| "Beimu"(Bulbus Fritillariae), a kind of well-known traditional Chinese medicine (TCM), have a long meicinal history since it was firstly recorded in "Shen Nong Ben Cao Jing"(Eastern Han Dynasty, A.D.20-225). Since then, more and more Fritillaria species were introduced into TCM during2000years history owing to their similar medicinal value."Chinese Pharmacopoeia (CP)"(2010edition) recorded five kinds of "Beimu", which were from11source Fritillaria species. Because of high price, the quality and authentic source of "Beimu" were often quite questionalbe in some TCM market. Some non-Pharmacopoeia species often were sold as authentic "Beimu". Meanwhile, there are disagreement in taxonomy and systematics of genus Fritillaria."Flora Republicae Popularis Sinicae"(1980) recorded20species and2varieties, but the numbers of Fritillaria species and varieties subsequently published were more than100."Flora of China (FOC)"(2000) revised this genus and recorded24species and2varieties. Although this genus was revised several times, but there still were contradiction between CP and FOC. This research attempts to clarify the taxonomic and systematic relationship among parts species of Fritillaria, especially focusing on those species distributed the middle and lower reaches of the Yangtze River, based on micro-morphology, morphological traits, molecular characters. Additionally, we discussed the plant distribution, resource situation and the usage of Fritillaria herbs in the medicinal markets.We comprehensively investigated the resources of Fritillaria thunbergii (also called Zhe Beimu in Chinese) by literature review and field survey. The history, distribution, chemical components of "Zhe Beimu" were summarized by referring to literatures. We found that "Zhe Beimu" described firstly in "Ben Cao Jing Ji Zhu"(A.D.480-498), and it has the longest medicinal history among all Fritillaria species. In the field survey, we interviewed farmers, businessmen, enterprises and members of administrative departments in order to get those information and data about the cultivation area, cultivation measures, yields and sales. In laboratory we observed the leaf epidermis, transverse section of leaves, and powder of bulbs of Fritillaria species by tissue section, light microscopy and scan electronic microscopy. We found that the anticlinal wall of epidermic cell and starch striations of bulbs of F. hupehensis is different from others. The epidermic cell shape and the anticlinal wall of F. ebeiensis is clearly different from F. anhuiensis. The anticlinal wall of F. puqiensis is similar with F. hupehensis. The leaf epidermis cells shape, starches and crystals of calcium oxalate are similar for F. anhuiensis, F. monantha and F. thunbergii. The wax ornamentations of the leaf surface of F. cirrhosa are more intensive than other species. Crystals of calcium oxalate in bulbs of F. pallidiflora, F. ussuriensis is visually bigger than other species.Cladistics method with28morphological traits was conducted to reconstruct MP tree of the Fritillaria species distributed along the middle and lower reaches of the Yangtze River. The results showed that F. thunbergii has the furthest distance from other ones, followed by F. hupehensis. The samples from F. anhuiensis and F. monantha clustered together each other. F. hupehensis has a short pedicel (c.v.1-2cm), and the cirrose apex of bract, whorled leaves. The three traits of F. hupehensis is steady and is distinct from F. monantha.The sequences of ITS, matK and rpll6were combined to reconstruct the phylogenetic trees (MP and NJ) of the Fritillaria distributed along the middle and lower reaches of the Yangtze River. We found that F. ebeiensis is furthest from other ones, then are F. hupehensis and F. puqiensis. The samples of F. anhuiensis, F. thunbergii and F. montha that were from difference places were clustered together respectively firstly, and then together all. We also reconstructed the phylogenetic trees (MP and NJ) of those species which are source of five kind of Bulbus Fritillariae in CP based on the ITS sequence. The results revealed that the source species of "Chuan Beimu","Yi Beimu" and "Ping Beimu" from difference places clustered together respectively, and the species from the middle and lower reaches of the Yangtze River got together.To sum up, the study analyzed the systematic relationship among part species of Fritillaria based on micro-morphology, morphological traits, molecular systematics. We found that the relationship between F. hupehensis and F. monantha is separated, and the similar situation are also exist in relationship of F. ebeiensis and F. anhuiensis, F. cirrhosa and F. anhuiensis, F. ussuriensis. The results of the study do not support the viewpoint of FOC, that is, combining F. hupehensis and F. monantha as one speices, F. ebeiensis and F. anhuiensis as one species. Meanwhile, we concluded that the mixed use of bulbs of F. anhuienesis and F. ussuriensis as substitutes of "Chuan Beimu" is not reasonable. |
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