Construction and application of a gene silencing system using a dual promoter silencing vector in Hypsizygus marmoreus

Abstract

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.

Comparative transcriptome analysis on candidate genes associated with fruiting body growth and development in Lyophyllum decastes

“…For example, Volvariella volvacea produces multiple forms of extracellular laccase when grown in specific culture conditions, and some laccase genes may play an important role in the development of fruiting bodies ( Chen et al, 2003 ; Lu et al, 2015 ). Studies have found that amplifying the operation of laccase stimulates the initiation of H. marmoreus primordia and elevates the production of fruiting bodies ( Zhang et al, 2015b ; Zhang et al, 2016 ). Organisms’ carbohydrate metabolismrely heavily on lignocellulolytic enzymes, which are crucial for the transformation of the lignocellulolytic matrix during fungal growth and serve as a necessary physiological function ( Elisashvili et al, 2002 ; Bánfi et al, 2015 ).…”

Optimization, Characterization and Antioxidant Activities of Selenized Polysaccharides from Hypsizygus marmoreus

“…Polysaccharide is one of the main components of H. marmoreus. Studies on PHM mainly focused on biological activities, such as antitumor (Yasukawa et al, 1994), immune enhancer (Zhang et al, 2015a), antivirus (Song et al, 2013), antioxidation (Liu et al, 2018;Huang et al, 2015) and antibacterial (Xing et al, 2007) properties through molecular structural characteristic (Lingyun, 2014), extraction (Zhang et al, 2017a;2017b;Harada et al, 2005) and component analyses (Tao et al, 2007) to date. However, the study on the chelation of PHM and microelement has not been reported in the literature.…”