Geographical traceability and multielement analysis of edible and medicinal fungi: Taking Wolfiporia cocos (F.A. Wolf) Ryvarden and Gilb. as an example.

Different geographical environment has a certain influence on the accumulation of fungi elements and chemical components. However, our knowledge is limited to elucidate the fungi elements in response to heterogeneous environmental and the quality differences among different habitats. Here, multielement analysis, FTIR spectrum, and feature-level fusion technique combined with chemometrics were used to study Wolfiporia cocos from different geographical areas, different sampling sites and different altitude sources. From the results, (1) there is significant difference in element content of samples from different sampling sites and no positive correlation with geographical ranges. (2) There is a correlation between elevation and elements, and relatively low elevation (<1,800 m) is conducive to the enrichment of elements. (3) From the perspective of elements, the W. cocos in Yuxi have relatively better quality. (4) FTIR and feature-level models can well realize origin identification. The SVM models are better than the PLS-DA models, and the feature-level model is better than the single FTIR models. In summary, this study demonstrated that the developed method was reliable and could realize the genuineness evaluation and origin identification of W. cocos. The results have implications for the establishment of the technology system of geographical traceability and the development of high-quality geographical indication products of W. cocos.

Comparative Studies on Polysaccharides, Triterpenoids, and Essential Oil from Fermented Mycelia and Cultivated Sclerotium of a Medicinal and Edible Mushroom, Poria Cocos.

Poria cocos, an important medicinal and edible fungus, is well known in East Asia. The main active components are water-soluble polysaccharides (WPS) and triterpenoids. Due to the growing market demand, long cultivation period, and consumption of pine trunk during cultivation, alternative methods for producing P. cocos or its active components should be investigated. In this study, WPS, triterpenoids, monosaccharide composition, and essential oil in fermented mycelia and cultivated sclerotium were analyzed using UV spectrophotometry, HPLC, pre-column derivatization, and HS-GC/MS, respectively. Our results showed that the WPS and triterpenoids in mycelia are several times higher than those in sclerotium. Among the 62 compounds identified by HS-GC/MS analysis from the essential oil obtained from the fermentation media and a fresh external layer, the two main fragrances in common were linalool and methyl phenylacetate. Our results suggested that it is applicable to produce polysaccharides and triterpenoids by the fermentation of P. cocos, and a strategy to improve triterpenoid production in the fermentation process was proposed.

Differentiation and comparison of Wolfiporia cocos raw materials based on multi-spectral information fusion and chemometric methods.

In order to achieve the target of deeper insight into the differentiation and comparison of Wolfiporia cocos, a total of 350 samples including distinct growth patterns, various collection regions and different medicinal parts were investigated using multi-spectral information fusion based on ultraviolet (UV) and Fourier transform infrared (FT-IR) spectroscopies coupled with chemometrics. From the results, the discrimination of samples was obtained successfully and good classification performances were shown according to partial least squares discriminant analysis (PLS-DA) models. Comparatively, the distinctness of chemical information in the two medicinal parts of W. cocos were much more than that in the same part with different growth patterns and collection areas. Meanwhile, an interesting finding suggested that growth patterns rather than geographical origins could be the dominant factor to effect the chemical properties of the same part samples, especially for the epidermis. Compared with the epidermis samples, there were better quality consistency for the inner part of W. cocos. Totally, this study demonstrated that the developed method proved to be reliable to perform comparative analysis of W. cocos. Moreover, it could provide more comprehensive chemical evidence for the critical supplement of quality assessment on the raw materials of W. cocos.

The Phosphatome of Medicinal and Edible Fungus Wolfiporia cocos.

