Phylogenetic Research Of Amanita Section Amanita Species Based On Isoxazoles And Secondary Metabolites

Poisoning caused by wild mushrooms ingestion accounts for a large proportion of food poisoning incidents in China.In recent years,neurotoxicity caused by wild mushrooms ingestion occurs frequently.The neurotoxic mushroom in the species of Amanita section Amanita contains isoxazole derivatives.However,the toxins of which has not been reported in China.In addition,the fingerprinting of secondary metabolism of Amanita section Amanita and its relationship with phylogenetic tree based on ITS has not been studied internationally.Therefore,to detect the toxins of poisonous mushrooms containing isoxazole derivatives,and to analyse the relationship of fingerprint based on toxin and related metabolic components and phylogenetic tree based on ITS has important academic and application value.In this paper the optimization on the extraction and detection of isoxazole derivatives,the detection of ibotenic acid in species of Amanita section Amanita and the relationship of phylogenetic tree based on ITS and phylogenetic tree based on secondary metabolites was carried out.The findings are as follows:1.Optimization of extraction and detection of isoxazole derivatives: the contents of secondary metabolites was higher and the extraction time was shorter by means of the ultrasound extraction method by comparing the HPLC profiles.For the mobile phases,the results showed that the formic acid aqueous solution and formic acid acetonitrile solution are the most suitable mobile phase for the detection of isoxazole derivatives and the construction of fingerprint by comparing the degree of separation and the number of HPLC peaks.2.Content of ibotenic acid: 22 samples from 12 species of Amanita section Amanita was detected.The results show all the species contain ibotenic acid,which ranged from 4.45-7.87 mg/g.3.Comparison of phylogenetic analysis based on secondary metabolites and phylogenetic analysis based on ITS: HPLC fingerprint of secondary metabolites was established by using the amantadine column the amino column,and then the clustering tree was constructed according to the fingerprint and compared with phylogenetic tree based on ITS fragments.The results show that both phylogenetic tree based on ITS and cluster tree based on secondary metabolites could divide 22 tested species into 8 groups,in which the grouping situation was basically the same,and the two developmental trees were basically consistent.Which indicated that the production of secondary metabolites of the species are associated with their gene development.