Lactarius Hatsudake Research Articles

Anti-hepatocarcinoma effect of polysaccharides extracted from Lactarius hatsudake Tanaka mushroom: in vitro and in vivo studies

Anti-hepatocarcinoma effect of polysaccharides extracted from <em>Lactarius hatsudake</em> Tanaka mushroom: in vitro and in vivo studies

The Soil Bacterial Community Structure in a Lactarius hatsudake Tanaka Plantation during Harvest.

Lactarius hatsudake Tanaka is a mycorrhizal edible mushroom with an appealing taste and rich nutrition. It is also a significant food and has medicinal value. In this study, the plantation of L. hatsudake during the harvest period was taken as the research object, and this article explores which bacteria in the soil contribute to the production and growth of L. hatsudake. The soil of the control (CK) and the soil of the mushroom-producing area [including the soil of the base of the mushroom (JT) and the mycorrhizal root soil (JG)] was collected in the plantation. The three sites' bacterial community structure and soil diversity were analyzed using high-throughput sequencing technology, and a molecular ecological network was built. Soil bacteria in the L. hatsudake plantation had 28 tribes, 74 classes, 161 orders, 264 families, 498 genera, and 546 species. The dominant phyla were Proteobacteria and Acidobacteria, and the dominant genera were Burkholderia_Caballeronia_Paraburkholderia, Acidothermus, Bradyrhizobium, Candidatus_Xiphinematobacter, and Granulicella. The α-diversity of soil bacteria in JT was significantly lower than that in JG and CK, and the β-diversity in JT samples was significantly different from that in JG and CK samples. The size and complexity of the constructed network were smaller in JT samples than in JG and CK samples, and the stability was higher in JT samples than in JG and CK samples. The positive correlation between species in JT samples was dominant. The potential mycorrhizal helper bacteria (MHB) species of L. hatsudake was determined using correlation and differential group analysis. The results support future research on mycorrhizal synthesis, plantation management, and the function of microorganisms in the soil rhizosphere of L. hatsudake.

Glucan polysaccharides isolated from Lactarius hatsudake Tanaka mushroom: Structural characterization and in vitro bioactivities

Commercially available mushroom polysaccharides have found widespread use as adjuvant tumor treatments. However, the bioactivity of polysaccharides in Lactarius hatsudake Tanaka (L. hatsudake), a mushroom with both edible and medicinal uses, remains relatively unexplored. To address this gap, five L. hatsudake polysaccharides with varying molecular weights were isolated, named LHP-1 (898 kDa), LHP-2 (677 kDa), LHP-3 (385 kDa), LHP-4 (20 kDa), and LHP-5 (4.9 kDa). Gas chromatography-mass spectrometry, nuclear magnetic resonance, and atomic force microscopy, etc., were employed to determine their structural characteristics. The results confirmed that spherical aggregates with amorphous flexible fiber chains dominated the conformation of the LHP. LHP-1 and LHP-2 were identified as glucans with α-(1,4)-Glcp as the main chain; LHP-3 and LHP-4 were classified as galactans with varying molecular weights but with α-(1,6)-Galp as the main chain; LHP-5 was a glucan with β-(1,3)-Glcp as the main chain and β-(1,6)-Glcp connecting to the side chains. Significant differences were observed in inhibiting tumor cell cytotoxicity and the antioxidant activity of the LHPs, with LHP-5 and LHP-4 identified as the principal bioactive components. These findings provide a theoretical foundation for the valuable use of L. hatsudake and emphasize the potential application of LHPs in therapeutic tumor treatments.

