Whole genome sequencing and genome annotation of the wild edible mushroom, Russula griseocarnosa.

Russula griseocarnosa is a species of edible ectomycorrhizal fungi with medicinal properties that grows in southern China. Total DNA was isolated from a fresh fruiting body of R. griseocarnosa and subjected to sequencing using Illumina Hiseq with the PacBio RS sequencing platform. Here, we present the 64.81 Mb draft genome map of R. griseocarnosa based on 471 scaffolds and 16,128 coding protein genes. The gene annotation of protein coding genes was used to obtain corresponding annotations by blastp. Phylogenetic analysis revealed a close evolutionary relationship of R. griseocarnosa to Heterobasidion irregulare and Stereum hirsutum in the core Russulales clade. The R. griseocarnosa genome encodes a repertoire of enzymes engaged in carbohydrate and polysaccharide metabolism, along with cytochrome P450s and secondary metabolite biosynthesis. The genome content of R. griseocarnosa provides insights into the genetic basis of its reported medicinal properties and serves as a reference for comparative genomics of fungi.

Optimization extraction and bioactivities of polysaccharide from wild Russula griseocarnosa.

The extraction conditions and biological activities of polysaccharides from wild Russula griseocarnosa (PRG) were investigated. Response Surface Methodology (RSM) with a Box-Behnken Design (BBD) was used to optimize extraction conditions. The optimal extraction parameters of PRG were as follows: extracting time 4 h, extraction temperature 77.3 °C and liquid-solid ratio 42.5 g/L. Furthermore, the data demonstrated that PRG exhibited antioxidant activities evidenced by reducing power to scavenge the DPPH, ABTS, hydroxyl radical and superoxide radical. PRG showed the activity of anti-cervical carcinoma cells Hela and Siha. In conclusion this study offered an efficient extraction method of wild Russula griseocarnosa polysaccharide, and the results suggested PRG had good antioxidant and inhibitory activities against cervical carcinoma cells, and PRG could be developed as a novel natural functional food.

Structural characterization of Russula griseocarnosa polysaccharide and its improvement on hematopoietic function.

The hematopoietic function of a polysaccharide derived from Russula griseocarnosa was demonstrated in K562 cells, and subsequently purified through chromatography to obtain RGP1. RGP1 is a galactan composed of 1,6-α-D-Galp as the main chain, with partial substitutions. A -CH3 substitution was detected at O-3 of 1,6-α-D-Galp. The possible branches at O-2 of 1,6-α-D-Galp was α-L-Fucp. In mice with cyclophosphamide (CTX)-induced hematopoietic dysfunction, RGP1 alleviated bone marrow damage and multinucleated giant cell infiltration of the spleen, increased the number of long-term hematopoietic stem cells, and regulated the levels of myeloid cells in the peripheral blood. Furthermore, RGP1 promoted the differentiation of activated T cells and CD4+ T cells without affecting natural killer cells and B cells. Proteomic analysis, detection of cytokines, and western blotting revealed that RGP1 could alleviate hematopoietic dysfunction by promoting the activation of CD4+ T cells and the Janus kinase/ signal transducer and activator of transcription 3 pathway. The present study provides experimental evidence to support the application of RGP1 in CTX-induced hematopoietic dysfunction.

Structural properties of glucan from Russula griseocarnosa and its immunomodulatory activities mediated via T cell differentiation.