Wolfiporia cocos is an important medicinal and edible fungus that grows in association with pine trees, and its dried sclerotium has been used as a traditional medicine in China for centuries. However, the commercial production of W. cocos sclerotia is currently limited by shortages in pine wood resources. Since protein phosphatases (PPs) play significant roles in growth, signal transduction, development, metabolism, sexual reproduction, cell cycle, and environmental stress responses in fungi, the phosphatome of W. cocos was analyzed in this study by identifying PP genes, studying transcript profiles and assigning PPs to orthologous groups. Fifty-four putative PP genes were putatively identified in W. cocos genome based on homologous sequences searching using BLASTx program against the Saccharomyces cerevisiae, Fusarium graminearum, and Sclerotinia sclerotiorum databases. Based on known and presumed functions of orthologues of these PP genes found in other fungi, the putative roles of these W. cocos PPs in colonization, hyphal growth, sclerotial formation, secondary metabolism, and stress tolerance to environment were discussed in this study. And the level of transcripts from PP genes in the mycelium and sclerotium stages was also analyzed by qRT-PCR. Our study firstly identified and functional discussed the phosphatome in the medicinal and edible fungus W. cocos. The data from our study contribute to a better understanding of PPs potential roles in various cellar processes of W. cocos, and systematically provide comprehensive and novel insights into W. cocos economically important traits that could be extended to other fungi.

[Habitat areas extracting of Poria cocos in Anhui based on "3S" technologies].

"Adversity" is one primary element that impacts the pharmacology components of authentic Chinese herbal medicine. Knowledge about "adversity" is a precondition of yield estimation, quality monitoring, location selection and the geo-herbalism protection. Used 3S(GIS, RS and GPS) technology to combine multi-source key ecological factors of "Anling", and used parasitic relationships between organisms to extracting its suitable region for the first time. Results showed that the "Anling" were mainly distributed in Dabie coteau. Suitable area amount to 36.8 km², Yuexi, Shucheng, Jinzhai and Qianshan which account for about 93.55% of whole congenial region. The first three accounts for about 80.82%. It was Yuexi that account for 1/3 above especially. Field investigation verify accuracy of extraction about 91.67%, which has confirmed it is feasible that using the relationship between parasitic host and parasitic to extract native environment of parasitic traditional medicine based on 3S technology.

Accumulation of biomass and four triterpenoids in two-stage cultured Poria cocos mycelia and diuretic activity in rats.

Poria cocos (Schw.) Wolf, an important medicinal and food fungus, is well known in East Asia. Due to growing market demand, long cultivation period, and consumption of pine trunk during cultivation, developing alternative methods for producing P. cocos and/or its active components is of interest. In the present study, the effects of different culture methods on biomass and accumulation of four triterpenoids were investigated. The ethanol extract of fermented mycelium (EFM) was orally administered to rats. Urine output and concentrations of electrolytes (Na+, K+, and Cl-) were measured. Our results showed that mycelia grew better under continuous shaking culture condition (7.5 g DW·L-1), and higher triterpenoid levels were accumulated in two-stage culture (112 mg·L-1, 2.03%). The optimal starting time of static culture for triterpenoid yield was 4th d after shaking culture. Single administration of middle and high dose of EFM significantly increased urine output, Na+ and Cl- excretion, and Na+/K+ ratio. These results suggested that ethanol extract of cultured mycelia showed significant diuretic activity in rats and two-stage culture of P. cocos could be an alternative way to produce mycelia and triterpenoids.

A Comprehensive and Comparative Study of Wolfiporia extensa Cultivation Regions by Fourier Transform Infrared Spectroscopy and Ultra-Fast Liquid Chromatography.

