Study On Strain Fermentative Secondary Metabolites And Anti-fungal Activity Of Cortinarius Rufo-olivaceus(Pere.)Fr.

Cortinarius rufo-olivaceus?Pere.?Fr.is a kind of rare wild edible macro fungi resource which mainly distribute in the Helan Mountain in Ningxia Hui Autonomous Region and Inner Mongolia Autonomous Region in China.Many researches have reported that the secondary metabolites of it exhibit abundant bioactivities.However,the macro fungi species that born on Helan Mountain are abundant,and the morphological characteristics of these macro fungi are similar.Classification of these fungi based on morphological characteristics is not accurate enough.And the research on the secondary metabolites and bioactivities of C.rufo-olivaceus is currently focused on its fruiting bodies.In this experiment,two methods of phenotypic taxonomy and genotypic taxonomy were used to conduct systematic taxonomic research and identification of C.rufo-olivaceus.We isolated the strain of C.rufo-olivaceus,and selects the most suitable culture medium by comparing fermentative products of C.rufo-olivaceus for large-scale fermentation.Filtration,extraction and various chromatographic techniques were used to separate and purify the obtained fermentative products,and various spectrometry techniques were used to identify the structure of the obtained monomer compounds.The similarities and differences between the secondary metabolites of the C.rufo-olivaceus fruiting body and fermentative products were compared at the same time,and the crude extract of the fermentative products was tested for anti-fungal activity,and the following results were finally obtained:1.By comparing the morphological characteristics of C.rufo-olivaceus described in the literature confirmed the collected macro fungi was C.rufo-olivaceus,The fungus was further identified by ITS sequencing which showed that it had 99.57%homology with Cortinarius rufo-olivaceus TUB 011463,.2.The oat juice medium was finally selected as the most suitable medium for C.rufo-olivaceus by comparing the chemical composition of the fermentative products from different medium,and the secondary metabolites of C.rufo-olivaceus in this medium were the most abundant,and it is also most similar to the secondary metabolites of fruiting bodies.3.By separating and purifying the ethyl acetate phase of the fermentation product of C.rufo-olivaceus,18 compounds were isolated and the structure of 15 compounds was identified,including 3 steroids and 12 anthraquinones.These 15 compounds are all obtained from C.rufo-olivaceus by fermentation for the first time,of which compound Cr-14 is a new natural product.Cr-1,Cr-2,Cr-4,Cr-10,Cr-12 and Cr-13have also been reported in the fruiting bodies of C.rufo-olivaceus,However,the compounds Cr-3,Cr-5,Cr-9 and Cr-15 have not been reported in the secondary metabolites of the C.rufo-olivaceus fruiting body,which demonstrate that not only some of the same chemical components as the fruiting body but also new compounds can be produced by fermentation.4.The antifungal activity test of the crude extract of C.rufo-olivaceus against plant pathogens showed that the BuOH as well H₂O extract of C.rufo-olivaceus had weak antifungi activity against Botrytis cinerea and Magnaporthe grisea,but the ethyl acetate phase shows significant antibacterial activity.In summary,the strain of C.rufo-olivaceus was isolated from the fruiting bodies,and the ITS sequencing result proved that the strain had 99.57%homology with Cortinarius rufo-olivaceus TUB 011463,which further confirmed its taxonomic status.Taking this strain as the research object,18 monomer compounds were isolated from the oatmeal medium fermentative products,6 of which have also been reported in fruit body extracts,it provides a theoretical basis for achieving the quantitative production of active compound by fermentation.However,the biological activities of the fermentative products needs further study.