Synthetical Evaluation For The Germplasm Resources Of Ophiopogon Japonicus In Sichuan Basin

The medicinal materials of O. japonicus is the dry root of Ophiopogon japonicus(Thunb.)Ker-Gawl. in Chinese Pharmacopoeia (2005 Edition), which is mainly distributed in Sichuan, Zhejiang and other places of China. In this study, we have collected wild resources of O. japonicus which were came from 26 different areas in hilly area and basin mountain of Sichuan, western Sichuan plain, and the Chongqing region. We have established the chromatographic fingerprints on wild germplasm resource of 0. japonicus and developed the study of chemical pattern recognition. At the same time, we created multi-level quality evaluation system for O. japonicus integrate with morphological parameter, content of major categories compositions and amino acid, fingerprints and other multi-dimensional information, to conducted a comprehensive evaluation analysis on wild resources of O. japonicus in different parts of Sichuan Basin. Now we have preliminarily screening out a number of high-quality species resources of 0. japonicus and provided a valid reference for the works of stoichiometric classification, scientific quality evaluation, effective quality control and breeding of new varietyThe specific results of these studies are as follow:1. Morphological characteristics of germplasm resources of 0. japonicus in Sichuan basin.The results showed that there are extensive variations in morphology, the most significant variance lay in the fresh root weight, dry root weight, and fresh leaf weight, dry leaf weigh, the least significant variance lay in the root length and leaf length; Meanwhile, morphological characteristics of O. japonicus have significant relationships; The results of principal component analysis and cluster analysis showed that the first principal component reflected the characteristics of plant quantity, the second principal component reflected the characteristics of stem leaf, and the third principal component reflected the characteristics of earthnut, all of these were main factors which resulted in morphological variation of O. japonicus. cluster analysis based on the morphological characters showed that the 26 germplasm resources of O. japonicus in Sichuan basin were divided into 4 types. The differences of principal component among the types of O. japonicus were obvious. It is able to select the specific samples from the wide resources of wild 0. japonicus according to the different needs of development and utilization.2. Content of main components in the germplasm resources of O. japonicus in Sichuan basin.To determine the content of main components(such as saponin, flavonoid and soluble polysaccharide) in different germplasm resources of O. japonicus in Sichuan basin with spectrophotometric determination. The results showed that, to the samples of O. japonicus from different areas, their average content of saponin is 2.55%, the content in the sample which was came from Ziyang Jianyang is the highest, it is reached to 6.17%; The content in the sample which was came from Guang'an Wusheng is the lowest, it is merely 6.17%; The relative standard deviation between the samples is 60.47%, the difference is enormous. The average content of flavonoid is 0.12%, the content in the sample which was came from Leshan Shizhong is the highest, it is reached to 0.23%; The content in the sample which was came from Luzhou Shizhong is the lowest, it is merely 0.04%; The relative standard deviation between the samples is 60.47%, the difference is enormous. The average content of soluble polysaccharide is 16.68%, the content in the sample which was came from Luzhou Shizhong a is the highest, it is reached to 40.33%; The content in the sample which was came from Guang'an Shizhong is the lowest, it is merely 0.04%; The relative standard deviation between the samples is 57.34%, the difference is enormous. There are large differences for the contents of major categories compositions among the samples of O. japonicus from different areas, the quality variation for the wild resources of O. japonicus is evident.3. Contents of amino acids in the germplasm resources of O. japonicus in Sichuan basin.The contents of amino acids were determined by HITICHI L-8800 automatic analysis instrument. The results showed that, there are so many kinds of amino acids in the O. japonicus, the sort of amino acids among such resources were different, their contents of amino acids and essential amino acids were higher than common vegetables and Fruits. The contents of essential amino acids in most samples were lower than the FAO/WHO recommended pattern. The contents of amino acids with special effects and sensitive flavor effects, such as Pro, Glu, Phe, Met, were higher than other amino acids. It is unfavorable to eat directly as commen foodstuff, it is favorable to use as natural raw materials on nutritional supplements, fresh-keeping agents, food additives, flavoring agent; There were plentiful variations among the germplasm resource of O. japonicus in Sichuan Basin, the differences of amino acids distribution among different samples were huge, it can be able to select resources of O. japonicus with kinds of uses, and to be utilized synthetically. 4. Chromatographic fingerprints of the germplasm resources of 0. japonicus in Sichuan basin.To used ethanol as a solvent for ultrasonic extraction and defined the fat-soluble components as main examination, the chromatographic fingerprint was established for evaluating and controlling the quality of germplasm resource of 0. japonicus in Sichuan Basin by reversed-phase high performance liquid chromatographic combined with gas chromatographic mass spectrometry. The results showed that each sample was characterized by the peak area of 28 HPLC peaks and 13 GC peaks in each program and these peaks were employed for hierarchical cluster analysis. Furthermore, the discrimination of the sample from different regions was achieved by hierarchical cluster analysis via recognizing the 16x41 datamatrix. This was the first report of hierarchical cluster analysis of wild germplasm resource of O. japonicus according to their Chemical fingerprint. Thus, the results proved it is a simple, rapid and accurate method suitable for the quality control of the traditional Chinese medicines.5. Synthetical evaluation for the germplasm resources of 0. japonicus in Sichuan basin.According to the Main contents and Yield traits, the wild germplasm resources of 0. japonicus from different areas of Sichuan basin were comprehensively compared by the SPSS 17.0 software with principal component analysis and cluster analysis. The results showed that, The six samples of Ziyang, Jianyang, Leshan, Yibin,Chongqing, Mianyang, their comprehensive evaluation value of quality were higher than others, and the sample of Ziyang have the best quality, the sample of Dazhou have the worst quality, the results of the cluster analysis to raw data were also shown a similar law as principal component analysis. The wild resources of 0. japonicus in Sichuan Basin is rich, there were much differences among their quality; the method, through principal component analysis to study the comprehensive evaluation of the 0. japonicus quality,was reliability and the results of cluster analysis was also support their Conclusions,it can be able to provide a reference to Select high-quality resources of O.japonicu.