Diversity,Phylogeny And Anti-Lung Cancer Cell Line A549 Activity Of Endophytes Associated With Polyporus Umbellatus

Cancer has become one of the most threatening diseases in the world due to its high incidence rate.Finding highly effective anti-cancer drugs is a hot topic in medical research.Many studies have shown that metabolites of some endophytic fungi and bacteria isolated from medicinal plants have a moderate anticancer activity.This paper takes Polyporus umbellatus as the research object,and systematically studies the diversity and anticancer activity of endophytes isolated from sclerotia tissue.The endophytic strains with medicinal value were screened out by detecting the anticancer activity of the crude extracts in the fermentation broth of sclerotia endophytes.The results are as follow:1.18 strains of endophytic fungi were isolated from the sclerotia of Polyporus umbellatus.Through spore morphology,colony morphology,and sequencing of the ITS region,the results showed that 18 endophytic fungi belonged to seven different genera,seven of which were Trichoderma sp.,four Nectriaceae sp.strains,two Scytalidium lignicola strains,one Mariannaea camptospora strain,one Mariannaea elegans strain,one Fusarium sp.strain,one Penicillium sp.strain and one Aspergillus sp.strain.Therefore,Trichoderma sp.is a predominant group in sclerotia tissue of Polyporus umbellatus,accounting for 38.9% of the total number of endophytic fungi.2.86 strains of endophytic bacteria were isolated from the sclerotia of Polyporus umbellatus.The strains were identified by colony morphology and sequencing of the 16 SrDNA regions.The results showed that these strains belong to 5 different genera.60 of which were Bacillus sp.,15 strains were Brevibacterium sp.,2 strains were Arthrobacter sp.,3 strains were sporosarcina,and 6 strains were Paenibacillus sp.Therefore,Bacillus sp.belonged to the dominant bacteria in the sclerotia of Polyporus umbellatus,accounting for 69.8% of the total number of endophytic bacteria.3.Using the MTT assay and regarding human lung cancer cell line A549 as the indicator cell,the crude extract of 18 endophytic fungi and 86 endophytic bacteria were used as the tested substances for anticancer activity.The test results showed that 4 strains of fungi showed moderate inhibition of tumor cell proliferation,which accounted for about 22.2% of the total number of tested strains,2 strains of bacteria had the effect of inhibiting the proliferation of tumor cells,accounting for about 2.3% of the total number of tested strains.Among them,the crude extracts of endophytes MG515066,MG515074,MG515075,MG709163 and MG709183 had a significant inhibitory effect on cell proliferation,respectively.4.The UPLC-MS assay of crude extracts from endophytes(MG515065,MG515066,MG515074,MG515075,MG709163,and MG709183)showed that the crude extracts of fermentation broths contained some of the same components.It is speculated that the crude extract of endophytic fermentation broth inhibited the proliferation of lung cancer cell line A549 may be the result of the common components.