The first record on the medicinal use of “Sanghuang” date back 2 000 years to China’s The Divine Farmer’s Materia Medica, by the name “Sanger”. The term “Sanghuang” was first reported in a Chinese medicinal book The characters of drugs written by Zhen Quan in early Tang Dynasty. Sanghuang differs from other medicinal fungi in having many morphologically resembling species. The “Sanghuang” appeared in the numerous ancient books has been described by different people and actually represents different species. In ancient times it was not possible to study both microscopic morphological features and DNA sequencing, to clarify species separation. Modern research of Sanghuang originated from discovery of its excellent anti-tumor capability by the Japanese scholars in 1968. In Japan and Korea, Phellinus linteus was long adopted as the scientific binomial for the Sanghuang species. However, in 1998 the Chinese mycologists found P. linteus is distributed in Central America, not in Asia. In 2012, the genuine Sanghuang was presented as a new species, Inonotus sanghuang, grows only on Morus. In 2016, the new genus Sanghuangporus was proposed to accommodate I. sanghuang and some other closely related species, and Sanghuangporus sanghuang thus became the scientific binomial for the Sanghuang species. So far, Sanghuangporus comprises 14 species, which are generally specialized with their host tree species, and S. sanghuang is the only species grows on Morus. Pharmacological activities of S. sanghuang are better than two other commercially available species, S. baumii and S. vaninii. The cultivated fruiting bodies of the so-called Sanghuang in China, Japan and Korea, are not S. sanghuang, but S. vaninii. According to the excellent health-care effects and safety of S. sanghuang and S. vaninii, we suggest the government departments accept these two medicinal fungi in the China Pharmacopoeia, and consider the fungi as safe food stuff and Chinese herbal medicine for promoting development of the Sanghuang industry. It is essential to clearly specify the correct scientific name and Chinese name usages of these two medicinal fungi used in their products.
Vernicia fordii (tung oil tree), an important industrial oil crop, usually suffers from a devastative disease caused by soil-borne Fusarium oxysporum f. sp. fordiis (Fof-1), and the effectiveness of chemical control for this Fusarium wilt disease is undesirable till now. It was found that the richness of Trichoderma species in rhizosphere soil of wilt-resistant tung oil trees was relatively high. In this study, 16 strains of Trichoderma were isolated from the rhizosphere soil samples of the wilt-resistant trees, and four strains antagonistic to F. oxysporum, T. koningiopsis TkonT1, T. spirale TspiT2, T. atroviride TatrT3 and T. harzianum TharT4, were screened. Microscopical observation proved that TkonT1, TharT4 and TspirT2 were confirmed to have better antifungal effect. TkonT1 and TatrT3 could wrap or penetrate the hyphae of Fof-1 and absorb nutrients from host, resulting in hyphal disruption and destruction. The results indicated that the Trichoderma species from wilt-resistant tree rhizosphere soil could be used for biocontrol of Fusarium wilt disease of V. fordii trees.
Dark septate endophytes (DSE) are extensively inhabiting roots of a wide range of plants and play vital roles in promoting plant growth and improving plant performance under various adverse environments. In this study, a DSE strain was isolated from Pinus massoniana root and its symbiotic characteristics with P. massoniana were investigated under aseptic conditions. Morphological and molecular biological evidence indicated that the DSE isolated was Phialocephala fortinii. Under pure culture condition, P. fortinii does not produce spores, and its septate mycelium is melanized with septa. PDA medium is most suitable for its growth. Three periods emerged during the symbiosis establishment between P. fortinii and P. massoniana. In the pre-infection phase, the newly emerged hyphae grew towards and started attaching the root in 2 days of post inoculation (dpi), but without hyphal penetration. During the infection phase, the mycelium began penetrating into the root of P. massoniana, and the intraradical mycelium extended radially along the intercellular space of cortical cells in 4 dpi; in 6 days of post-inoculation, the mycelium continued to expand in the roots and formed intensive microsclerotia filled with lipids. P. fortinii inoculation significantly increased the biomass of P. massoniana (P<0.05) and modified root development. The growth of the primary root was hampered, while the growth of the lateral root significantly increased (P<0.05). The length of the lateral root increased by 112.87% as compared with the non-inoculation treatment. Root hairs in the infected roots were bound by intensive extraradical mycelium. The results provide reference for further investigation of the symbiotic mechanisms between DSE and its host.
