Screening The Traditional Chinese Medicines By Metabolites Biosynthetic Genes Of Endophytes

OBJECTIVE To screen the specialized metabolites biosynthetic genes of endophytes from Tradiational Chinese medicines plant with anti-cancer activity, and scree the secondary metabolites of endophytes with anti-cancer activity. METHODS Accroading to the experiences of the effects through documented trials or reported anticancer activity, collected34TCM plants prepared for total DNA extraction; use PCR to selectively amplify endophyte metabolites biosynthetic genes; then collected the positive plants, isolated and cultured their endophytes. Phylogenetic analysis of the isolated endophytes, based on the16s rRNA gene of bacteria and the18s rRNA gene of fungi, revealed diverse bacterial and fungal genera. The endophytes have the PKS or NRPS biosynthesis genes were selected for detection the anti-cancer activity of their secondary metabolites. RESULTS7plants were found contained the specialized metabolites biosynthetic genes of endophytes, then49endophytes were isolated; include13bacterial strains (26.5%) and36fungal strains (73.5%). The identification of49endophytes by rRNA assay indicated that the fungal strains were distribute in at least13genera and a lot of the strains were Penicilliu spp.(19.4%) and Phomopsis(25%), the baterial strains were distribute in at least13genera. Successful screened23fungal strains contained PKS gene fragments(63.9%),13bacterial strains contained PKS gene fragments (69.2%),1fungal strain contained NRPS gene fragments and1bacterial strain contained NRPS gene fragments.6bioactive endophytic fungi were selected to study their secondary metabolites. Only3endophytic fungi showed bioactivity on tumour cell. CONCLUSION This study demonstrate that TCM plants and especially their endopnytes could produce bioactive compounds, and that genetic screening targeting PKS and NRPS biosynthesis genes provides a suitable platform to highlight endophyte of relevance to drug discovery investigations.