Genetic Diversity Analysis And Biotransformation Research Among Endophytic Fungi From Taxus

Taxus, a genus of yews, known as the "living fossil plant", is a small coniferous tree or shrub in the yew family Taxaceae. Endophytic fungi are microorganisms living within the tissues of host plants without causing discernible manifestation of disease, and have proven to be one of the important sources of a variety of structurally novel active secondary metabolites with anticancer, antimicrobial and other biological activities. Endophytic fungi of Taxus have attracted great attention on the areas of medicine research.20 endophytic fungi were classified by morphological identification method. The results showed that:20 endophytic fungi were classified four groups by traditional morphological identification, including Aspergillus, Nigrospora, Penicillium, Trichoderma. The relevance of secondary metabolites of endophytic fungi was studied combined with TLC, UV and HPLC techniques in this research. The results showed EFTâ…¤-01, EFTâ… -03 have the ability to yield paclitaxel or paclitaxel analogues. It suggested that metabolites of strains from the same genus have some relevance.The genetic diversity of 20 endophytic fungi was analyzed by sequence-related amplified polymorphism (SRAP) technique. SRAP analysis was done with 24 pairs of primers. The data were analyzed by NTSYS-pc 2.1 software. All strains were uniquely distinguished by 24 pairs of SRAP primers with 584 bands and 446 polymorphism bands which generated 76.4% polymorphic ratio, with the size ranging from 100 to 2000 bp, and clustered into four major groups at GS=0.601 and further separated into nine sub-groups at GS=0.921 by UPGMA cluster. The results suggested that endophytic fungi from taxus possessed highly genetic diversity. It suggested that SRAP could really reveal the genetic diversity of endophytic fungi strains from Taxus, and also could be used as a method for genetic diversity analysis among endophytic fungi from Taxus. This study provides foundation for further study of the taxol-producing endophytic fungi based on our analysis.The microbial transformation of raisins extract and absinthe oil by endophytic fungi isolated from the taxus was investigated to obtain some new pharmaceutical active ingredients. Screening test for the microbial transformation was carried out using TLC and UPLC and GC-MS techniques. The results showed that endophytic fungi EFTâ…¤-01 can produce three new compounds, including 2,2-Dimethyloctanol, 2,4-diisopropyl-1,1-dimethyl-Cyclohexane and cyclic 1,2-ethanediyl acetal, (5.alpha.)-Cholestan-3-one, with the substrate raisins ethanol extract. It also can produce new terpenoids, including (-)-(Z)-Myrtanyl acetate, delta.-Elemene, delta.-Cadinene and beta.-Farnesene, with the substrate absinthe oil.