Establishment of Pinus massoniana–Lactarius hatsudake Symbiosis

Lactarius hatsudake is a common ectomycorrhizal edible mushroom in Pinus massoniana forests, and has important ecological and potential economic values. However, there are only a few reports on the establishment of Pinus massoniana–Lactarius hatsudake symbiosis. Here, we isolated a new strain of L. Lactarius hatsudake (GX01) from a local masson pine forest and established its ectomycorrhizal symbiosis with the P. massoniana. Potato dextrose agar (PDA) medium was optimal for the growth of L. hatsudake GX01. The saffron-to-brown ectomycorrhiza formed by L. hatsudake GX01 are usually bifurcated or coralloid shape, with a rod and a smooth surface, without emanating hyphae. The characteristic mantle and Hartig net structures of ectomycorrhizae were confirmed by microscope and scanning electron microscope (SEM). L. hatsudake GX01 can significantly promote the formation and development of lateral roots of P. massoniana seedlings during the early interaction. This study thus lays the foundation for subsequent study of the symbiotic molecular mechanism and application of P. massoniana–L. hatsudake symbiosis.

Modulatory Effects of Lactarius hatsudake on Obesity and Gut Microbiota in High-Fat Diet-Fed C57BL/6 Mice.

Lactarius hatsudake (LH), a great wild endemic fungus, contains rich nutritional components with medicinal properties. The effects of LH on body weight, liver weight, liver injury, blood lipids, and gut microbiota in C57BL/6 mice fed a high-fat diet (HFD) for 8 weeks was examined in this research. Though there was no clear impact on weight loss, the findings indicate that LH treatment effectively decreased liver damage caused by HFD, as well as lowered serum total cholesterol, triacylglycerol, and low-density lipoprotein cholesterol levels. Additionally, it positively influenced gut microbiota to resemble that of mice on a normal diet. In HFD-fed mice, LH markedly boosted the levels of Parabacteroides, unclassified Muribaculaceae, Oscillibacter, and unclassified Oscillospiraceae, while reducing the abundance of Lachnospiraceae NK4A136 group and Erysipelatoclostridium, as well as the ratio of Firmicutes to Bacteroidetes. Further analysis of correlation indicate a possible connection between obesity and gut microbiota. Obesity-related indices show a positive association with unclassified Eubacterium coprostanoligenes group, Blautia, and Erysipelatoclostridium, while displaying a negative correlation with unclassified Muribaculaceae, unclassified Clostridia vadinBB60 group, Helicobacter, Oscillibacter, unclassified Ruminococcaceae, Parabacteroides, and unclassified Oscillospiraceae. The results suggest that LH can help combat obesity and may have the potential to be utilized as a functional food.

Diversity and Network Relationship Construction of Soil Fungal Communities in Lactarius hatsudake Tanaka Orchard during Harvest.

Lactarius hatsudake Tanaka is a mycorrhizal edible mushroom with rich economic and nutritional value. Although it is artificially planted, its yield is unstable. Soil fungi, including L. hatsudake, coexist with many other microorganisms and plants. Therefore, complex microbial communities have an influence on the fruiting body formation of L. hatsudake. L. hatsudake and its interactions with the rest of the fungal community over time are not completely understood. In this study, we performed high-throughput sequencing of microorganisms in the basal soil of the fruiting body (JT), mycorrhizosphere soil (JG), and non-mushroom-producing soil (CK) in a 6-year-old L. hatsudake plantation at harvest. The results showed that the soil of the L. hatsudake plantation was rich in fungal communities and a total of 10 phyla, 19 classes, 53 orders, 90 families, 139 genera, and 149 species of fungi were detected. At the phylum level, the major groups were Basidiomycota and Ascomycota. At the genus level, the dominant groups were Lactarius, Trichoderma, Suillus, and Penicillium. Among them, L. hatsudake had an absolute dominant position in the soil fungal community of the plantation, and was the only group of Lactarius in the plantation soil. Penicillium cryptum and Penicillium adametzii were unique to the JT soil sample. Chaetopsphaeria, Myxocephala, Devriesia, and Psathyrella were positively correlated with L. hatsudake. In the constructed fungal network, the total number of nodes were ranked in descending order as JG (441) > CK (405) > JT (399), while the total number of edges were ranked in descending order as CK (1360) > JG (647) > JT (586). Analysis of the fungal assembly process revealed that groups CK and JG have determinative processes that dominated community building, while the JT group exhibited a dominant random process with a 0.60 probability. The results indicated that L. hatsudake was successfully colonized in the plantation soil. During harvest, the CK group exhibited the largest network size and the most complex fungal interactions, while the fungal community structure in the mushroom cultivation zone (JT and JG) was stable and less susceptible to external environmental interference. L. hatsudake affects the fungal community in the soil surrounding its fruiting body.

