| Tetrastigma hemsleyanum Diels et Gilg(Vitaceae),belongs to the family Vitaceae.It is a folk and endangered medicinal herb in southern China,The entire plant,especially its root,has some medical efficacies,such as antipyretic,detoxification for children.HPLC fingerprint combined with multi-index composition quantitative analysis and chemometrics methods were used in this study to evaluate the comprehensive quality of different origins,different root parts and different storage conditions of T.hemsleyanum.Otherwise,its rapid propagation system using tissue culture technology was established.This study afforded more comprehensive references for T.hemsleyanum quality control and seed-breeding so as to expend the sustainable utilization and resources protection of T.hemsleyanum.1.We determined the water content and extractum content of T.hemsleyanum from Hangzhou,Jinhua,Zhoushan,Guangxi and Guizhou,respectively.The result showed that water content varied from 6.4% to 11.2%,and extractum content varied from 6.7% to 14.9%.Then we measured the water content and extractum content of T.hemsleyanum root powder from Hangzhou Tianhui,Shou Xiangu and Huadong Traditional Chinese Medicine CO.,Ltd.The water content of all powders were under 11.2%,and their extractum content were higher than 6.4%.2.Using HPLC to compare the fingerprint difference of T.hemsleyanum from six origins,including Hangzhou,Taizhou,Jiangxi,Guangxi and Chongqing,the control fingerprint was generated in this study.More importantly,it was found that Hangzhou,Taizhou,Jiangxi,Guizhou and Chongqing were classified into one cluster while Guangxi and Guizhou were other by chemometrics methods including similarity analysis,cluster analysis and principal component analysis(PCA).Moreover,PCA of 25 characteristic peaks showed that peaks 19,14,15 were the main contributors to principal component 1(PC1),peaks 1,7,4,18 were the main contributors to PC2,and peaks 6 and 21 were the main contributors to PC3.These results demonstrated that the chemical constitutions which these peaks reprencent were the main characteristic components of T.hemsleyanum,but it still needs further study.3.Polydatin,piceatannol,nicotiflorin,resveratrol and kaempferol was selected as index components.Based on the analysis of HPLC results of T.hemsleyanum from six origins,it was found that three phenolic acids content(polydatin,piceatannol,resveratrol)were higher in samples from Chongqing and Hangzhou than Guangxi and Guizhou while the content of two flavonoids(nicotiflorin,kaempferol)were higher in samples from Guangxi and Guizhou than Hangzhou and Thaizhou.These results were consistent with cluster analysis in fingerprint study.Then the content of five index components in four parts of T.hemsleyanum root tuber was determined.Results implied that the reducing order of three phenolic acids content was from taproot to fibrous root to taproot without velamen to velamen.And reducing order of two flavonoids content was from fibrous root to taproot to taproot without velamen to velamen.It is noting that the content of total five index components in fibrous root could reach as much as 84.69% of taproot.It implied that the fibrous root was worth developing as medicinal part of T.hemsleyanum.In addition,we compared the contents variation of effective components in T.hemsleyanum root stored at 25℃ and 4℃ with time.The results showed that content of polydatin and piceatannol were declined dramatically after 1 month when stored at 25℃ while content of quercetin and kaempferol were declined not much.Samples stored at 25℃ became putrid after 2 months while samples stored at 4℃ became putrid after 5 months,and contents variation of effective components were not significant.Therefore,we suggest that T.hemsleyanum fresh root should be stored at 4℃ and used within 5 months after vacuum packing.4.We established the micropropagation system of T.hemsleyanum using stem segments with axillary buds.The results showed that the semi-lignified stem segments are better than stem apex as explants,the suitable culture mediums for bud induction,bud proliferation,and rooting were MS+1.5 mg/L 6-BA+0.025 mg/L NAA,MS+1 mg/L 6-BA+0.025 mg/L NAA+0.025 mg/L GA,and 1/2 MS+1.5 mg/L IBA,separately.In addition,the survival rate of transplants was more than 90%.With lower hormone concentrations,user-friendly control,and high efficient breeding,the micropropagation system is totally suitable for large-scale and standardized production of seedlings of T.hemsleyanum. |
