Germplasm Resources Evaluation And Genetic Diversity Analysis Of Desmodii Styracifolii

Desmodii Styracifolii comes from dried ground grass of Desmodium styracifolium(Osb.)Merr[1].? Lingnan herbs ? record had named “ Dichondra repens” as Desmodium styracifolium,which were widely used by local people,according to the reported literatures[2],they mainly distributed in south place,such as Guangdong,Guangxi,Yunnan,Hainan province.And in Guangdong,Guangxi Zhuang Autonomous Region,they were often referred to “Herba Lysimachiae”,“horseshoe grass” or “phoney peanuts”.The main clinical effect of Desmodium styracifolium are heat and dampness clearing,diuresis and stone removing,and they were commonly used in the treatment of urinary stones,cholecystitis,gallstones and other diseases [3].By studying on the implantation conditions of Desmodium styracifolium in Guangdong Province and Guangxi Zhuang Autonomous Region,these two province always have large-scale cultivation this kind of herb have already formed a relatively stable cultivars.Wherein,Guangxi province Guilin city Lingui County provides the largest planting scale through artificial cultivation,and the annual output were beyond 2,000 tons,which is currently the largest production area of Desmodium styracifolium 's cultivation.HPLC-UV has been used for the quantitative analysis of five flavonoids compounds(Schaftoside,Vicenin 2,Vicenin 3,isoorientin,isovitexin)in Desmodium styracifolium are using.It was found that the content of Schaftoside were higher than the pharmacopoeia's standard,and the cultivars of Desmodium styracifolium contained five flavonoids were obviously higher than those of wild and semi-wild herbs,and the cultivars' quality were more stable and easy to control.23 samples from different harvest time and different production areas of Desmodium styracifolium were analyzed by HPLC technology to establish HPLC fingerprint.A total of 14 peaks were compared with the chemical standards,and we confirmed six major chemical compositions.Using chromatographic fingerprint similarity evaluation system(2004A)analyzed sample chromatograms,the similarity between HPLC profiles of Desmodium styracifolium(from different harvest time and different production areas)and the control profiles is 0.952-0.992.Principal component analysis results of a total of 23 herbs sample peak areas had shown that the cumulative contribution of the first and second principal components factor was 88%.Among them,6,8,11,12,4,14,3 peaks had greater contributions to principal component factor 1,while the first peak had greater contributions to principal component factor 2.Using SPSS software,hierarchical clustering WARD method,Euclidean distance,chi-square measure and squared Euclidean distance clustering methods analyzed the experimental results,showing that G8,G9,G10,G15,G16 and G23 as a class,the remaining samples as another type,and the results were in accordance with the main component analysis results.In the other categories,G7,G11 and G21 were in the same class;G17,G18,G19 and G20 gathered in a class,they are negatively correlated with principal components 1 and 2,but close to the mid-value of position.G4,G5,G6 those three samples' distances were close and agreed with the main component analysis results.The genetic diversity and phylogenetic relationships of Desmodium styracifolium in 15 producting areas of Guangdong Province and Guangxi Zhuang Autonomous Region were analyzed by using ISSR molecular marker technology.Selecting 7 primers from the ISSR universal primers amplified the genomic DNA from Desmodium styracifolium,51 clear bands were observed under UV light,among them,the number of diversity bands were 41.The amplification results were analyzed by using Popgene 1.32 software,Shannon diversity index(I)was 0.3,Nei's genetic diversity index(H)was 0.2464,coefficient of genetic differentiation among populations(GST)was 0.1238,gene flow(Nm)was 3.5397.The above results indicated that both cultivated and wild species of Dichondra repens Forst in Guangdong and Guangxi Zhuang Autonomous Region had higher genetic diversity.According Nes' s genetic similarity differences,NTsys software was used for UPGMA cluster analysis.The results declare that Desmodium styracifolium samples collected from Lingui country,Guilin city,Guangxi Zhuang Autonomous Region were in the same classification;samples from Liuzhou city,Rongan Siding,Siding town,Sanpo Village and Laibin city,Jilin village were in one group and then with samples of Guilin city Lingui District became the same class;the samples from Zhanjiang City in Guangdong Province were together and then with Guangxi's samples were into one category;two kinds of samples in Guangdong Huizhou region were into one classification,and then with herbal samples from Guangdong Province,Shanwei City,Zijin County,Huazhou city were into a large class.Those analysis results illustrate that Desmodium styracifolium produced from Guangdong and Guangxi provinces have certain genetic differentiation,and reflecting the genetic relationship have a certain relevance with geographic distribution.