou, the contents of total flavonoids in stem that fromZhejiang-Xiaoshan, Guangxi-Nanning, the contents of alkaloid in stem that fromZhejiang-Leqing, Zhejiang-Ruian were higher than other areas. The contents ofpolysaccharide in leaf that from Zhejiang-Ruian, Zhejiang, Zhejiang-Leqing, thecontents of total flavonoids in leaf that from Guangdong-Raoping, Fujian-Longyan,the contents of alkaloid in leaf that from Guangdong-Raoping, Fujian-Longyan,Zhejiang-Ruian were higher than other areas.5There were significant differences of the main medicinal ingredients inDendrobium officinale Kimura et Migo in different harvest time. Content ofPolysaccharide and total flavonoids were higher in March, but content of alkaloid washigher in October.