Nutritional value, elemental bioaccumulation and antioxidant activity of fruiting bodies and mycelial cultures of an unrecorded wild Lactarius hatsudake from Nanyue mountainous region in China

An unrecorded wild mushroom Lactarius hatsudake from Nanyue mountainous region in China was identified. Subsequently, comparative investigation on the nutritional value, elemental bioaccumulation, and antioxidant activity was performed in the fruiting body (FB) and mycelium (MY) samples of this species. It revealed that the contents of moisture (87.66 ± 0.16 g/100 g fw) and ash (6.97 ± 0.16 g/100 g dw) were significantly higher in FB, and the total carbohydrate, fat, and protein concentrations of FB were similar to those in MY. Among nutritionally important elements, FB possessed higher concentrations of potassium (37808.61 ± 1237.38 mg/kg dw), iron (470.69 ± 85.54 mg/kg dw), and zinc (136.13 ± 5.16 mg/kg dw), whereas MY was a better source of magnesium (1481.76 ± 18.03 mg/kg dw), calcium (2203.87 ± 69.61 mg/kg dw), and sodium (277.44 ± 22.93 mg/kg dw). According to the health risk estimation, FB might pose an aluminum-related health problem when a prolonged period of exposure, while MY was risk-free for consumers. The results of antioxidant capacity (1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assays) in FB and MY were within the range of 104.19 ± 5.70 mg ascorbic acid equivalents (AAE)/g to 169.50 ± 4.94 mg AAE/g, and half maximal effective concentration EC50 values ranged from 0.23 ± 0.01 mg/mL to 0.62 ± 0.05 mg/mL. The aqueous extracts of MY demonstrated a strong ABTS radical scavenging capacity with the highest AAE value.

A high-quality genome assembly of Lactarius hatsudake strain JH5.

Lactarius hatsudake is a species of Lactarius commonly found in pine forests, is edible with a delicious and nutritious fruiting body, and exhibits medicinal properties. It is an ideal natural multifunctional food with bioactive components including fungal polysaccharides, crude fiber, unsaturated fatty acids, nucleic acid derivatives, various amino acids, and vitamins. However, biological and genomic analyses of this mycorrhizal mushroom are sparse, thereby hindering large-scale cultivation. Previously, we isolated and screened L. hatsudake JH5 strains and have applied our garnered knowledge to the large-scale cultivation of mycorrhizal seedlings. In this study, we produced a high-quality genome assembly of L. hatsudake JH5 by combining Illumina paired-end and PacBio single molecule real-time sequencing, resulting in PacBio single molecule real-time reads of 7.67 Gb and Illumina Pair-End reads of 1,560 Mb. Based on the distribution of k-mer frequencies, the genome size of this strain was estimated to be 63.84 Mb (1.14% heterozygosity). Based on de novo genome assembly, the final genome size was determined to be 76.7 Mb, with scaffold N50 of 223.2 kb and N90 of 54.5 kb, and a GC content of 54.38%. BUSCO assessment showed that genome completeness was 89.0%. The N50 length of the JH5 genome was 43.6% longer than that of the previously published L. hatsudake MG20 genome. This high-quality L. hatsudake genome assembly will facilitate research on the functional genome, molecular breeding, yield enhancement, and sustainability of L. hatsudake cultivation.

