Characterization of a G protein α subunit encoded gene from the dimorphic fungus-Tremella fuciformis

Abstract

Tremella fuciformis is a dimorphic fungus which can undertake the reversible transition between yeast and pseudohypha forms. G protein α subunit (Gα) carries different signals to regulate a variety of biological processes in eukaryotes, including fungal dimorphism. In this study, a novel Gα subunit encoded gene, TrGpa1, was rstly cloned from T. fuciformis. The TrGpa1 open reading frame has 1059 nucleotides, and encodes a protein which belongs to the group I of Gα i superfamily. Furthermore, the role of TrGpa1 in the T. fuciformis dimorphism was analysed by gene overexpression and knockdown. Stable integration of the target gene into the genome was con rmed by PCR and Southern blot hybridization.Transformants with the highest and lowest TrGpa1 expression levels were selected via quantitative realtime PCR analysis and Western blot. Each transformant was compared with the wild-type strain about the morphological change under different environmental factors, including pH values, temperature, cultural time, inoculum size, and quorum-sensing molecules (farnesol and tyrosol). Comparing with the wild-type strain, the overexpression transformant always had higher ratios of pseudohyphae, while the knockdown transformant had less proportions of pseudohyphae. Therefore, the TrGpa1 involves in the dimorphism of T. fuciformis and plays a positive role in promoting pseudohyphal growth.

Transcriptome Analysis Reveals the Function of a G-Protein α Subunit Gene in the Growth and Development of Pleurotus eryngii

“…Transformed strains were obtained using ATMT according to the reported method [ 24 ]. Total RNA was isolated from the mycelia of the transformed strains, and the expression levels of PeGNAI were detected by qRT-PCR [ 25 ]. After dark culture at 25 ± 1 °C for 7 days, a fungal cake with a diameter of 9 mm at the edge of each colony was inoculated on PDA medium to obtain a sufficient quantity of P .…”

Study of Dimorphism Transition Mechanism of Tremella fuciformis Based on Comparative Proteomics

“…Isozyme electrophoresis showed differential expressions in esterase isozymes, polyphenol oxidase isozymes, and peroxidase isozymes produced by FBMds and DM [ 1 ]. TrGpa1 was also shown to be involved in the dimorphism of T. fuciformis and promote pseudohyphal growth [ 9 ]. There are few reports on the proteomics analysis of T. fuciformis dimorphism, and the dimorphism mechanism is still unknown.…”