LAUSR

As efficient reverse genetic tools are lacking, molecular genetics research has been limited in Hypsizygus marmoreus. In this study, we firstly constructed a gene‐silencing method using a dual promoter vector (DPV) which was driven by gpd and 35 S promoters. The DPV was introduced into H. marmoreus via a simple electroporation procedure and the highest silenced rate of ura3 gene was 76.6%, indicating that the DPV might be suitable for gene silencing in basidiomycete. In this silencing system, the endogenous orotidine 5′‐monophosphate decarboxylase gene (ura3) was used as a selectable marker. Besides, we also constructed another silencing system which could silence the ura3 and other genes (lcc1 encoded laccase1) together in H. marmoreus, and named it as co‐silencing system. In the co‐silenced transformants, we found that the mycelia were thinner and the growth was slower than in the wild‐type and control2 strains, which was accordant with the previous study of lcc1 gene, indicating that the selective efficiency of the RNAi‐mediated silencing of several genes might be increased by co‐silencing ura3. The development of this molecular tool might improve functional studies of multiple genes in the basidiomycete H. marmoreus and also provide a reference for studies of other basidiomycetes.