State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei Province, People's Republic of China.
PLoS One. 2012;7(12):e51853. doi: 10.1371/journal.pone.0051853. Epub 2012 Dec 14.
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 FINDINGS: We 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/SIGNIFICANCE: The 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.
蛹虫草是一种子囊菌毛毛虫真菌,由于其抗癌和免疫调节活性,已被用作传统中药多年。目前,这种有益真菌的人工培养已被广泛应用,可以满足市场需求,但在转录和翻译水平上对培养的蛹虫草发育阶段的系统分子研究尚未确定。
方法/主要发现:我们利用高通量 Illumina 测序获得了蛹虫草菌丝体和子实体的转录组。所有清洁读数都被映射到蛹虫草基因组上,大多数读数显示出完美的覆盖度。预测了选择性剪接和新转录本,以丰富数据库。基因表达分析表明,2113 个基因在菌丝体中上调,599 个在子实体中上调。进行了基因本体论 (GO) 和京都基因与基因组百科全书 (KEGG) 分析,以分析表达差异的基因。此外,研究了不同发育阶段中潜在的蛹虫草代谢差异。此外,通过一维凝胶电泳 (1-DGE) 与纳升电喷雾电离液相色谱串联质谱 (nESI-LC-MS/MS) 获得了菌丝体和子实体的蛋白质组数据。分别从菌丝体和子实体中检测到 359 和 214 种蛋白质。进一步进行了 GO、KEGG 和直系同源群 (COG) 分析,以更好地理解它们的差异。我们分析了这两种样品中一些值得注意的蛋白质的含量,包括凝集素、超氧化物歧化酶、糖苷水解酶和参与蛹虫草代谢的蛋白质,为进一步的蛋白质研究提供了重要信息。
结论/意义:通过转录组和蛋白质组分析,揭示了人工栽培蛹虫草菌丝体和子实体之间基因表达的差异。我们的研究为进一步开发和研究这种有前途的真菌提供了有效的资源。