Nowadays, Wolfiporia extensa as a popular raw material in food and medicine industry has received increasing interests. Due to supply shortage, this species of edible and medicinal mushroom has been cultivated in some provinces of China. In the present study, cultivated W. extensa collected from six regions in Yunnan Province of China were analyzed by an integrated method based on Fourier transform infrared (FT-IR) spectroscopy and ultra-fast liquid chromatography (UFLC) coupled with multivariate analysis including partial least squares discriminant analysis (PLS-DA) and hierarchical cluster analysis (HCA) in order to investigate the differences and similarities in different origins and parts. In the tested mushroom samples, characteristic FT-IR spectra were obtained for acquiring comprehensive fuzz chemical information and pachymic acid was determinated as a biomarker in the meantime. From the results, the comparison of samples was achieved successfully according to their geographical regions and different parts. All the samples displayed regional dependence and the inner parts showed better quality consistency. In addition, the chemical constituents of cultivated W. extensa could be also affected by the cultivation methods. Meanwhile, there was an interesting finding that the soil properties of cultivation regions may have a relationship with the chemical constituents of the epidermis of soil-cultured W. extensa, rather than the inner parts. Collectively, it demonstrated that the present study could provide comprehensive chemical evidence for the critical complement of quality evaluation on the cultivated W. extensa. Moreover, it may be available for the further researches of complicated mushrooms in practice.

De novo assembly and transcriptome analysis of sclerotial development in Wolfiporia cocos.

Wolfiporia cocos Ryvarden et Gilbertson, a well-known medicinal fungus in the Basidiomycetes, is widely distributed in East Asia. Its dried sclerotium, which is known as Fuling in China, has been used as a traditional crude drug in Chinese traditional medicine for thousand years. However, little is known about how the sclerotium is developed at the genetic level. In this study, the de novo sequencing of sclerotia of W. cocos (S1_initial stage; S2_developmental stage and S3_mature stage) was carried out by illumina HiSeq 2000 technology. 27,438 unigenes were assembled from ~30Gbp raw data, and 12,093 unigenes were significantly annotated. The analysis of expression profiles during development returned 304 differentially expressed genes (DEGs), which were clustered into four different groups according to their expression trends. Especially for the maturation stage (S3), the sclerotium exhibited a markedly different expression profile from other stages. We further showed that peroxisome, unsaturation of fatty acids and degradation pathway were respectively prevalent in S1, S2 and S3 stages as evidenced by enrichment analysis. To our knowledge, this study represents the first report of sclerotial development transcriptomics in W. cocos. The obtained results provide novel insights into the developmental biology of the sclerotia, which is helpful for future studies about cultivation and breeding of W. cocos.

Transcriptome and Secretome Analyses of the Wood Decay Fungus Wolfiporia cocos Support Alternative Mechanisms of Lignocellulose Conversion.

UNLABELLED: Certain wood decay basidiomycetes, collectively referred to as brown rot fungi, rapidly depolymerize cellulose while leaving behind the bulk of cell wall lignin as a modified residue. The mechanism(s) employed is unclear, but considerable evidence implicates the involvement of diffusible oxidants generated via Fenton-like chemistry. Toward a better understanding of this process, we have examined the transcriptome and secretome of Wolfiporia cocos when cultivated on media containing glucose, purified crystalline cellulose, aspen (Populus grandidentata), or lodgepole pine (Pinus contorta) as the sole carbon source. Compared to the results obtained with glucose, 30, 183, and 207 genes exhibited 4-fold increases in transcript levels in cellulose, aspen, and lodgepole pine, respectively. Mass spectrometry identified peptides corresponding to 64 glycoside hydrolase (GH) proteins, and of these, 17 corresponded to transcripts upregulated on one or both woody substrates. Most of these genes were broadly categorized as hemicellulases or chitinases. Consistent with an important role for hydroxyl radical in cellulose depolymerization, high transcript levels and upregulation were observed for genes involved in iron homeostasis, iron reduction, and extracellular peroxide generation. These patterns of regulation differ markedly from those of the closely related brown rot fungus Postia placenta and expand the number of enzymes potentially involved in the oxidative depolymerization of cellulose. IMPORTANCE: The decomposition of wood is an essential component of nutrient cycling in forest ecosystems. Few microbes have the capacity to efficiently degrade woody substrates, and the mechanism(s) is poorly understood. Toward a better understanding of these processes, we show that when grown on wood as a sole carbon source the brown rot fungus W. cocos expresses a unique repertoire of genes involved in oxidative and hydrolytic conversions of cell walls.