| Eucommia ulmoides Olive., a deciduous tree species classified into Eucommia Oliver ofEucommiaceae, is the relict ancient tree species left after the invasion of Quaternary glacier. As"living fossil plant", E. ulmoide is a type of second-class national key protected species, whoseclosely related species have been extinct and the existing E. ulmoide resources are only savedin China. In this study the barks and leaves are used as experimental materials to analyze thecontent of the pinoresinol diglucoside and chlorogenic acid of E. ulmoide. Additionally, theISSR (inter-simple sequence repeat) technology is applied to explore the genetic diversity ofsamples collected from different origins of E. ulmoide. The main results are as follows：1. HPLC analysis results show that the pinoresinol glucoside in E. ulmoide barks fromYichang, Hubei, Guizhou Zunyi do not meet Pharmacopoeia standards. The samples of Henanand Shaanxi yield the highest level of the pinoresinol glucoside. and the content of plantsamples in Henan and Shaanxi ranks top is more than twice of the standard. Samples of E.ulmoide from Hubei, Guizhou, Chongqing and other places have great homogeneity, while thecontent of pinoresinol glucoside in E. ulmoide barks from Wangcang Sichuan is much lowerthan that from other place in that province. Additionally, the chlorogenic acid contained inleaves of E. ulmoide from Henan and Shaanixi is the highest, while the content in leaves of E.ulmoide within the whole Sichuan is intermediate and stable, and that in leaves from Hubei andGuizhou is the minimum. Therefore, it is important to identify the origions of the samples.2. Eight pairs of ISSR primers were used to amplify E. ulmoide DNAs from15differentorigins, and102DNA fragments were amplified with polymorphic rate at88.2%. The E.ulmoide plants genes within species had a large diversity. The effective number of alleles hit1.781on average and the average gene diversity was0.443. The information index of Shannonwas0.758on average. Genetic similarity at65%. The E. ulmoide in15different habitats can bebroadly divided into three categories, and the genetic distances showed similar with theirgeographical distances. 3. Another finding of this study is the identification of an acidic heteropolysaccharide,EOP-1extracted from E. ulmoide leaves by partial acid hydrolysis, methylation analysis et al.The molecular weight is about6×105, and consists of galacturonic acid, galactose, rhamnose,mannose, glucose and arabinose. In this polysaccharide, the content of glucose, galactose wasrelatively high, which included main part of homogalacturonan fragments and residues of sugarconstituents were mainly D-GalpA, the backbone was composed of1,4-linked-D-GalpA and1,3-D-Galp、1,4-D-Galp、1,5-L-Araf were the mainly consititution of the branch. Thebioactivity of this acidic heteropolysaccharide deserves a further anlaysis.4. Based on the pharmacological components of E. ulmoide, polysaccharide was one ofimportant effective components. The compositions of cell wall of E. ulmoide from differentorgans and different areas of production are compared in this paper. Four components of lignin,cellulose, pectin and hemicellulose of cell wall of E. ulmoide in different organs (leaf, root,bark) in five different areas from the North to the South China, and the neutral monosaccharidein pectin and hemicellulose was further analyzed. The results showed that the total content ofneutral sugar was the highest in leaf tissue and showed gradual decline in root and bark; thecontent of uronic was is also the highest in leaf tissue, followed by the root, that it was thelowest in the bark. The results showed the fraction of cellulose indicated gradual decline in root,bark and leaf. The content of lignin showed an increasing trend in leaf, root and bark. totalneutral sugar and uronic acid content of the leaf was higher than that of the bark and root whichindicated that the bark could be replaced by the leaf.The varied contents and ratios of pharmacological components including neutralmonosaccharides in the leaves and barks of E. ulmoide from different organs and from samplesin different origins demonstrate the needs to develop cut-edge technology to fully use E.ulmoide resources. The similarity of the contents of pinoresinol glucoside and chlorogenic acidof the E. ulmoide samples with their genetic distances provides the opportunity to developmoleculat markers to identify the origins thus to control the quanlity of medicines. The geneticvaribility and enviromental factors influence the comttent of the pharmacological components thus it is required to disseminate the effects of genetic and enviromental determination on thefinal output of the medical herb. |
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