The Analysis And Study Of Genome,transcriptome And Interaction With Hosts Of Epidermophyton Floccosum

Dermatophytes cause the fourth most common infections,and easily to recur in clinic.Epidermophyton floccosum is one of the most common dermatophytes,which can cause infections throughout the world especially in India and Iran,skin and nails other than hairs are the common affected sites.E.floccosum belongs to the genus Epidermophyton and it's the only species,which means that it possesses unique traits.Lacking of animal model and relative modern research tools for E.floccosum,there are still many unknown aspects of it.Thus,we perform a series of experiments from basic traits,molecular,genome to transcriptome to reveal the reason behind its uniqueness and the interaction mechanism with human host.In the first part,we analyzed the morphology and enzymatic activities in vitro of 19 E.floccosum strains.It revealed that E.floccosum have four main morphology types on PDA medium,and the clinical and ecological traits might have some relations with keratinase and lipase activities of it.Then we used two molecular methods including multi-locus and AFLP to divide E.floccosum strains into four main gene types,but we failed to find the correlation with morphological,enzymatic and geographical differences.High quality whole genome sequences of E.floccosum was obtained by combining next generation with single molecular sequencing methods in the second part of this paper.And we found that E.floccosum owns a complete system of energy and nucleic metabolism to adapt itself to changing environments.By comparing its genome sequence with other dermatophytes,we found that E.floccosum has the largest genome size and different directions in some locus,and Tf2 and Tcl-mariner transposons are considered to be the reason.We further revealed some aspects concerning the pathogenic and ecological features of E.floccosum as followings.1)The number and type of adhesion factors especially for 1,3-beta-glucanosyltransferase gel3.2)The number of Cdo1,sub1,sub3,sub7 for keratin hydrolyzing.3)The number of CEs and CBMs in CAzy family.4)The type and number of LysM domain.4)Serine/threonine domains.Furthermore,the reproduction way of E.floccosum was asexual and the mating type was MAT 1-2,and confirmed in both molecular and genomic levels.Finally,for the purpose of revealing key regulatory genes with proteolysis and host interaction of E.floccosum to laying foundation for biomarkers and target therapy in clinic,RNA-seq and dual RNA-seq methods were adopted to analyze gene expressions of E.floccosum in vitro and co-culture with human cells separately.E.floccosum showed special regulatory mechanism for proteolysis compared with Nannizzia gypseum which has the most relationship with it.For E.floccosum itself,metalloproteases and DPPV played important roles in the process of keratin hydrolysis.FAD,MFS transporter,phospholipase D were important virulence factors for infection process.The expression of KRT75 gene might related to rare hair infections,and CDC7 protein kinase was thought to be one of the key regulatory factors for asexual reproduction.For cells part,the up-regulation gene expressions related to cell proliferation and differentiation,cytoskeleton,extracellular matrix remodeling were protection strategies against inj uries from E.floccosum invasion.