Project description:Phellinus linteus, also known as the sanghuang mushroom, is a medicinal mushroom that has been recognized as beneficial to health for more thousands of years. Among its diverse valuable secondary metabolites, the yellow-brown styrylpyrone pigment hispidin has garnered significant attention due to its various pharmacological effects. However, recently after detailed morphological and molecular phylogenetic studies, the correct scientific name of the true sanghuang strains was shown not to be P. linteus but Sanghuangporus sanghuang. As the incorrect binomial name P. linteus has long been misleadingly referred, there is a need to evaluate the safety of S. sanghuang. Moreover, the growing conditions can impact the secondary metabolite profile of the fungi. Hence, this study is the first to optimize hispidin production and to investigate the genotoxic and oral toxic effects of hispidin-enriched S. sanghuang mycelia. In order to induce the biosynthesis of hispidin, 15 different culture media consisting of five carbon sources, five nitrogen sources, and five initial pH conditions were screened. Glucose and yeast extract at an initial pH of 5 were found to be the most suitable carbon and nitrogen sources, respectively, for the optimal growth and production of hispidin. Moreover, the production of hispidin was 3 mg/g in a 20-ton bioreactor under optimal conditions. Furthermore, the ames test, in vitro chromosome aberration test, acute oral toxicity test, and bone marrow micronucleus test were used to detect toxicological properties of 3 mg/g hispidin-enriched S. sanghuang mycelia. In all tests, there was no statistically significant difference between the mycelia and the negative control. Based on the results obtained, the present study demonstrates that 3 mg/g hispidin-enriched S. sanghuang mycelia has a very low order of toxicity, which supports its safety for human consumption.
Project description:BackgroundThe microbial symbionts of macrofungal fruiting body have been shown to play momentous roles in host growth, development, and secondary metabolism. Nevertheless, there is no report on the fungal diversity of Sanghuangporus, a medicinal and edible homologous macrofungus as "forest gold", which has good effects on antioxidation, boosting immunity and curing stomachache. Here, the diversity and functional group of fungi associated with the fruiting body of the most widely applied S. vaninii were characterized by high-throughput sequencing and FUNGuild tool for the first time.ResultsTotal 11 phyla, 34 classes, 84 orders, 186 families, and 328 genera were identified in the fruiting body, and our results revealed that the fungal community was dominated by the host fungal taxonomy with absolute superiority (more than 70%), namely, Basidiomycota, Agaricomycetes, Hymenochaetales, Hymenochaetaceae, and genus of Phellinus corrected to Sanghuangporus. Simultaneously, the reads allocated into non-host fungal operational taxonomic units were largely dominated by Ascomycota, Sordariomycetes, Sordariales, Mortierellaceae, and Mortierella. Furthermore, the endophytic fungi were assigned into three trophic modes of "saprotroph" (53.2%), "symbiotroph" (32.2%), and "pathotroph" (14.1%), in which the category of "plant pathogen" was highest enriched with relative abundance of 91.8%, indicating that the endophytic fungi may have the potential to adjust the growth and metabolism of host S. vaninii.ConclusionAltogether, this report firstly provided new findings that can be inspiring for further in-depth studies to exploit bioactive microbial resources for increased production of Sanghuangporus via coculture, as well as to explore the relationship between macrofungi and their associated endophytes.
Project description:miRNA-like RNAs (milRNAs) have been recognized as sequence-specific regulators of posttranscriptional regulation of gene expression in eukaryotes. However, the functions of hundreds of fungal milRNAs in the biosynthesis of metabolic components are obscure. Sanghuangporus produces diverse bioactive compounds and is widely used in Asian countries. Here, genes encoding two Dicers, four Argonautes, and four RdRPs were identified and characterized in Sanghuangporus vanini. Due to the lack of an efficient gene manipulation system, the efficacy of spray-induced gene silencing (SIGS) was determined in S. vanini, which showed efficient double-stranded RNA (dsRNA) uptake and gene silencing efficiency. SIGS-mediated gene knockdown showed that SVRDRP-3, SVRDRP-4, SVDICER-1, and SVDICER-2 were critical for mycelial biomass, flavonoid, triterpenoid, and polysaccharide production. Illumina deep sequencing was performed to characterize the milRNAs from S. vanini mycelium and fruiting body. A total of 31 milRNAs were identified, out of which, SvmilR10, SvmilR17, and SvmilR33 were Svrdrp-4- and Svdicer-1-dependent milRNAs. Importantly, SIGS-mediated overexpression of SvmilR10 and SvmilR33 resulted in significant changes in the yields of flavonoids, triterpenoids, and polysaccharides. Further analysis showed that these milRNA target genes encoding the retrotransposon-derived protein PEG1 and histone-lysine N-methyltransferase were potentially downregulated in the milRNA overexpressing strain. Our results revealed that S. vanini has high external dsRNA and small RNA uptake efficiency and that milRNAs may play crucial regulatory roles in the biosynthesis of bioactive compounds. IMPORTANCE Fungi can take up environmental RNA that can silence fungal genes with RNA interference, which prompts the development of SIGS. Efficient dsRNA and milRNA uptake in S. vanini, successful dsRNA-targeted gene block, and the increase in intracellular miRNA abundance showed that SIGS technology is an effective and powerful tool for the functional dissection of fungal genes and millRNAs. We found that the RdRP, Dicer, and Argonaute genes are critical for mycelial biomass and bioactive compound production. Our study also demonstrated that overexpressed SVRDRP-4- and SVDICER-1-dependent milRNAs (SvmilR10 and SvmilR33) led to significant changes in the yields of the three active compounds. This study not only provides the first report on SIGS-based gene and milRNA function exploration, but also provides a theoretical platform for exploration of the functions of milRNAs involved in biosynthesis of metabolic compounds in fungi.
