Screening And Preliminary Function Analysis Of Differentially Expressed Genes At Three Development Stages Of Xerocomus Spadiceus

With high economic value,Xerocomus spadiceus is widely distributed in Yunnan province and deeply loved by people.It belongs to Xerocomus of Boletaceae of Boletales of Agaricomycetes of Basidiomycota of Fungi.It not only tastes delicious,but also has anti-oxidant,anti-tumor and other medicinal effects.Boletes is a great of ectomycorrhizal fungi that are symbiotic with plants,and the mycelia induced under pure culture conditions indoors grew slowly,would not kink the primordia,which has been the crux of artificial culture.In our laboratory small primordia of X.spadiceus have been induced,therefore,in the paper the species was used as research material,with combing previous transcriptome sequencing data,the difference genes on three developmenta stages of mycelia,primordia and fruit bodies were analyzed.32 differential genes were optimally selected by multiple comparisons of single and composite samples.On this basis,three differentially expressed genes of up-regulation were finally chosed,their expression quantities in mycelia and primordia cultivated on8 th days and 15 th days were detected respectively using RT-qPCR technology,and the role of three genes in the hyphae growth,primordia and fruit bodies development were predicted with combining their database function annotation results.These above has laid a foundation for further determination candidate genes that participate in the formation of primordia and fruit bodies,and for the study on the correlative regulation genes and their functions of fruit bodies development.The main results were as follows:1.From differentially expressed gene sets of fruit bodies-vs-primordia,the selected c20382.graph-c1 gene(named xs-lectin)was up-regulated,and annotated as a fungal agglutinin gene by Pfam database.The gapdh gene used as the internal reference.RT-qPCR analysis showed that relative expression amount of xs-lectin gene in primordia cultured on 8th days and 15 th days were 675.6 and 137.2 times than that of the fruit bodies respectively.It was speculated that the gene was involved in the morphogenesis of the primordia.2.The selected c18954.graph-c1 gene(named xs-hyd 1)was both up-regulated in the differentially expressed gene sets of fruitbodies-vs-mycelia and fruit bodies-vs-primordia,and annotated as the hydrophobin gene by Pfam database.The gapdh gene used as an internal reference,RT-qPCR analysis showed that the relative expression levels of xs-hyd 1 gene in mycelia cultured on 8th days and 15 th days were207.9 and 26 times than that of fruit bodies respectively,which in primordia on 8th days and 15 th days were 337.8 and 6.5 times than that of fruit bodies respectively,the expression of mycelia and primordia was both relative higher on 8th days.It was presumed that the gene expression was activated during hypha inducted,growth and primordia morphogenesis.3.The selected c22444.graph-c7 gene(named xs-hyd 2)was up-regulated in the differentially expressed gene sets of fruit bodies-vs-mycelia,annotated as hydrophobin gene by Pfam database.To RT-qPCR analysis results,the relative expression amount of xs-hyd 2 gene in mycelia cultured on 8th days and 15 th days was 5404.7 and 548.7times than that of fruit bodies respectively.The gene was highly expressed during the induction and growth stages of mycelia,and it was speculated that it was related to the growth of mycelia.