Phenolic Acid Profiling of Lactarius hatsudake Extracts, Anti-Cancer Function and Its Molecular Mechanisms.

Cancer is still the leading cause of death across the world, and there is a lack of efficient therapies. Lactarius hatsudake is a mushroom with a food and medicine homology that contains numerous biologically active substances. This study aimed to investigate the composition of extracts from Lactarius hatsudake (L. hatsudake) and their anti-cancer function and molecular mechanisms. Our results showed that the total phenolic content of L. hatsudake extracts was 139.46 ± 5.42 mg/g. The following six phenolic compounds were identified from L. hatsudake extracts by HPLC and UPLC-QTOF/MS: gallic acid, pyrogallol, chlorogenic acid, ferulic acid, myricetin, and cinnamic acid. Colorectal cancer cell HCT116 and hepatic cancer cell HepG2 were used to evaluate the anti-cancer function of the L. hatsudake extracts. Compared with HepG2 cells, the L. hatsudake extracts showed stronger anti-cancer activity against HCT116 cells and these were used to study molecular mechanisms. The results indicated that the L. hatsudake extracts could arrest the cancer cell cycle and inhibit cancer cell proliferation, which may be mediated by the MAPK/NFκB/AP-1 signalling pathway; the L. hatsudake extracts also promoted cancer cell apoptosis through a mitochondrial-dependent pathway. Taken together, these findings demonstrate that L. hatsudake ethanol extracts contain six main phenolics and illustrate the remarkable potentiality of L. hatsudake as a source of natural phenolics for cancer prevention and as an adjuvant in the treatment of functional foods.

Novel Fluorescent Nano Carbon Quantum Dots Derived From Lactarius Hatsudake for High Selective Vitamin B12 Detection.

VB12 is involved in the production of bone marrow red blood cells, preventing pernicious anemia and brain nerve damage. A large edible fungus Lactarius hatsudake (Lh) was used as a carbon source to produce fluorescent carbon dots (CDs) with high quantum yield and high photostability by a one-step hydrothermal method. The structure and optical properties of Lh-CDs were characterized by XRD, XPS, FTIR, and UV-Vis, and the stability of Lh-CDs was studied. The maximum excitation wavelength and emission wavelength of Lh-CDs were 324 and 408 nm, respectively, and the average fluorescence lifetime was 2.29 ns. The linear range is 0∼20 μM (R2 = 0.9972) with a detection limit of 36.9 nM. At the same time, the mechanism of fluorescence quenching was internal filtration effect. It showed that Lh-CDS had good selectivity, high sensitivity, and good anti-interference ability to VB12, which provided a new detection method for VB12.Highlights The fluorescence intensity of the LH-CDs could be effectively quenched by VB12. The fluorescence intensity of the LH-CDs could be effectively quenched by VB12.

An acidic feruloyl esterase from the mushroom Lactarius hatsudake: A potential animal feed supplement

An acidic feruloyl esterase (lhFAE) from Lactarius hatsudake was purified to homogeneity, resulting in a high specific activity of 10.3U/mg toward methyl ferulate as substrate, which was 33.2-fold higher than that of the crude enzyme. lhFAE was a monomeric protein with a molecular mass of 55kDa. All three internal amino acid sequences exhibited 100% identity with the Laccaria bicolor carbohydrate esterase family 9 protein. The enzyme demonstrated high activity toward methyl ferulate (MFA) at an optimum pH of 4.0 and an optimum temperature at 30°C, and under these conditions, the Km and Vmax were 0.54mM and 13.84Umg−1, respectively. The purified FAE exhibited preference for methyl caffeate over methyl ferulate, and was least active on methyl p-coumarate. Interestingly, above 80% of the enzyme activity was preserved in the presence of metal ions at 5.0mM concentration, which indicated that lhFAE was highly metal-tolerant. lhFAE was able to release ferulic acid from de-starched wheat bran (DSWB) both in the absence and presence of xylanase, and 70.2% (of the total amount of) ferulic acid (FA) was released by the synergistic action of xylanase. The result showed that lhFAE has good acid and metal ion stability, and is suitable to be used as an animal feed supplement.