Overexpression of multidrug resistance (MDR) gene in tumor cells is an internal cause of drug resistance of tumor cell and is the leading cause of failure in tumor chemotherapy. Therefore, making a search for an inhibitor of MDR activity is an effective way to increase the anti-tumor drug potency. In this study, drug-resistant cell line of human breast cancer cell line (MCF-7) was established by a low-level continuous induction method. The results showed that MCF-7 drug-resistant cell line has cross-resistance to doxorubicin (ADM), paclitaxel and cisplatin with the indexes (resistance index, RI) of 5.11, 3.55 and 1.79 respectively. Screening of tumor cell MDR reversal activity showed that Tubeufia rubra PF02-2 and T. hechiensis XSL05 had MDR reversal activity with reversion folds (RF) of 3.79 and 1.07 respectively. These results indicate that T. rubra and T. hechiensis have potential to be developed as MDR inhibitors.
Morchella importuna is a kind of rare edible fungi, which can be cultivated in field covered with soil. The contamination of heavy metals in soil is becoming increasingly serious, constituting a potential threat to the mycelial growth as well as the safety and quality of fruiting bodies of M. importuna. RT-PCR dection indicated that the expression of candidate gene ATX1 was significantly down-regulated on condition that the mycelium of M. importuna was treated with cadmium stress. The gene ATX1 was cloned and the domain of MiATX1p was predicted. It was found that MiATX1p might be related to copper transport and heavy metal stress. ATX1 over-expression and RNAi gene-silencing vectors were constructed separately. Agrobacterium- mediated transformation method was used to transfer ATX1 RNAi gene-silencing and over-expression vectors into the homokaryotic strain A50 of M. importuna. Four ATX1 silenced transformants with significantly down-regulated ATX1 expression and four RNAi gene-silencing transformants with significantly up-regulated ATX1 expression were screened respectively. Cadmium sensitivity tests indicated that ATX1 silenced transformants showed increased cadmium resistance, while ATX1 over-expression transformants showed decreased cadmium resistance. These results suggested that the expression of ATX1 gene was negatively correlated with cadmium resistance of M. importuna. ATX1p may play an important role in the response of M. importuna to cadmium stress.
Shake flask culture was used to study the effects of different concentration of Cu(II) on the morphological development and ligninase activity of Fusarium sp. ZH-H2. The results showed that when Cu(II) concentration was 0.50mmol/L, the highest mycelial biomass was up to 0.38g, and the mycelia were white, smooth and spherical and reached the maximum in number. At this concentration, the peak value of lignin peroxidase (LiP) activity reached 28 351.41U/L. Cu(II) concentration of 2.00mmol/L was beneficial to induce the metabolism of manganese peroxidase (MnP) and laccase (Lac). The highest peak value of MnP and Lac were 1 612.90U/L and 7 829.12U/L, respectively, and the peak formation time was significantly shortened. Excessively high or low concentration brought about inhibitory effect on the activity of the three lignin enzymes.
Artificial domestic cultivation of a wild strain of Agrocybe praecox collected from reedy wetland in the Datong Area of Dongting Lake, Hunan Province, is carried out. Reed shavings were mainly used as substrate. Using liquid spawn and soil-covering cultivation method, average bioconversion rates of the first-season mushrooms was 45.6%. Nutrient analysis showed that using reed shavings as substrate, the content of main mineral elements in fruiting bodies was K 2 190mg/kg, Fe 16.2mg/kg, Mg 59.7mg/kg and Ca 33.4mg/kg. The mushrooms were rich in glutamic acid, methionine and aspartic acid. The proportion of essential amino acid content was up to 44.85%. Agrocybe praecox is a kind of edible fungus with high protein, low fat, high potassium and low sodium.
Degradation of lignocellulose by Trichoderma viride, a high cellulase-producing fungus, is an effective method for recycling waste materials. Two strains of T. viride were isolated respectively from rhizosphere soil of Glycyrrhiza uralensis and Saposhnikovia divaricata in different habitats. The FPase activity and reducing sugar production of the strains growing on two lignocellulosic substrates, i.e. corn stalk and liquorice residue were assessed. The environmental adaptability and growth state of both strains on the two different substrates were observed in terms of different fermentation time, incubation temperatures, solid to liquid ratios and initial pH. The results showed that the optimum initial solid to liquid ratio of corn stalk substrate for T. viride XJ was 1:4-1:5.5, and that for T. viride AG was 1:5-1:5.5. The FPase activity of T. viride AG was significantly higher than that of T. viride XJ in the substrate with initial solid to liquid ratio of 1:5.5. The optimal fermentation temperature of both strains was 28°C. The FPase activities significantly varied with tested temperatures, but showed no difference between T. viride XJ and T. viride AG. The reducing sugar was consumed by the two T. viride strains. Using liquorice residue as substrate, the optimum initial solid to liquid ratio was 1:2-1:2.5 for T. viride XJ, and 1:3-1:3.5 for T. viride AG. The FPase activity of T. viride AG was significantly higher than that of T. viride XJ in the substrate with initial solid to liquid ratio over 1:3. The optimal fermentation temperature of T. viride AG was 28°C, and that of T. viride XJ was 23-28°C. The FPase activity of T. viride XJ was significantly higher than that of T. viride AG at temperatures below 28°C. The reducing sugar was accumulated by the two T. viride strains. The optimum initial pH for cultivation of both strains was 6-7, and the optimal fermentation time was 3 days. The FPase activity of T. viride AG was higher than that of T. viride XJ under optimal fermentation conditions. The liquorice residue was better than corn stalk for inducing FPase production by T. viride. The variation partitioning indicated that the differences of FPase activity between tested strains depended on the ecological adaptability of the strains.
