Setting Up The Culture Technology System Of Taxus Media And The Separation And Identification Of Endophytic Fungi

In the experiment,Taxus media was used as test material. We studied the best medium and culture conditions for callus induction and the Effect of plant growth regulator type and concentration on callus induction and callus culture of subculture, the Effect of the concentration of anti-browning substances on callus browning, the Effect of different subculture time on the browning of callus, the separation and identification of endophytic fungi of Taxus media cv.Hicksii. Through this test, the purpose is to comprehensively study the main factors affecting each stage and the optimum medium, and the optimal technical parameters were selected to gradually establish a tissue technology system for Taxus . Through the separation and identification of endophytic fungi of Taxus media cv.Hicksii, we aimed to find endophytic fungi which product Taxol, and further to provide certain theoretical basis for production of Taxol using microorganism fermentation.Study made by the following results1. The results of the study on best medium for callus induction and culture conditions were as follows. In MS, B₅, TA medium, light and dark culture ,on the basis of single factor experiments, selected the best medium and most suitable culture condition, and B₅ medium was the best medium, and the best culture condition was the dark culture.2. The optimum medium recipe of callus induction: in the B₅ medium and dark condition, studying the Influence of the different plants growth regulating substances and concentration, the result was that the optimum medium recipe of B₅ medium was B₅ + 0.25 mg/L 2, 4-D + 1.5 mg/L NAA + 0.5 mg/L KT.3. The optimum medium recipe of callus proliferation subculture: on the basis of optimum medium recipe, improving plant growth regulating substances to study the influence of different concentration on the callus proliferation, the best recipe was that B₅+0.5 mg/L 2,4-D+2.0 mg/L NAA+1.5 mg/L.4. Different anti-browning substances and their concentrations on medium formula reducing callus Browning: based on B₅ +0.5 mg / L 2,4-D +2.0 mg / L NAA +1.5 mg / L, different type and concentration of anti-browning substances were added to relieve the callus browning. Better to inhibit the callus browning is the optimum medium, which is B₅ + activated carbon 0.4 g / L + hydrolyzed milk protein 0.5 g / L + PVP 0.3 g / L. 5. Different subculture time on the browning of callus: Add the most appropriate anti-browning substances and their concentration, we found different subculture time on callus browning is also influential. On the ten day, we select the most favorable subculture which can better inhibit callus browning.6. From the bark of Taxus five isolated strains of endophytic fungi,forty The identification of five belong to different genera, the five strains fermentation, the concentrated extract, two strains detected by TLC Rf = 0.40 and Rf values consistent with the paclitaxel. No.two can be determined basically Taxus strains of endophytic fungi producing taxol.