English

Bio-fertilizers are of great importance in forestry because they can positively affect plant growth and the environment. In this study, the seedlings of slash pine (Pinus elliottii) were planted in pots. One month later they were inoculated with 7 groups of ectomycorrhizal fungi including&nbsp;Lactarius&nbsp;sp.,&nbsp;Scleroderma polyrhizum&nbsp;Pers.,&nbsp;Lactarius&nbsp;hatsudake&nbsp;Tanaka,Lycoperdon&nbsp;sp.,&nbsp;Lycoperdon&nbsp;sp./&nbsp;Lactarius&nbsp;sp.,&nbsp;Lycoperdon&nbsp;sp. /&nbsp;Lactarius hatsudakeTanaka and&nbsp;Scleroderma polyrhizum&nbsp;Pers./&nbsp;Lactarius&nbsp;sp.. Plant growth variables were determined after three months. The results showed that: (1) all inocula significantly increased the ground diameter and plant height and strengthened the overall absorption of nitrogen, phosphorus and potassium; (2) the best inocula mixture was&nbsp;Lycoperdon&nbsp;sp. /Lactarius hatsudake&nbsp;Tanaka, likely due to synergistic interactions between the two partners and (3) mixed ectomycorrhizal fungi were not necessarily better than single fungus. &nbsp; Key words:&nbsp;Slash pine (Pinus elliottii) growth, ectomycorrhizal inocula, bio-fertilizers, interaction, mixed ectomycorrhizal fungi.

Analysis of genetic diversity of Lactarius hatsudake in South China

Lactarius hatsudake is a type of ectomycorrhizal fungus that significantly influences the growth of pine trees. It is widely prevalent in Asian countries and has a high economic value. Artificial cultivation of this fungus has not been achieved as yet; therefore, excessive manual harvesting may cause serious damages to the site of its production. In this study, we analyzed 41 samples of L. hatsudake from south China using internal transcribed spacer (ITS) sequences. By comparing the differences among ITS sequences to identify the haplotype diversity within each population, the relationships among local populations, the relationship between the level of genetic differentiation and geographical separation, and the contributions of local and regional geographical separations to the overall ITS haplotype variation were analyzed. Genetic analysis indicates that ITS sequences obtained from these 41 L.hatsudake samples could be identified as 18 haplotypes, of which 13 haplotypes were contained in only a single sample, whereas the remaining sequence types all were contained in two or more samples. The most common sequence type, haplotype 6, was found in 16 samples and was distributed across nearly every region. The Mantel test demonstrated that there is no significant linear relationship between geographical distance and the F(ST) value of genetic difference. Results of this research illustrates that there exists a certain degree of genetic intermixing among natural populations of L. hatsudake. From the group genetic analysis, it appears that there exists genetic differentiation of lower frequencies in natural populations of L. hatsudake; however, the linear relationship between the degree of genetic differentiation and geographical distance is not distinctly apparent.

Investigation and Screening of Superior ECM Fungi Strains for Plantation in South China

The ectomycorrhizal fungi resource ofPinus massonianaandChinese Firforest had been investigated in the places of Hunan and YunNan Province. There were 17 species belong to 6 families and 9 genera. All the collected ectomycorrhizal fungi were separated by tissue isolation method, and 12 strains were obtained, which belong to 7 genera of 4 families. The preliminary selection results showed that there were 6 strains presented the better culture characteristics, which wereSeleroderma polyrhizum Pers.,Scleroderma cepa Pers.,Calvatia craniiformis(Schw.)Fr.,Lactarius hatsudake Tanaka,Lactarius deliciosus(Fr)Gray.,Russula puellaris Fr.. Then these 6 strains were cultivated between every two among them, the results showed that the group of LS3/RHG1 was the optimal combination, the next were the groups of LGD-2/La-HZ3 and LS3/La-HZ3. The groups of A3/ La8 showed antagonism to each other, so it was not suitable to be combined for inoculation. These research results will provide the important technical support for expanding propagation of ECM fungi and mycorrhizal technology application inPinus massonianaandChinese Firplantation.