The influence of arbuscular mycorrhizal fungi (AMF) Claroideoglomus etunicatum, Funneliformis mosseae and Gigaspora margarita on shade tolerance of Tagetes patula plants were determined under the conditions of different light intensity (shading rates of 0%, 24%, 48%, 72%, 96%). Results showed that the tested AMF could colonize roots of T. patula to form typical arbuscular mycorrhiza, and the best infection effect was achieved by inoculating with F. mosseae under different shading treatments, and the strong light and weak light were unfavourable to the infection of AMF. Under shading rate of 24%, the growth of T. patula was the best. Compared with the control treatment (without inoculation), the plant height, stem thickness, total leaf area, root/shoot ratio, specific leaf weight, flower number and flower diameter of T. patula were significantly increased after inoculation of F. mosseae. Furthermore, florescence was prolonged and root activity, chlorophyll-a, chlorophyll-b, total chlorophyll and soluble sugar content were heightened, the content of proline and the light compensation point decreased, and the light saturation point and the largest photosynthetic rate increased. In conclusion, proper shading was beneficial to the growth and development of T. patula, and AMF could enhance the adaptability of T. patula to light, promote the growth and development, reduce the damage caused by weak light, and enhance shade-endurance of the plants. Inoculation of F. mosseae was proved to be the most effective.
Sporoderm-broken Ganoderma lingzhi spore powder was extracted fractionally with soxhlet extraction method using petroleum ether, ethyl acetate, acetone, ethanol, methanol and water as solvents with different proper polarity, and six extracts were obtained. The extracts were analyzed with gas chromatography-mass spectrometry (GC/MS) and ultra-performance liquid chromatography coupled with electrospray time-of-flight/mass spectrometry (UPLC-QTOF/MS). A total of 101 compounds is identified, including 10 acids, 40 esters, 7 alcohols, 6 ketones, 2 phenols, 18 hydrocarbons, 9 steroids and 9 heteroatom- containing compounds. 40 compounds are detected using UPLC-QTOF/MS, including 1 sesquiterpene, 1 diterpenoid, 9 triterpenes, 4 alkaloids, 7 amides, 9 organic acids and 9 other compounds. Only 1 compound is in common among those obtained by GC/MS and UPLC-QTOF/MS analyses. 105 compounds strickly present in their own selective extract and not reappear in other unselective one, but there are 31 compounds which can simultaneously present in two or more extracts. The method is proved to be better for separating the multi-component composition and widening the detection range of the compounds in Ganoderma lingzhi spore powder.
Immune receptors of Sparassis latifolia β-D-glucan affecting macrophage RAW264.7 were determined, and immunomodulatory mechanism of the β-D-glucan was explored. MTT assay was used to measure the proliferation activity of macrophage RAW264.7 under different concentrations of S. latifolia β-D-glucan and the concentration with strongest ability to promote macrophage proliferation was screened. Macrophage RAW264.7 was treated with selected optimal concentrations of β-D-glucan; TLR4 antibody and TLR2 antibody were applied to act on macrophage RAW264.7 for 1 hour, and then the macrophages were cultured with β-D-glucan-containing cell culture solution. Cell culture supernatants were collected, and the production of NO, IL-6, TNF-α, IFN-β were detected. TLR4 mRNA expression in macrophages was measured by RT-PCR to extract total RNA from cells. Total macrophage protein was extracted and TLR4 protein expression was determined by western blot. The results indicated that S. latifolia β-D-glucan can significantly promote the proliferation of RAW264.7 macrophages, increase the production of NO, IL-6, TNF-α and IFN-β, and enhance TLR4 mRNA expression and protein expression (P<0.01). After treating with TLR4 antibody, the production of NO, IL-1β, TNF-α, and IFN-β were significantly lower in comparison with untreated groups (P<0.01). After treating with TLR2 antibody, the production of NO, IL-6, TNF-α, and IFN-β decreased, but the difference was not significant. S. latifolia β-D-glucan can activate the signal transduction pathway through the cell surface receptor TLR4, enhance the release of downstream cytokines, and thereby regulate the immune function of macrophage RAW264.7. TLR2 maybe not the immune receptor of S. latifolia β-D-glucan.
