Characterization of the role of glycogen synthase kinase in fruiting body development in model mushroom Coprinopsis cinerea through transcriptome profiling and molecular editing

Ph.D. ; Coprinopsis cinerea is a model mushroom to study developmental processes in homobasidiomycetous fungi. The development of C. cinerea depends on the sensing of environmental conditions, such as light, temperature, humidity and nutrients. The signal transduction pathway from the environmental condition sensors to the regulators and reproduction has been an important research topic as the knowledge of that is still fragmentary. As protein kinases not only are crucial to the signal transduction, but also can be master switches of development. The functions of glycogen synthase kinase (GSK3) have been well-studied in animal, plant and yeast. However, information on its roles in basidiomycetous fungi is still limited. ; By RNA interference, 74% of stage 1 primordium samples retarded the fruiting body development by transient knockdown with Cc.Gsk3 siRNA. The efficiency of knockdown by siRNA showed by reducing the expression of Cc.Gsk3 gene significantly as well as eIF1 gene. Besides, application of GSK3 inhibitor, like lithium chloride (LiCl) or CHIR99021 trihydrochloride, induced enhanced mycelial growth and inhibited fruiting body formation in C. cinerea. RNA-Seq of LiCl-treated C. cinerea resulted in a total of 14128 unigenes. There were 1210 differentially expressed genes (DEGs) between the LiCl-treated samples and control samples in the mycelium stage (first time point), whereas 1402 DEGs were detected at the stage when the control samples formed hyphal knots and the treatment samples were still in mycelium (second time point). Downregulated protein kinase genes in LiCl treated samples were involved in metabolism pathways and cellular processes and signaling pathway, indicated that the inhibited GSK3 may suppress those kinases expression and further arrested fruiting body formation. This study suggests that GSK3 activity is essential for fruiting body development. ; I developed a system of CRISPR/ Cas9 RNP with double strain template donor, which contains selective mark hygromycin cassette. CcPka1 gene ...