Study On The Resources Science Of Fritillaria L. From Xinjiang Uygur Autonomous Region

Fritillaria species are abundant,and there are about 140 species over the world,in which 24 species distributed in China.Fritillaria is widely used as ornamental flower.Some species,named after Beimu in China,are famous traditional medicines with medicinal effect of relieving cough and eliminating phlegm.Five types of Beimu were recorded in the "Pharmacopoeia of the People’s Republic of China",i.e."Chuan Beimu,Zhe Beimu,Hubei Beimu,Ping Beimu,Yi Beimu",in which "Chuan Beimu" and "Yi Beimu" are multi-original medicinal herbs.Source plants of these Beimu herbs involved 10 species and 1 variety.There are more researches focusing on "Chuan Beimu",while the studies of the "Yi Beimu" are relatively rare.There is 9 species of Fritillaria distributed in Xinjiang Uygur Autonomous Region.The source plants of "Yi Beimu" in Chines pharmacopoeia are only F.pallidiflora and F.walujewii.However the local people use more than the two species in the Xinjiang region as Beimu,therefore this situation may have certain impact on the safety and efficacy of Beimu herbs.The morphological characteristics,microscopic characteristics,phylogenetic relations and chemical composition were studied to investigate Fritillaria in Xinjiang.We hope by these methods to clarify the genetic relationship among species,and to provide data support for the rationality of Beimu herbs application.Combining wth literature analysis and field survey,we investigate the resources of Fritillaria species in this area.F.pallidiflora,F.walujewii,F.verticillata、F.yuminensis,F.tortifolia,F.meleagroides,F.karelinii are common in northern Xinjiang that mainly distributed in Tianshan Mountains,Yili river valleys and marginal area of Junggar Basin.The wild resources of these seven species declined sharply.Although F.pallidiflora was successfully cultivated since 1970s,the planting area and yield are often fluctuant by the market effect in recent years.In the market,F.pallidiflora is main source of "Yi Beimu".Those big bulbs of other five to six speices were mixed into Yi Beimu herbs,while their small ones were used as "Chuan Beimu" due to its higher price.F.karelinii is also more often used as ornamental flower to sell.The microscopic characteristics of leaf epidermis,pollen and bulb powder of the 7 common species and F.ussuriensis,a species distributed in northeastern China,were observed to explore relationship of the Fritillaria species distributed in Xinjiang region.The micromorphology of leaf epidermis indicated that upper epidermis without stomata of F.walujewii and F.ussuriensis differed from other species.The exinc ornamentation of pollen grain of F.karelinii belongs to psilate type,but that of the other species belongs from zono-reticulate type to pan-reticulate one.The starch granules of F.verticillata are side chain,in contrast,that of the other species are both direct chain and side chain.Micromorphology observation showed that F.verticillata and F.yuminensis have most similar in morphology and F.karelinii differes far away from the other species obviously.The phylogenetic trees of the ITS,matK and rp116 fragments by maximum likelihood(ML),maximum parsimony(MP),and Bayesian inference(BI)were constructed to study their genetic relationship.According to the phylogenetic trees,F.pallidiflora,F.walujewii,F.tortifolia,F,yuminensis and F.verticillata formed a group named Xinjiang Beimu Group.F.meleagroides clustered a group with F.ussuriensis,a species in the northeast of China,and F.karelinii named Mixed Beimu Group.The components of the species were determined by HPLC to revealed their similarities and differences.The methodological evaluation showed that the R2 of regression equationes were all over than 0.99,and the RSD values of precision,stability and repeatability were all less than 5%.The clustering analysis showed that F.karelinii was a single group,while the other species clustered together as a mixed group.In the mixed group,F.yuminensis was closer to F.pallidiflora and F.walujewii,F.verticillata,F.tortifolia,F.meleagroides and F.ussuriensis had a shorter distance.Generally the following conclusions are drawn:1.From the morphology and molecular phylogeny,F.tortifolia,F.verticillata,F.yuminensis are closer to the two source plant of Yi Beimu(F.pallidiflora and F.walujewii),while F.karelinii and F.meleagroides are far away from them.2.Basing on the genetic and chemical composition we preliminarily consider that F.karelinii has so little similarity to other species that using it as Beimu herb is questionalble.And F.yuminensis has the potential to be used as Yi beimu because of it further relationship and similar composition to F.pallidiflora.It is difficult to separate F.meleagroides,F.verticillata and F.tortifolia to F.walujewii(Yi Beimu)and F.ussuriensis(Ping Beimu)in the cluster analysis,and it therefore need to be further studies whether they all are used as congener medicinal herbs.