Abstract
Gymnopilus junonius, a well-known poisonous mushroom, is distributed worldwide. It contains a hallucinogenic alkaloid psilocybin and several other bioactive compounds. The mitochondrial genome, a circular DNA molecule of 161,145 bp, comprises 15 protein-coding genes, 24 transfer RNA genes, and 2 ribosomal RNA genes. The guanine-cytosine content was 31.56%. Based on the mitochondrial genome sequence, a phylogenetic tree was constructed to demonstrate the phylogenetic relationship. In this study, the phylogenetic positions of G. junonius and its related genera were determined.
Keywords: Basidiomycota, Hymenogastraceae, phylogenetic analysis, poisonous mushroom
Gymnopilus junonius (Fr.) P.D. Orton is a well-known poisonous mushroom and contains psilocybin, a hallucinogenic alkaloid that is chemically related to the amino acid tryptophan (Chilton et al. 1979). Psilocybin is present in over 200 species of Basidiomycota mushrooms, including Gymnopilus (Guzmán et al. 1998). In addition, several bioactive compounds, such as gymnopilins, cerevisterol, acetylenic compounds, trichothecene, and tremulane sesquiterpenes, have been reported in Gymnopilus sp. (Kim et al. 2012; Lee et al. 2008, 2020). Hence, G. junonius is medicinally valuable and serves as a good source for research on hallucinogenic mushrooms. In this study, the complete mitochondrial genome of G. junonius was reported to confirm the phylogenetic relationship between the related genera.
Samples of G. junonius were collected from Gwangneung Forest, Pocheon-si, Gyeonggi-do, South Korea (37°45′N, 127°10′E) and deposited at the herbarium of the Korea National Arboretum (KH; voucher no. KA18-0872C). Genomic DNA was then isolated from the mycelium and subjected to the construction of an Illumina paired-end (PE) library according to the manufacturers’ protocol. The library was sequenced using Illumina PE sequencing at the PHYZEN Genomics Institute (Phyzen, Gyeonggi-do, South Korea). High-quality PE reads obtained after trimming were de novo assembled using the CLC genome assembler (v. 4.21, CLC Inc., Denmark). From the initially assembled contigs, those derived from the mitochondrial genome sequences were further processed to generate a single draft sequence, as described previously (Lee et al. 2018). The draft sequence was manually corrected and gap-filled using a series of PE read mapping. The complete mitochondrial genome sequence was annotated using GeSeq (https://chlorobox.mpimp-golm.mpg.de/geseq-app.html) and manually curated using the Artemis annotation tool (Rutherford et al. 2000) with NCBI BLASTN searches.
The complete mitochondrial genome sequence of G. junonius (GenBank Accession no. MW238478) was 161,145 bp in length, second largest among the previously reported mitogenomes of Agaricales. It consisted of 15 protein-coding genes, 24 transfer RNA genes, and 2 ribosomal RNA genes. The base composition was adenine (34.42%), cytosine (C) (15.41%), guanine (G) (16.14%), and thymine (34.01%). Overall GC content of the mitochondrial genome was 31.56%.
Phylogenetic analysis was performed based on the multiple alignments of protein-coding sequences in mitochondrial genomes (Kumar et al. 2016). Results revealed that Agrocybe aegerita (MF979820) was placed in a sister clade. In this study, the phylogenetic position of genus Gymnopilus was reported for the first time (Figure 1). Information on the mitochondrial genome sequence of Gymnopilus sp. would help understand species evolution and phylogenetic relationships between the related taxa.
Figure 1.
Phylogenetic tree of Gymnopilus junonius and 16 other related taxa. Sequences of protein-coding regions in the mitochondrial genome were aligned using MAFFT and MEGA 7.0. Numbers in the nodes indicate bootstrap support values (>80%) from 1000 replicates.
Acknowledgements
We would like to thank the researchers of Phyzen Co. for their assistance in software application and analysis.
Funding Statement
This work was supported by the Korea National Arboretum under Grant [No. KNA1-3-3, 20-3].
Disclosure statement
No potential conflict of interest was reported by the author(s).
Data availability statement
The genome sequence data that support the findings of this study are openly available in GenBank of NCBI at (https://www.ncbi.nlm.nih.gov/) under the accession no. MW238478. The associated BioProject, SRA, and Bio-Sample numbers are PRJNA700711, SRX10055256, and SAMN17838281, respectively.
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Associated Data
This section collects any data citations, data availability statements, or supplementary materials included in this article.
Data Availability Statement
The genome sequence data that support the findings of this study are openly available in GenBank of NCBI at (https://www.ncbi.nlm.nih.gov/) under the accession no. MW238478. The associated BioProject, SRA, and Bio-Sample numbers are PRJNA700711, SRX10055256, and SAMN17838281, respectively.