Screening Of Resveratrol-producing Endophyte Of XinJiang Wine Grape And Studying On Fermentation Process

Endophytes are one kind of large,ubiquitous fungus group,Which can produce plentiful bioactivesubstances,most of them having the same or similar structure with its host plants. That provides animportant way to development and utilization for the discovery of new compounds,the development ofnew drugs and the protection of endangered plants. thus it becomes a new investigative hot spot.Endophytes were isolated and purified from roots,stems and leaves of the Xinjiang wine grape by tissueisolation method. We selected efficient strains that could produce resveratrol from endophytes we haveisolated, determined the taxonomic status, and optimize its fermentation process by response surface test,The results obtained are following：1,Isolation and purification of EndophytesWe collected roots,stems,leaves and other tissues and organs of wine grapes in three differentseasons of spring,summer,autumn.73endophytes were isolated include endophyticbacteria,actinomycetes and fungi.and the number of bacteria was23,actinomycetes was14, mould was24,yeast was12.and its rule is:Spring （30.14%）=summer （30.14%）<autumn （39.73%）; fruits （12.33%）<leaves （20.55%）<stem （32.88%）<root （34.25%）.2, Screening and Identification of target strainsIsolated endophytic fungi were inoculated into PDA liquid medium, and fermentated.We used ethylacetate to extract its resveratrol.By chemical color reaction,thinlayer chromatography, UV wavelengthscanning,high performace liquid detection and genetic stability experiments,we got one endophyte thatcould produce resveratr-ol,named C2J6. By morphological, physiological, biochemical and26S rDNA-ITS molecular biology techniques,we determined the taxo-nomic status, the strain was identified asAspergillus niger （partial sequence）, and established a phylogen-etic tree.3. The target strains producing resveratrol’s optimization of the process conditionsStudied on the fermentation medium and conditions of target strains producingresveratrol, Through the single factor experiment, factors and levels of effecting processing resveratrolhave been determined,: cultivation time, incubation temperature,pH, rotational speed, inoculum size,liquid volume in flask,Carbon Source, nitrogen source, trace element; Based on that,wo did fractionalfactorial design, the significant factors of effecting Fermentation Conditions were as follows: Sucrose,pH, rotational speed, Urea, manganese sulfate; Finally, through a central composite experimental,theoptimal fermentation conditions was:2.10%of sucrose,pH4.94,1.36%of urea,0.00847%of manganesesulfate,28℃of temperature, speed90rpm,20%of the liquid volume in8%, inoculum size, resveratrolyield up to this time1.965g·L^-1.