The polysaccharide, RGP2, was isolated from Russula griseocarnosa and its immunostimulatory effects were confirmed in cyclophosphamide (CTX)-induced immunosuppressed mice. Following purification via chromatography, structural analysis revealed that RGP2 had a molecular weight of 11.82 kDa and consisted of glucose (Glc), galactose (Gal), mannose, glucuronic acid and glucosamine. Bond structure analysis and nuclear magnetic resonance characterization confirmed that the main chain of RGP2 was formed by →6)-ß-D-Glcp-(1→, →3)-ß-D-Glcp-(1→ and →6)-α-D-Galp-(1→, which was substituted at O-3 of →6)-ß-D-Glcp-(1→ by ß-D-Glcp-(1→. RGP2 was found to ameliorate pathological damage in the spleen and enhance immune cell activity in immunosuppressed mice. Based on combined multiomics analysis, RGP2 altered the abundance of immune-related microbiota (such as Lactobacillus, Faecalibacterium, and Bacteroides) in the gut and metabolites (uridine, leucine, and tryptophan) in the serum. Compared with immunosuppressed mice, RGP2 also restored the function of antigen-presenting cells, promoted the polarization of macrophages into the M1 phenotype, positively affected the differentiation of helper T cells, and inhibited regulatory T cell differentiation through the protein kinase B (AKT)/mechanistic target of rapamycin (mTOR) pathway, ultimately exerting an immune boosting function. Overall, our findings highlight therapeutic strategies to alleviate CTX-induced immunosuppression in a clinical setting.

Whole genome sequencing of an edible and medicinal mushroom, Russula griseocarnosa, and its association with mycorrhizal characteristics.

Russula griseocarnosa is a well-known ectomycorrhizal mushroom, which is mainly distributed in the Southern China. Although several scholars have attempted to isolate and cultivate fungal strains, no accurate method for culture of artificial fruiting bodies has been presented owing to difficulties associated with mycelium growth on artificial media. Herein, we sequenced R. griseocarnosa genome using the second- and third-generation sequencing technologies, followed by de novo assembly of high-throughput sequencing reads, and GeneMark-ES, BLAST, CAZy, and other databases were utilized for functional gene annotation. We also constructed a phylogenetic tree using different species of fungi, and also conducted comparative genomics analysis of R. griseocarnosa against its four representative species. In addition, we evaluated the accuracy of one already sequenced genome of R. griseocarnosa based on the internal transcribed spacer (ITS) sequencing of that type of species. The assembly process resulted in identification of 230 scaffolds with a total genome size of 50.67 Mbp. The gene prediction showed that R. griseocarnosa genome included 14,229 coding sequences (CDs). In addition, 470 RNAs were predicted with 155 transfer RNAs (tRNAs), 49 ribosomal RNAs (rRNAs), 41 small noncoding RNAs (sRNAs), 42 small nuclear RNAs (snRNAs), and 183 microRNAs (miRNAs). The predicted protein sequences of R. griseocarnosa were analyzed to indicate the existence of carbohydrate-active enzymes (CAZymes), and the results revealed that 153 genes encoded CAZymes, which were distributed in 58 CAZyme families. These enzymes included 78 glycoside hydrolases (GHs), 34 glycosyl transferases (GTs), 30 auxiliary activities (AAs), 2 carbohydrate esterases (CEs), 8 carbohydrate-binding modules (CBMs), and only one polysaccharide lyase (PL). Compared with other fungi, R. griseocarnosa had fewer CAZymes, and the number and distribution of CAZymes were similar to other mycorrhizal fungi, such as Tricholoma matsutake and Suillus luteus. Well-defined effector proteins that were associated with mycorrhiza-induced small-secreted proteins (MiSSPs) were not found in R. griseocarnosa, which indicated that there may be some special effector proteins to interact with host plants in R. griseocarnosa. The genome of R. griseocarnosa may provide new insights into the energy metabolism of ectomycorrhizal (ECM) fungi, a reference to study ecosystem and evolutionary diversification of R. griseocarnosa, as well as promoting the study of artificial domestication.

Bacterial Community Selection of Russula griseocarnosa Mycosphere Soil.

