Research On The Endophyte Screening Of High-yield Alkaloid Strains And Its Pharmacological Activity Of Fritillaria Ussuriensis Maxim

Fritillaria medicines can be divided into five types according to their original plant species, namely Chuanbeimu, Zhebeimu, Pingbeimu, Yibeimu and Hubeibeimu. All of the drugs are from the perennial herbs of the Liliaceae family, their bulbs as one kind of the traditional Chines medicines, have the effect of cleaning the heat and dispelling phlegm, nourishing the lung and relieving the cough, generally considered the alkaloids as the active ingredients. Endophyte is a research hotspot in recent years; it has been found that endophyte has the capable of producing similar chemical componets to the host plant. Therefore, it has higher theoretical and practical significance to use the Pingbeimu endophytes being found, to research their abilities in producing secondary metabolites, and to study the pharmacological activity of these products, to get rid of the constraints of environmental conditions in the field, to carry out its industrial production.In this paper, our laboratory isolated Pingbeimu fungi as experimental materials, screening high-yielding strains, then optimizating culture conditions, finaly to carry out the pharmacological activities. The main conclusions are:(1) The fermentation products of the existing 10 strains of endophytic fungi(P1,P2,P3,P4,P5,P6,P7,P8,P9,P10) were screened by thin-layer chromatography identification, strain P7 result of the determination of the strain is capable of a higher level Fritillaria alkali.(2) Based on the factors of solid culture medium, liquid medium, carbon source, nitrogen source and temperature, the single factor optimization of P7 test was carried out, the optimal conditions is the potato culture medium; and glucose as carbon source, urea as nitrogen source, the culture temperature is 26℃, the fungi can grow well, and reach the maximum alkaloid content. Furtherlay, a orthogonal experiment was desgined with four factors and four levels, according to the data analysis the optimum fermentation conditions of P7 is identified as the followings: 60 g / L glucose, 0.5 g / L urea, 150 ml liquid volume, pH = 7.0.(3) The ten endophytic fungi fermentation broths and mycelia were use in the study of anti-tumor, anti-monoamine oxidase and acetylcholinesterase activity. The rusults show that P7 strain has a certain anti-human cervical carcinoma Hela cell activity in bacteria extract of mycelium, and the activity may reach 18.4% ± 0.4%, higher than 30% of the fermentation broth extract; P4 strain’s anti-monoamine oxidase activity was highest in the extract of mycelium, the activity may reach 63.7% ± 0.5%, higher than 35% of the fermentation broth extract: P3 strain’s anti-acetylcholinesterase activity is the highest in extracts of mycelia, the activity may reach 69.1% ± 0.5%, higher than the fermentation broth extract.