Project description:BackgroundThe Inonotus obliquus mushroom, a wondrous fungus boasting edible and medicinal qualities, has been widely used as a folk medicine and shown to have many potential pharmacological secondary metabolites. The purpose of this study was to supply a global landscape of genome-based integrated omic analysis of the fungus under lab-growth conditions.ResultsThis study presented a genome with high accuracy and completeness using the Pacbio Sequel II third-generation sequencing method. The de novo assembled fungal genome was 36.13 Mb, and contained 8352 predicted protein-coding genes, of which 365 carbohydrate-active enzyme (CAZyme)-coding genes and 19 biosynthetic gene clusters (BCGs) for secondary metabolites were identified. Comparative transcriptomic and proteomic analysis revealed a global view of differential metabolic change between seed and fermentation culture, and demonstrated positive correlations between transcription and expression levels of 157 differentially expressed genes involved in the metabolism of amino acids, fatty acids, secondary metabolites, antioxidant and immune responses. Facilitated by the widely targeted metabolomic approach, a total of 307 secondary substances were identified and quantified, with a significant increase in the production of antioxidant polyphenols.ConclusionThis study provided the comprehensive analysis of the fungus Inonotus obliquus, and supplied fundamental information for further screening of promising target metabolites and exploring the link between the genome and metabolites.
Project description:To evaluate the novel strategy of oleic acid and fungal elicitor (made from Aspergillus niger) to elicit betulinic acid biosynthesis in medicinal mushroom Inonotus obliquus, we conduct the stimulatory effects investigation for synthesizing betulinic acid from betulin. HPLC results indicated oleic acid and fungal elicitor were effective stimulators. The supplementation of 1.0 g/L oleic acid led to the highest increase of betulinic acid either in dry mycelia or fermentation broth by 2-fold of the control. Fungal elicitor at 45 mg/L markedly increases mycelia growth by 146.0% and enhance intracellular betulinic acid accumulation by 429.5% as compared to the controls. Quantification of transcription levels determined that oleic acid, fungal elicitor and their combinations could induce the expressions of key genes involved in betulinic acid biosynthesis, such as HMG-CoA reductase and squalene synthase. These findings indicated that oleic acid and fungal elicitor could enhance betulinic acid metabolism by up-regulating key genes expression.
Project description:Selenium (Se) is an essential nutrient element in human physiological metabolism and immune function. Supplementation of bioavailable Se will confer benefit on human life, especially when intake of this nutrient is inadequate. The edible and medicinal mushroom Antrodia camphorata is a unique fungus endemic to Taiwan, which has shown high therapeutic and nutritive value. This study is the first to demonstrate that A. camphorata can assimilate and transform sodium selenite into organic selenium. With an initial concentration of Se (IV) at 10 mg/L in 100 mL of the medium at 25 °C, the total selenium content in Se-enriched A. camphorata mycelia was 1281.3 ± 79.2 µg/g, in which the organic selenium content accounted for 88.1%. Further analysis demonstrated that selenium-enriched polysaccharide was the main form of Se present in A. camphorata (61.5% of the organic selenium). Four water-soluble Se-polysaccharide fractions were separated from A. camphorata, and ACP II was the major fraction of Se-polysaccharide. The scavenging efficiency of Se-polysaccharides on DPPH and ABTS radicals was determined, proving that selenium enrichment dramatically improved the in vitro antioxidant capacity of A. camphorata polysaccharide. Therefore, the selenium accumulation and transformation ability of A. camphorata provides an opportunity for developing this beneficent fungus into a novel selenium-enriched dietary or medicinal supplement.
Project description:In this study, the complete mitochondrial genome of Sanghuangporus vaninii Zhehuang-1 was determined. Genomic DNA samples individually collected from a population in southeastern China (Qiandaohu, Zhejiang Province, N29°44'24'', E118°52'48'') were sequenced with an Illumina NovaSeq 6000. The complete mitochondrial genome was 97,345 bp in length and consisted of 22 protein-coding genes (PCGs), 29 tRNAs, 3 ribosomal RNAs, and a control region. Sanghuangporus vaninii has a mitochondrial gene arrangement that is similar to that of S. sanghuang. Phylogenetic analysis performed using ML methods based on the complete mitogenome sequence showed that S. vaninii is a member of the Polyporaceae. The complete mitogenome sequence provides important data for further study of the Inonotus linteus complex.