Research On Isolation And Screening Of Endophytic Taxol-producing Fungi Isolated From Taxus Media And The Identification Of The Taxol High-yield Strain

In this paper,the standard method was used to isolate,purify and fermentate the endophytic fungus from Taxus media,growing in Guanzhong,Shaanxi.TLC method as primary screening measure and HPLC method as the re-screening measure were used to identify the taxol,which extracted from the fermentation of endophytic taxol-producing fungus.LC/MS/MS was used to confirm the structure of the taxol of fermentation.The high-yield endophytic fungus was precisely classified through the morphological characteristics and the molecular phylogenetics.Through the research,this paper got the following conclusion:1.26 endophytic fungus isolates were obtained by the standard method from different tissues of Taxus Media,of which 12 strains were from the root,11 strains were from sterm,1 strain were from the leaves and 2 strains were from the bark.2.8 endophytic fungus isolates,1?.3#?6#?11??15#?17#?21??26#,might be capable of producing taxol,identified by the primary screening measure,TLC.Among the strains,only 3 strains,11#?17#?and 26#,were verified to have the capability to produce the taxol with 257.92 ?g/L,428.16 ?g/L and 696.24 ?g/L by HPLC method.LC/MS/MS method was used to confirm the structure of the taxol which fermentated by the strain 26#.3.The traditional morphological characteristics,observed by the colonial morphological traits,optical microscope and scanning tunneling microscope,and molecular phylogenetics,inferred from the 18S rDNA sequences and the internal transcribed spacer region including the 5.8S rDNA(ITS)sequences,were used to classify and identify the strain 26#.The strain was classified into Aspergillus fumigatus and named Aspergillus fumigatus TMS-26.This study is the first report of the endophytic taxol-producing Aspergillus fumigatus isolated from Taxus media,which has enriched the population resource of endophytic taxol-producing fungi.It has provided a good original strain for the transformation useing mutagenesis and genetic engineering methods to build a stable high-yield production strain and the increase of production useing the optimization of fermentation conditions.