| Glechoma longituba (Nakai) Kupr.(Labiatae) is an herbaceous medicinal plant. Its dried aerial part, Glechomae herba, is a standard medicinal material in the Chinese Pharmacopoeia (2010edition) and is used for the treatment of cholecystitis, gall-stone, kidney stone, etc. So far, there are few reports on the collection, establishing of germplasm bank and quality evaluation of the G. longituba germplasm resource. In present doctoral dissertation, the theory and method of Medicinal Botany, Science of Medicinal Plant Resource, Food Nutriology, Medicinal Plant Ecology as well as modern inorganic element analysis and Molecular Biology method were synthetically applied. Based on the multidiscipline theory and method, we collected different G. longituba germplasm resources from20provinces in China and compared the biological characteristics, nutrient quality and pharmaceutical quality. And we explored the genetic relationship among germplasm resources based on ISSR and AFLP markers as well as ITS sequence analysis. In addition, dynamic change of growth characteristic, contents of nutrient elements, heavy metals, and bioactive components in G. longituba from Zijinshan Mountain was investigated. The main results are as follows.1. Fifty-eight G. longituba populations were collected from main contribution regions in China. The main reproductive mode of wild G. longituba is vegetable propagation, and sex reproduction is possible under appropriate microclimate conditions. There are marked difference in morphological characteristics among G. longituba populations. The correlation analysis showed that leaf diameter was linearly correlated with petiole length and internodes diameter, and internode number was linearly correlated with petiole length. Additionally, internodes length and bud number showed linear correlation.2. Present study was conducted to assess molecular genetic diversity among54G. longituba populations in China by ISSR markers for the first time. Eighteen ISSR primers which were screened from47ISSR primers generated a total of189bands and varied in size from200to500bp. Of the189bands obtained,148bands were polymorphic (78.31%). The results of ISSR marker showed that G. longituba populations from different areas of China exhibited abundant genetic variation.3. Simultaneously, molecular genetic diversity among54G. longituba populations was evaluated based on AFLP markers. Eight primers which were screened from64ISSR primers generated a total of1199bands and varied in size from581to734bp. Of the1199bands obtained,1184bands were polymorphic (99.75%). The results of AFLP marker showed that G. longituba populations from different areas of China exhibited abundant genetic variation.4. In terms of pharmaceutical quality and safety, quality evaluation of41G. longituba populations was carried out based on contents of water, total ash, acid-insoluble ash, ethanol-soluble extractive, total flavonoids, ursolic acid (UA) and oleanolic acid (OA), and contents of five heavy metals, viz:Cu、Pb、Cd、As、 Cr. The results showed that different G. longituba. populations showed significant variation in contents of ethanol-soluble extractive, total flavonoids, UA and OA. In generally, the high-quality G. longituba populations that are rich in ethanol-soluble extractive, total flavonoids, ursolic acid, such as the population from Jianghuai Hilly region and Yunnan-Guizhou Plateau, should be interesting to candidates for future research aiming to elevate major bioactive compounds levels in cultivated G. longituba material. Different G. longituba populations showed significant variation in contents of Cu、Pb、Cd、As、Cr in aerial parts. Lead content ranged from0.025to22.300mg·kg-1. The copper content in different G. longituba populations met Chinese standard with the average value of15.562mg·kg-1. The contents of cadmium and chromium ranged from0.013～1.013mg·kg-1, and0～17.75mg·kg-1, respectively. Arsenic content in G. longituba populations met Chinese standard except for samples from few locations. In generally, G. longituba populations from Southwest China and Huaihe region contained lower heavy metals contents.5. Nutritional quality evaluation of41G. longituba populations was carried out based on contents of soluble protein, soluble sugar and fat as well as contents of ten nutrient elements, viz:P, K, Ca, Mg, Zn, Cu, Fe, Al, Mn, and B. The overall trend was that the contents of soluble protein, soluble sugar and fat in samples from South China was higher than those from North China, while the fat content in ones from East china were higher than those from West China. The variation patterns of Ca, Mg, and K were similar. In the case of microelement contents, Zn content changed following the similar pattern of Cu content. The similar variation tendency was found in Fe content and Al content, Mn content and B conten. 6. The growth of wild G. longituba in Zijinshan Mountain was carried out in continuous three years. G longituba emerged in early February; blossomed after rapid growth in March; seeds matured in April; and crept in May. The aerial part could live safely through the winter. Its growth stages could be divided into six stages, viz:emerging, vegetable growth, florescence, seed maturity, creeping, and wilt.7. The contents of water and ethanol-soluble extractive in G herba from Zijinshan Mountain met Chinese pharmacopeia standard irrespective of harvesting time. And the general dynamic change of total flavonoids, ursolic acid and oleanolic acid showed double-peak curve. The optimum harvesting time should be April-August.8. The dynamic change of contents of soluble protein, soluble sugar and fat as well as contents of8nutrient elements, viz:P, K, Ca, Mg, Zn, Fe, Mn, and B. in wild G. longituba from Zijinshan Mountain was measured. The resulted showed that the contents of soluble protein, soluble sugar and fat in G longituba collected in winter and spring were higher than these collected in summer and autumn.In addition, the highest value of calcium and magnesium contents were found in January and February, and the relatively higher contents of Zinc, manganese, boron and iron were found on winter. The contents of phosphorus and potassium decreased from March to January of the nest year. |
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