Isolation,Identification And Antimicrobial Activity Of Endophytic Fungi From Aconitum Leucostomum

Is a perennial herb of Aconitum leucostomum Ranunculaceae Aconitum plants,its roots can be used for the treatment of the disease of the body and limb paralysis,joint swelling pain,rheumatism and cold pain and diarrhea.Aconitine is the main secondary metabolite of diphtheria,aconite root and aconite.It has good anti-inflammatory,anti-tumor and analgesic effects.Studies have shown that endophytic fungi can produce the same or similar active components as the secondary metabolites of host plants,so the endophytic fungus of diphtheria Aconitum can be used as a potential resource for screening new drugs.This paper obtained 70 strains have different morphological characteristics of endophytic fungi isolated from Yili County of Zhaosu wild Aconitum leucostomum root using tissue isolation method,through the separation and determination of fermentation broth on Escherichia coli,Bacillus subtilis,Staphylococcus aureus and Candida albicans antibacterial effect of disk diffusion method.The results showed that 20 strains of endophytic fungi have the antibacterial activity of different levels,including 3 strains of endophytic fungi had strong antibacterial activity against 4 indicator bacteria,1 strains of fungi have strong antibacterial activity on 3 kinds of indicator bacteria,2 strains of endophytic fungi have the antibacterial activity of the middle level of the 3 kinds of indicator bacteria,4 strains of endophytic fungi have antibacterial activity on 2 kinds of medium level indicator bacteria,10 strains of endophytic fungi had antibacterial activity on 1 kinds of medium level indicator.The results showed that the 5 strains of Aconitum endophytic fungi exhibited strong antibacterial activity,and the antibacterial activities of strains YLAC-3,YLAC-20 and YLAC-69 were broad.In order to determine the taxonomic status of endophytic fungi,the first of 20 strains with antibacterial activity of endophytic fungi colony,mycelium and conidia morphology were observed by optical microscopy,and then use PCR from endophytic fungi genomic DNA amplified rRNA gene fragment of ITS,and a phylogenetic tree using Clastal X8.1 and Mega 3 the software construction of ITS sequence.The identification results showed that 3 strains of Aspergillus strains,5 strains of bacteria basidiomycotina,6 strains were Aureobasidium,1 strains of Simplicillium lanosoniveum,1 strains were Coprinellus domesticus,1 strains were Penicillium sp..Thin layer chromatography(TLC)and high performance liquid chromatography(HPLC)were used to detect aconitine in fungal fermentation broth and mycelia extracts.The antibacterial activity of 20 strains of endophytic fungi in only 1 strains of Aspergillus tubingensis YLAC-3 can produce aconitine,namely in strain YLAC-3 fermentation broth and mycelia extract appeared with aconitine standard absorption peaks similar.Conclusion: 20 strains with antimicrobial activity of endophytic fungi isolated from the roots of Aconitum leucostomum,their fermentation liquid on indicator bacteria showed strong antibacterial activity,and confirmed that Aconitum leucostomum endophytic fungi is a new resource for new antimicrobial anticancer active substances.