Floccularia luteovirens (=Armillaria luteovirens), is a precious edible medicinal fungus in China. In this study the analgesic, anti-inflammation and anti-migraine activities of the water extract of F. luteovirens (FLW) were evaluated using nitroglycerin (NTG) induced migraine of rats as animal model. Rats were orally administered various doses of FLW. After nitroglycerin induction, their typical symptoms were observed and the levels of NO, interleukin-6 (IL-6), interleukin-1β (IL-1β), 5-hydroxytryptamine (5-HT), norepinephrine (NE) and dopamine (DA) in plasma and brain were assessed. The results showed that the levels of NO, IL-6 and IL-1β were significantly reduced in serum after FLW treatment, and 5-HT, NE and DA levels in brain were advanced. FLW showed analgesic and anti-inflammatory effects in migraine rats.
Tobacco stems are the important by-products in the tobacco industry. In this study, the tobacco stems were pretreated with laccase produced by white-rot fungus Pycnoporus sanguineus to improve the quality of cigarette products incorporated with tobacco stems. Using Box-Behnken design, the degradation rates of lignin, cellulose, hemicellulose and pectin were adopted as the response values to build the equation models and optimize the conditions for pretreatment of tobacco stems by complex enzymes consisting of laccase, cellulase, hemicellulase and pectinase. The results showed that the best pretreatment to improve the commodity quality of cigarettes is addition of 30U laccase per 100g tobacco stems. Under the condition of material to liquid ratio 35%, 30°C and pH 5, the degradation rates of lignin, cellulose, hemicellulose and pectin in tobacco stems in 48h were 20.16%, 15.10%, 7.20% and 12.40%, respectively. For obtaining the similar degradation rate, the conditions optimized by response surface method were addition of 14.72U laccase, 1.00U cellulase, 1.00U hemicellulase and 8.45U pectinase per 100g tobacco stems. There was no significant difference between the measured values and the theoretical values of the degradation rate of each component of tobacco stems. Under the treatment of complex enzymes, the fine structure of the tobacco stem surface was destroyed and cavities were increased significantly. It was proved that the quality of cigarettes incorporated with tobacco stems pretreated with laccase produced by P. sanguineus was effectively improved, and the laccase complex enzymes could significantly reduce the applying quantity of laccase and thereby reduced the cost of production.
A method for detection of Russula subnigricans was established based on Taqman-MGB probe of real-time fluorescent quantitative PCR. A pair of primers and a Taqman-MGB probe were designed and synthesized according to the ITS sequences of R. subnigricans and its related species, and the common toxic Russula species were used for verification. The results showed that the primers had good specificity, and only R. subnigricans exhibited fluorescence signal. It just took 2h to complete the whole detection process. It was indicated that this method could provide a technical support for rapid detection of toxic mushroom poisoning.
Liquid fermentation of Cordyceps militaris was carried out using chinese medicine magnoliae officinalis cortex as additional substrate. Based on single factor experiment, the key influencing factors of the fermentation process were optimized by Box-Behnken experimental design. The results showed that the extracellular polysaccharide content and mycelial biomass of C. militaris liquid fermentation products were 3.11mg/mL and 18.81mg/mL, respectively, under the addition of magnoliae officinalis cortex of 5g and optimized fermentative condition including inoculum volume of 15.5mL, fermentation temperature of 25°C and fermentative duration of 9d. It was indicated that the content of extracellular polysaccharide in fermentation broth was obviously affected by inoculum quantity, and the biomass of mycelia was mainly affected by fermentation temperature. The optimized process is feasible, short-cyclic and controllable.
In the process of mycorrhiza synthesis of some edible species of Lactarius and Tuber in seedlings of Pinus and Quercus, nine species of exogenous competitive mycorrhizal fungi (ECMF) were found to contaminate the host plants. Identification result indicates that these ECMF belong to five genera, Amphinema, Rhizopogon, Scleroderma, Sphaerosporella and Suillus, in five families, Atheliaceae, Rhizopogonaceae, Sclerodermataceae, Pyronemataceae and Suillaceae. Species of Amphinema, Rhizopogon and Suillus mainly contaminated Pinus seedlings, and Quercus seedlings were mainly contaminated by Scleroderma. Sphaerosporella species could contaminate both Pinus and Quercus seedlings. Morphological characteristics of these ECMF were described.