ChemInform Abstract: Lactariolines A and B: New Guaiane Sesquiterpenes with a Modulatory Effect on Interferon‐γ Production from the Fruiting Bodies of Lactarius hatsudake.

AbstractThe two new guaiane sesquiterpenes lactarioline A (I) and B (II) inhibit IFN‐γ production in NK92 cells in a dose‐dependent manner.

Character impact odorants from wild mushroom (Lactarius hatsudake) used in Japanese traditional food

AbstractThe components of the volatile oil from wild mushroom (Lactarius hatsudake), used in Japanese traditional food, were analysed and quantified for the first time by capillary GC and GC–MS. Seventy‐six components were separated from the oil and of these 71 components were identified. The main components of the oil were oxidized sesquiterpenes [cis‐isolongifolanone (624.9 μg/100 g), α‐cedrene epoxide (578.7 μg/100 g), humulene epoxide III (453.9 μg/100 g), clovane (425.4 μg/100 g)], aliphatic acids [linoleic acid (585.9 μg/100 g) and palmitoleic acid (333.3 μg/100 g)]. Odour evaluation of the volatile oil from L. hatsudake was also carried out using GC–MS/olfactometry (GC–MS/O) and aroma extract dilution analysis (AEDA), from which it was found that hexanal, 4‐dehydroviridiflorol, myliol and phenylacetaldehyde seem to contribute to the green, spicy and sweet odour of L. hatsudake. Copyright © 2010 John Wiley &amp; Sons, Ltd.

Lactariolines A and B: new guaiane sesquiterpenes with a modulatory effect on interferon-γ production from the fruiting bodies of Lactarius hatsudake

Lactariolines A and B: new guaiane sesquiterpenes with a modulatory effect on interferon-γ production from the fruiting bodies of Lactarius hatsudake

Bioactive ergosterol derivatives isolated from the fungus Lactarius hatsudake

Bioactive ergosterol derivatives isolated from the fungus Lactarius hatsudake

Ergosterol peroxides as phospholipase A2 inhibitors from the fungus Lactarius hatsudake

Four ergosterol derivatives (1-4) have been isolated for the first time from the fruiting bodies of a basidiomycete fungus, Lactarius hatsudake, through activity-guided fractionation. Their structures were determined, using spectroscopic analysis, as: (22E,24R)-ergosta-5,7,22-dien-3beta-ol (ergosterol, 1); 5alpha,8alpha-epidioxy-(22E,24R)-ergosta-6,22-dien-3beta-ol (ergosterol peroxide, 2); 5alpha,8alpha-epidioxy-(24S)-ergosta-6-en-3beta-ol (3); and (22E,24R)-ergosta-7,22-dien-3beta,5alpha,6beta-triol (cerevisterol, 4). Compounds 2 and 3 showed selective inhibitory activity against Crotalus adamenteus venom phospholipase A(2) (PLA(2)) enzyme, but not against Apis mellifcra bee venom PLA(2). The antiphospholipase A(2) activity of compounds 2 and 3 are reported here for the first time.

Two New Azulene Pigments from the Fruiting Bodies of the BasidiomyceteLactarius hatsudake

Two new azulene pigments, 7-(1-hydroxy-1-methylethyl)-4-methylazulene-1-carbaldehyde (1) and 4-methyl-7-(1-methylethyl)azulene-1-carboxylic acid (2), were isolated from the fruiting bodies of the basidiomycete Lactarius hatsudake, together with one known azulene pigment (3). Their structures were determined by spectroscopic means (including 2D-NMR) and by HR-TOF-MS experiments.