Intraspecies Variance Pattern And Their Relationship With Hupa Content In Huperzia Serrata From Zhejiang And Its Adjacent Area

Huperzia serrata (Thunb.) Trev., an important Chinese traditional medicine plant, belongs to Huperziaceae, Lycopodiales, Lycophytina, Pteridophyta. Two genera and about 50 species have been reported from Chinese Haperziaceae. Most species of the family Hupersziaceae contain Lycopodium alkaloids, among them, HupA has been proved to be medically effective to Aezheimer's disease, with the highest content in the species H. serrata. Ecologically, Huperzia serrata is earthy, epiphytic herb or grows on bryophyte layer in the forest, usually in shady and humidity environments.Eight populations of H. serrata collected from Pan'an Dapanshan, Jinhua Beishan, Qingyuan Baishanzu, Suichang Jiulongshan, Suichang Daxikeng, Lin'an Xitianmushan, Fujian Wuyishan as experimental materials, several analytic and fingerprint technologies, including paraffin-section, scanning electron microscope, RAPD, IR and HPLC were applied to collected comparative indices, and Principal component analyze (PCA) and Detrended Correspondence Analysis (DCA) were applied to analyze these obtained data to show the intraspecies variaions of the species H. serrata in Zhejiang and its adjacent area. The main results are as follows:1. The content of HupA in 8 populations of H. serata and 2 populations of Phlegmariurus mingchegensis were determined by HPLC using Waters 1525-2487. The results showed that: (1) HupA was found in all samples, but its content highly varied, with the highest content in the population collected from Dapanshan of Pan'an (average content 0.475mg·g^-1 in stems and leaves ) , the HupA content of the other populations varied from 0.02 mg·g^-1-0.11 mg·g^-1; (2) for the population collected from the same site, the content of HupA in the stems was different from those in the leaves, in the populations from Beishan and Baishanzu, their leaves contained higher content of HupA their stems, others were on the contrary; (3) determination also showed that the HupA content in P. mingchegensis from southern Zhejiang, with as high as 0.675 mg·g^-1, was much higher than most samples of Herrata serrata. Therefore, Phlegmariurus mingchegensis population from Suichang county and H. serrata Pan'an population might be two good genetic resources for cultivating and medicinal resources for exacting HupA medicine, andPan'an maybe a suitable area for cultivating H. serrata.2. Investigation of the water content, organic matter content, conductivity, pH and the content of 11 metal elements in the soils where H. serrata grew shows: (1) Huperzia serrata grows very slowly, mostly populations are discovered in dankness soil of the hill country or the close woods or the vale at altitude 350-1600m, with 90% or more canopy density, over 1500 mm rainfall per year and over 78% relative humidity; (2) The soils where H. serrata grew was characterized by having 10% - 30% water content, 4.57 - 5.31 pH, 0.061 - OJSSms-cm"1 conductivity, and 6.18% -9.75% organic matter content; (3) Huperzia serrata demands more K> Ca> Zn and Na than other elements, and easily accumulated Pb> Cu and Cd. Therefore, the balance of element content should be paid more attention in cultivation. The DCA scatter plot based on the soil indices showed that the environmental variables related to Pan'an population are distinctively different from the other sites, revealing the special environmental conditions for the population of H. serrata with higher HupA content to grow.3. The method of Furrier-transform Infrared Spectrometer with OMNI collector was applied to obtain the infrared spectra of 17 leaf samples including H. serrata, and its five closely related species (//. sutchueniana, Phlegmariurus mingchegensis, Lycopodium japonicum, Selaginella doederleinii, Selaginella heterostachys). Based on the indices of wave number-absorbance (666-3145 cm-1), the differences of the 17 infrared spectra were compared by the method of Principal Component Analysis (PCA). The results showed that there is good correspondence between the position relationship of PCA three-dimensional plot of the samples based on the indices of wave number-absorbance of FT-IR spectra and their phenetic relationship. Therefore, the infrared spectra could be applied to identify the samples of H. serrata and its closely related species. Results also showed that the infrared spectra of Pan'an sample with higher HupA content are distinctly different from the others.4. The exacting solutions of eight samples of H. serrata and two samples of P. mingchegensis from zhejiang and Fujian province were scanned by UV. DCA on the UV spectra of the solutions of these 10 samples showed: (1) the similarity of chemical composition in these populations could be revealed in the DCA scatter plot; (2) the speciality of the population from Pan'an with the highest content of HupA could also be...