Study On The Diversity Of Endophytic Fungi, Fermentation Products And Changes In The Drying Process Of Licorice

Endophytic fungi have long been coexistence with host plants in the form of parasitism or symbiosis,especially fungi born in some medicinal plants.In the process of co evolution with the host plants,it participates in the metabolism of medicinal plants or the synthesis process of some kind of active compounds,and can produce the same or similar compounds.An important source of new natural products.Screening endophytic fungi from Glycyrrhiza uralensis and other effective components,and then using fermentation culture to produce effective components of Glycyrrhiza uralensis,may be a new choice.First,to select the suitable culture conditions,optimize the surface sterilization conditions and select the best Glycyrrhiza material three aspects,to provide a basis for better isolation of Endophytic Fungi from licorice root.Secondly,we studied the diversity of Endophytic Fungi from licorice from different habitats,and constructed the endophytic fungus Library of Glycyrrhiza uralensis.The samples of Glycyrrhiza Glycyrrhizae were collected from different habitats,and a large number of endophytic fungi were obtained after separation and purification.The species of endophytic fungi were identified by morphology and ITS sequence,and the endophytic fungi diversity in different areas of Glycyrrhiza was studied.Thirdly,the effective components in the fermentation products of Endophytic Fungi from Glycyrrhiza uralensis were detected by HPLC screening and UPLC-MS reexamination,and the effective components of Glycyrrhiza,glycyrrhizin,glycyrrhizin,glycyrrhizin and ISO glycyrrhizin were selected.Finally,we selected the different time points of licorice drying process to detect the species and quantity of endophytic fungi in Glycyrrhiza uralensis,and to study the change rule.The main results of this study are as follows:(1)The cultivation of endophytic fungi in Glycyrrhiza uralensis can be cultivated with PDA,PDB medium and green streptomycin mixed solution to avoid bacterial contamination.The addition of a concentration of 50 to g/m-100 g/mL can eliminate bacteria without affecting the growth of fungi.The best surface sterilization conditions are:soak 2min in 75%ethanol,rinse 1min with sterile water,soak 5min in 5%sodium hypochlorite solution,rinse lmin with sterile water,rinse 30 S with 75%ethanol,rinse lmin with sterile water,and finally dry out surface moisture with sterile filter paper.If we want to get as many endophytic fungi as possible,we should select three years' raw material for licorice cultivation.(2)There are 438 endophytic fungi which were obtained from 129 licorice samples from 5 regions of Gansu,Ningxia,Inner Mongolia,Xinjiang and Beijing,and 438 endophytic fungi were isolated.After morphological identification,microscopic observation and ITS identification of fungi,26 species were classified into 5 orders,8 genera and 26 species,including Aspergillus,Penicillium,Penicillium and Fusarium Fusrium.Alternaria,Alternaria,Neocosmospora,Uncultured Glomerales,Naganishia Albida,and Uncultured fungus;among them,there are 10 species identified,of which 8 are isolated and identified for the first time in Glycyrrhiza;Bionectriaceae,Neocosmospora,Glomerales,and Naganishia are in China for the first time.The region is separated from licorice.Fusarium,Aspergillus and Penicillium are the dominant bacteria in licorice.There were differences in the isolation rate of Endophytic Fungi from different habitats:Ningxia(85.5%)>Gansu(63.2%)>Xinjiang(62.8%)>Inner Mongolia(55.5%)>Beijing(37.4%),and the real estate area was larger than non real estate area.The distribution and distribution of Endophytic Fungi of Glycyrrhiza uralensis in different producing areas are different:Fusarium,Aspergillus,Penicillium and Alternaria are dominant species.In Gansu,Xinjiang and Inner Mongolia,Fusarium and Aspergillus are dominant in the endophytic fungi of licorice,and Fusarium in Ningxia licorice is the dominant bacteria,and the dominant fungus of Endophytic Fungi of licorice in Beijing area is Fusarium.Fusarium species are common dominant bacteria in different producing areas.The diversity of endophytic fungi in different habitats was different:the diversity index Gansu(4.81)>Xinjiang(2.40)>Ningxia(2.19)>Inner Mongolia(2.10)>Beijing(1.70),and the real estate area was larger than the non real estate area.(3)After the fermentation of endophytic fungi obtained from Glycyrrhiza Glycyrrhiza,an endophytic fungus producing glycyrrhizic acid was obtained by HPLC screening UPLC-MS,and ZK-F10 has been identified as a fungus of the genus agarospora.(4)In the drying process of Glycyrrhiza uralensis,the number and the number of endophytic fungi increased rapidly,and then increased slightly.