The Endangered Medicinal Plant Phellodendron Amurense Production Of Active Ingredients Of Endophytic Fungi

Endophytic fungi were isolated out from the stems and leaves of a rare medicinal plant Phellodendron amurense Rupr. The following studies were carried out on the basis of these fungi.By the deposition reaction with Dragendorffs agent and a special color reaction, two strains ,S6 and L2 ,were screened out. Then S6 was selected out bythiner layer chromatography(TLC) because of its higher yield of alkaloid of berberine (Ber). Through further culture and by TLC, some procedure of extraction were studied.The extract of the culture filtrate of S6 was compared with the standard BBH by TLC and their spots were scanned under UV and visible spectrum. It was found that the absorption curves of the extract was basically the same as that of the standard sample. The results of IR and HNMR was analyzed and it was supposed that the extract had some typical structure that BBH had.According to the growth rates of mycelia and growth rate of the colonies on different media, an optimum solid medium was selected;through analysising the amount of biomass and the yield of alkaloid, an optimum liquid medium was selected out;then, by changing different carbon source, nitrogen scource, pH and culture temperature, the culture conditions were selected. The optimized condiations of mycelical growth and alkaloid production are maltose, urine, pH about 7.0 and the temperature about 28℃, respectively.The fermentation conditions of S6 were then further optimized by the orthogonal experiment of L16 (4³) .The results showed that maltose 15g.L^-1, urine 4.3 g.L^-1, pH about 7.0 and the temperature about 28℃, rev 80r/min were the optimum conditions for its mycelial growth;maltose 15g.L^-1, urine 5.3 g. L^-1, pH about 7.0 and the temperature about 28℃, rev 80r/min were suitable for alkaloid production.