Study On The Functions Of CYP51s(14?-demethylase Gene) In Arthrobotrys Oligospora

CYP51 is a type of P450 enzymes in the biosynthesis of steroids.It has been regarded as an important target of antifungal drugs for the treatment of Human African Trypanosomiasis.Bioinformatic analysis revealed three CYP51 genes in A.oligospora genome including AOL_s00076g231,AOL_s00080g314 and AOL_s00078g599.Three CYP51 mutant strains were obtained through fungal genetics engineering and evaluated for their morphology and viability including mycelial growth.sporulation,spore germination,trap formations and nematode predating ability.Compared with the wild-type strain,three mutants all displayed significantly increased spore germination while the mutant A A OL_s00080g314 showed significantly decreased sporulation and the trap formations.HPLC and GC-MS analysis revealed that the extracts of the fermentation broth of mutant ?AOL_s00080g314 yielded 10 unique compounds,including geranylgeraniol,that had been reported as to show antiviral and antitumor activity.Mutant AOL_s00076g231 yielded 16 unique compounds and mutant AOL_s00080g314 produced 9 unique compounds.RNA-seq analysis revealed that the knockout of the gene AOL_s00076g231 caused the up-regulated transcription of 1145 genes and the down-regulated transcription of 246 genes.The disruption of the gene AOL_s00078g599 led to the up-regulated transcription of 123 genes and down regulated transcription of 36 genes.Among them,16 genes in up-regulated transcription and 24 genes in down-regulated transcription were shared.The highest down-regulated transcription gene of two mutants was elucidated to be AOL_s00080g150,which encoded a UV-damage endonuclease.Compared with the wild-type strain,the highest up-regulated transcription gene of AAOL_s00076g231 is AOL_s00006g314,which encoded a hypothetical protein.AAOL_s00078g599 is AOL₀0043g217,which encoded a phospholipase A2.GO enrichment analysis revealed that the disruption of AOL_s00076g231 and AOL_s00078g599 displayed some similar effect on the differential transcription of genes involved in cell composition,metabolic process and catalytic activity.KEGG pathway enrichment analysis suggested that differentially expressed genes should be mainly involved in carbohydrate metabolism,amino acid metabolism and secondary metabolite synthesis pathways.It seemed that A OL_s00076g231 and AOL_s00078g599 might play negative regulatory roles in the growth and development of A.oligospora.