Contents Of Active Ingredients In Huperziaceae And Molecular Systematics In Phlegmariurus From China

Objective: Toextract and determine total Huperzine A and Huperzine B in Huperzioideae and explore the methods of sustainable use of this subfamily.At the same time,to explore the phylogenetic relationship and genetic structure of this subfamily,and analyze the genetic diversity.Methods: In this paper,Huperzine A and Huperzine B content in Huperziaceae was determined by HPLC,and one-way ANOVA was used to compare differences in contents.We screened 6 pairs cp DNA primers which had been used to amplify 14 species of this subfamily which from China.The sequence results had been used to constructed phylogenetic tree by maximum likelihood method and Bayesian method after comparison and correction.Results: 1?In this study,68 samples which contained 20 species had been tested,and Huperzine A was found in 65 samples,Huperzine B was found in 58 samples.The content of Huperzine A in Phlegmariurus hamiltonii was the highest,which reached up to 0.18487%,the second most was in P.cryptomerianus which reached up to 0.15116%,and it was extremely rare in Huperzia selago and P.squarrosus.The content ofHuperzine B in P.gracilicaulis was the highest,which reached up to0.03536%.The higher levels of it in H.serrata which reached up to0.02121 %,2?There is a large variation in cp DNA,and the molecular phylogenetic tree of Phlegmariurus from China based on six cp DNA regions revealed that Phlegmariurus is monophyletic,four groups can be divided,P.cryptomerianus,P.minchegensis,P.austrosinicus and P.petiolatus are the first group,P.fargesii and P.yunnanensis are the second group,P.guangdongensis,P.squarrosus,P.shangsiensis,P.henryi and P.fordii are the third group,P.carinatus,P.phlegmaria and P.salvinioides are the fourth group?Conclusions: 1?In this study,Huperzine A and Huperzine B were found in 20 species of Huperziaceae.There were differences in the contents from species to species in this subfamily.The wild resources of Huperziaceae were scarce and should be protect.The advantage species which obtain more Huperzine can be selected based on this study,and the sustainable utilization resources can be obtained by means of tissue culture and artificial planting.2?The molecular phylogenetic results is not supported the old groups which based on morphological study.The taxonomic relationship should be rediscussed in combination with molecular phylogeny and morphology.The14 species of Phlegmariurus from China are independent taxa,what has been used to determine the protection level of endangered species and to determinethe priority protection units.