Genome-Wide Transcriptome and Proteome Analysis on Different Developmental Stages of Cordyceps militaris

Abstract

Background Cordyceps militaris, an ascomycete caterpillar fungus, has been used as a traditional Chinese medicine for many years owing to its anticancer and immunomodulatory activities. Currently, artificial culturing of this beneficial fungus has been widely used and can meet the market, but systematic molecular studies on the developmental stages of cultured C. militaris at transcriptional and translational levels have not been determined.Methodology/Principal FindingsWe utilized high-throughput Illumina sequencing to obtain the transcriptomes of C. militaris mycelium and fruiting body. All clean reads were mapped to C. militaris genome and most of the reads showed perfect coverage. Alternative splicing and novel transcripts were predicted to enrich the database. Gene expression analysis revealed that 2,113 genes were up-regulated in mycelium and 599 in fruiting body. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were performed to analyze the genes with expression differences. Moreover, the putative cordycepin metabolism difference between different developmental stages was studied. In addition, the proteome data of mycelium and fruiting body were obtained by one-dimensional gel electrophoresis (1-DGE) coupled with nano-electrospray ionization liquid chromatography tandem mass spectrometry (nESI-LC-MS/MS). 359 and 214 proteins were detected from mycelium and fruiting body respectively. GO, KEGG and Cluster of Orthologous Groups (COG) analysis were further conducted to better understand their difference. We analyzed the amounts of some noteworthy proteins in these two samples including lectin, superoxide dismutase, glycoside hydrolase and proteins involved in cordycepin metabolism, providing important information for further protein studies.Conclusions/SignificanceThe results reveal the difference in gene expression between the mycelium and fruiting body of artificially cultivated C. militaris by transcriptome and proteome analysis. Our study provides an effective resource for the further developmental and medicinal research of this promising fungus.

Entomotoxic and nematotoxic lectins and protease inhibitors from fungal fruiting bodies

“…These fungi are characterized by the formation of dikaryotic hyphae and include the phyla Ascomycota and Basidiomycota. Fruiting body formation is independent of the lifestyle (saprotrophic or biotrophic) of these fungi and sometimes coupled to the preceding formation of sclerotia-compact masses of hardened mycelium containing food reserves (Martin et al 2008;Stajich et al 2010;Teichert et al 2014;Yin et al 2012). Since the fruiting bodies produce and disperse the sexual spores, defense of these structures against fungivores, including predators, grazers, and parasites, is essential for fungal reproduction.…”

Comparative mitochondrial genomics toward exploring molecular markers in the medicinal fungus Cordyceps militaris

“…Biologically active compounds (e.g., cordycepin, polysaccharides, cordymin) isolated from the fungus exhibit a variety of pharmacological effects, including anti-cancer, antioxidant, anti-inflammatory, immune-enhancing, or antifungal activities2345. In addition to the interest in artificial cultivation and pharmacological effects, researchers have studied the fungus broadly from the viewpoints of genomics6, transcriptomics78, methylome9, and proteomics7.…”

Simple Genetic Tools to Study Fruiting Body Development in Fungi

“…Fruiting body mutants are now available for many fungi other than P. anserina, including Sordaria macrospora [27], N. crassa [28], A. nidulans [29] and Coprinopsis cinerea (formely Coprinus cinereus) [30,31] and could be analyzed using similar methods. They are complementary to the now more widespread large scale analyses of transcriptomes during fruiting body development as performed in the ascomycetes Fusarium graminearum and Fusarium verticilloides [32][33][34], N. crassa [35,36], S. macrospora [37], Pyronema confluens [38], Tuber melanosporum [39][40][41], Cordyceps militaris [42] and Ophiocordyceps sinensis [43], as well as in the basidiomycetes C. cinerea [44], Moniliophthora perniciosa [45], Agrocybe aegerita [46], Lentinula edodes [47,48], Auricularia polytricha [49] and Ganoderma lucidum [50].…”