Russula griseocarnosa is a wild, ectomycorrhizal, edible, and medicinal fungus with high economic value in southern China. R. griseocarnosa fruiting bodies cannot be artificially cultivated. To better understand the effects of abiotic and biotic factors on R. griseocarnosa growth, the physicochemical properties of R. griseocarnosa and its associated bacterial communities were investigated in two soil types (mycosphere and bulk soil) from Fujian, Guangdong, and Guangxi Provinces. The results revealed that the diversity, community structure, and functional characteristics of the dominant mycosphere bacteria in all geographical locations were similar. Soil pH and available nitrogen (AN) are the major factors influencing the mycosphere-soil bacterial communities' structure. The diversity of soil bacteria is decreased in R. griseocarnosa mycosphere when compared with the bulk soil. Burkholderia-Paraburkholderia, Mycobacterium, Roseiarcus, Sorangium, Acidobacterium, and Singulisphaera may also be mycorrhiza helper bacteria (MHB) of R. griseocarnosa. The functional traits related to the two-component system, bacterial secretion system, tyrosine metabolism, biosynthesis of unsaturated fatty acids, and metabolism of cofactors and vitamins were more abundant in R. griseocarnosa mycosphere soil. The mycosphere soil bacteria of R. griseocarnosa play a key role in R. griseocarnosa growth. Application of management strategies, such as N fertilizer and microbial fertilizer containing MHB, may promote the conservation, propagation promotion, and sustainable utilization of R. griseocarnosa.

The complete mitochondrial genome of a wild edible mushroom, Russula griseocarnosa.

Russula griseocarnosa is a wild edible ectomycorrhizal mushroom in southern China. In this study, we assembled the complete mitochondrial genome of R. griseocarnosa. Its total length was 60995 bp with a GC content of 21% and contained a total of 52 genes, including 14 standard protein-coding genes, two rRNA genes, 21 tRNA genes and 15 free-standing open reading frames (ORFs). Phylogenetic analysis reflected that the evolutionary processes between R. griseocarnosa and some agaricomycetes.

Data on the genome analysis of the wild edible mushroom, Russula griseocarnosa.

In the present article, we report data on the whole genome sequence of a wild edible and medicinal ectomycorrhizal fungus Russula griseocarnosa. The R. griseocarnosa genome consists of 64.81 Mb with a GC-pair content of 49.41%. The genome assembly consists of 471 scaffolds and 16128 coding protein genes. The coding protein genes was annotated in different databases (GO, KEGG and CAZys), respectively. The whole genome sequence and functional annotation provide important information for ectomycorrhizal fungus, which can be used as a basis for cultivation and breeding of R. griseocarnosa. The Whole Genome project of Russula griseocarnosa has been deposited at DDBJ/ENA/GenBank under the accession RMVF00000000. The version described is RMVF01000000. To further interpretation of the data provided in this article, please refer to the research article 'Whole genome sequencing and genome annotation of the wild edible mushroom, Russula griseocarnosa' [1].

Purification, characterization and anti-tumor activities of polysaccharides extracted from wild Russula griseocarnosa.

The anti-tumor activity of a novel polysaccharide, PRG1-1, obtained from Russula griseocarnosa sporocarp was investigated in this paper. PRG1-1 has a molecular weight of 630kDa and was extracted and purified using DEAE-cellulose and gel filtration chromatography from crude polysaccharide extract of R. griseocarnosa sporocarp. PRG1-1 was composed of glucose, galactose, mannose, xylose and fructose, in a molar ratio of 66.5:29.2:3.17: 0.663:0.447, respectively. Purified PRG1-1 significantly reduced cell viability, increased the production of lactate dehydrogenase (LDH) and reactive oxygen species (ROS), and enhanced the apoptotic rate in HeLa and SiHa cells. Furthermore, after 24h of PRG1-1 exposure the expression levels of cleaved PARP and caspase-3 were increased and mitochondrial cytochrome c was induced to release to the cytosol. Collectively, our results suggested that the cytotoxicity effects of PRG1-1 on human cervical carcinoma are associated with the apoptotic pathway. These data indicate the promising potential of bioactive PRG1-1 as natural agent to inhibit tumor cell proliferation in the treatment of cervical carcinoma.