Endophytes From Phlegmariurus Carinatus And Their Second Metabolites Inhibiting AChE Activity

Phlegmariurus carinatus, one of ornamental plant growing on rock or attaching on tree in nature usually is applied in gardening as hanging plants.Now further research about this plant found that Huperzine A,extracted from Phlegmariurus carinatus, has shown promise in Alzheimer's Disease (AD) as an inhibiting AchE agent. Unfortunately, the supply issue is complicated by Phlegmariurus carinatus which is a slow-growing fern .The content of huperzine A in that is very low.Recent reseaches have been showing that there are many taxa of endophytes existing in their healthy and asymptomatical host plants. These endophytes were ubiquitous and can produce various second metabolites which have different biological activity and have potencial applied value in agriculture and medical.Our reseach set out to isolate endophytes from Phlegmariurus carinatus (Desv) ching and investigate their distribution and genus Taxus of these endophytes. Meantime, We aimed at disclosing whether endophytes inside Phlegmariurus carinatus (Desv) ching can produce substances which have inhibiting AchE activity.A total of 56 endophytes were isolates and identified by their morphological characters. The endophyte assemblages consisted of 41 ascomycete and 9 zygomycete and 6 sterile fungi. Most of them live in stem of their host plant,others in leaves, scarece in roots. The ferment solution of these endophytes was evaluated in vitro for inhibiting activity to AchE. Out of 56 endophytes investigated, 13 can produce unknown substance that were inhibiting to AchE in vitro and accumulating after trophophase. The rude alkoids from one of endophytes ferment solution have ability to inhibiting AchE which IC50 is 410μg/L,equal to the alkoids from Huperzia serrata (Thunb.) Tre v.. The condition of optimum culture for K11 strain were as follows: the best of medium was PDA and the suitable culture temperature for Kll strain in was 26℃±1℃. And the speed of shaking incubator for culture K11 strain was 120rpm or more than 120rpm. Further investing analysis of substance inhibiting to AchE by TLC and HPLC show the inhibiting substance were alkoids of huperzine.