Functional Study Of The Shikimate Pathway AroF Gene In Aspergillus Coronatus

Aspergillus cristatus is the dominant strain in the blooming process of Fuzhuan tea.It can change the quality of Fuzhuan tea by producing a variety of metabolites.The early analysis of the genome of Aspergillus cristatus showed that there are two pathways in the organism to produce shikimic acid.One pathway is erythrose-4-phosphate(E4P)and phosphoenolpyruvate(PEP).Another pathway is the quinic acid pathway,which used quinic acid as substrate.On this basis,this study first detected the enzyme activities of the pathways related to the shikimic acid production process of Aspergillus cristatus and the expression of the corresponding genes,and studied the situation of the two pathways in Aspergillus cristatus;The aroF gene,the starting gene of shikimic acid pathway,was knocked out to study the effect of the enzyme activity of the shikimate pathway and the expression level of related genes,meanwhile,we also observed the phenotype difference of mutant strains and wild-type strains.The results of this study are as follows:1)The enzyme activity detection of shikimic acid pathway in Aspergillus cristatusThese results showed that aroF,quiB,aroD and aroE possessed activity under the sexual and asexual development stages,it indicated that shikimic acid pathway owned activity in Aspergillus cristatus.Meanwhile,these results showed that there are relationship between cleistothecium formation and aroF,quiB,aroD,and there are relationship between conida formation and aroE.2)Detection of the expression levels of genes in shikimic acid pathway from Aspergillus cristatusThese results showed that aroF,quiB,aroD and aroE were activitied under the sexual and asexual development stages in Aspergillus cristatus.The result showed aroF and quiB are related with the formation ascospore,aroE was related with the formation conidia.3)Observation on wild type and aroF mutant We compared the phenotype between wild-type and aroF mutant,the results showed that aroF mutant slow growth,it indicated that the gene is related with vegetative growth of Aspergillus cristatus.4)Detection enzyme activity and expression level of genes from wild-type and mutant strains of Aspergillus cristatusThe results showed that aroF positively regulate the enzyme activity and the expression level of quiB,aroD,and aroE genes in Aspergillus cristatusIn summary,the shikimic acid and quinic acid pathway own activity under the sexual and asexual development stages in Aspergillus cristatus.Meanwhile,aroF gene could regulate vegetative growth of Aspergillus cristatus.