Effect of Alcohol Extract of Spore Powder of Lasiosphaera Calvatia on Wound Healing of Full-thickness Skin Defect in Mice / 中国实验方剂学杂志

ObjectiveTo compare the effect of different solvent extracts of spore powder and fruiting body of Lasiosphaera Calvatia on fibroblasts and wound healing of full-thickness skin defect， in order to screen the optimal medication part of Lasiosphaera Calvatia. MethodThe effect of water extract and alcohol extract of spore powder and fruiting body on cell proliferation and cell migration of mouse skin fibroblasts （MSF） were examined in vitro. Cell proliferation and activity test （CCK-8） method was used for cell proliferation， scratch assay was used for cell migration， flow cytometry was conducted to explore cell cycle， enzyme-linked immunosorbent assay （ELISA） was used to determine the production of collagen Ⅰ and Ⅲ. At the same time， a full-thickness skin defect wound model was established to investigate the therapeutic effect of different solvent extracts of spore powder. Ultraviolet-visible spectrophotometry was used to measure the contents of index components in different solvent extracts. ResultThe water extract of spore powder and fruiting body had certain cytotoxicity， while the alcohol extract could promote proliferation， migration and production of collagen Ⅰ and Ⅲ of MSF， and the effect of spore powder was significantly higher than that of fruiting body. When the concentration was 10 mg·L-1， the cell proliferation rate of alcohol extract of spore powder was as high as （159.22±15.95）%， and could promote MSF from the G0/G1 phase to S phase and G2/M phase with an increased proliferation index. The alcohol extract also promoted the migration of fibroblasts， secreted collagen Ⅰ and Ⅲ. On in vivo model， the alcohol extract of spore powder significantly accelerated wound healing on mice， effectively promoted the complete epithelialization of wound tissue， and generated new collagen fiber. The results of determination showed that the contents of polyphenols and flavonoids in the alcohol extract were higher than the alcohol extract of fruiting body. ConclusionThe alcohol extract of spore powder in Lasiosphaera Calvatia has active components in the treatment of wounds with good development prospect， and the medicinal components may be polyphenols and flavonoids.

Evaluation of hybrids and single spore isolates of paddy straw mushroom Volvariella volvacea (Bull.) Singer for fruiting body yield and nutritional quality

Aim: Developing high yielding single spore isolates and hybrid strains of paddy straw mushroom (Volvariella volvacea) with superior nutritional composition of the fruiting bodies.Methodology: Two contrasting high yielding strains (DMRO-463 and DMRO-484) of V. volvacea previously released for higher fruiting body yield were used for isolation of single spore isolates (SSIs). The slow growing SSIs were used for developing the hybrid strains by mating on Malt Extract Agar Medium Petri dishes. Conversely, the fast growing SSIs and the developed hybrid strains were screened for downward mycelial growth on paddy straw filled in wide mouth test tubes. Based upon downward mycelial growth, only eight SSIs and ten hybrid strains with numerically higher downward mycelial growth compared to two parents' checks were further evaluated for fruiting body yield on composted cotton ginning mill waste substrate under indoor conditions. Results: In successive yield evaluation trials (one preliminary and three full scale), hybrid strains VvH-11, VvH-13 and VvH-18 gave fruiting body yield higher than the two parents. Out of these hybrids, the yield distribution at different height (selves) in growing room was consistent in hybrid VvH-13 in trial-2, while rest two hybrids and parents showed significantly lower yield in lowest self (30 cm above floor level). The fruiting body weight also showed similar trend. The fruiting bodies from the highest yielding hybrid VvH-11 exhibited highest level of crude fiber (2.07%) and ash (10.95%) contents, while the hybrid VvH-13 giving consistent yield across growing room exhibited higher level of crude fiber (2.00 %), vitamin C (52.35 mg 100 g-1) and vitamin D (1434.7 µg 100 g-1). The third hybrid VvH-18 was superior in crude fiber (1.74%), manganese (22.46 ppm), selenium (0.26 ppm) and vitamin C (43.13 mg 100 g-1). All three hybrids exhibited lower fat (1.54 to 1.86 %) content compared to parents (2.54 to 2.59 %). Interpretation: High yielding hybrid strains can be developed in paddy straw mushroom (V. volvacea) through mating of slow growing SSIs, and their screening involving downward mycelial growth on paddy straw and repetitive grow out trials.

Development of a Molecular Marker for Fruiting Body Pattern in Auricularia auricula-judae

The fruiting body pattern is an important agronomic trait of the edible fungus Auricularia auricula-judae, and an important breeding target. There are two types of fruiting body pattern: the cluster type and the chrysanthemum type. We identified the fruiting body pattern of 26 test strains, and then constructed two different near-isogenic pools. Then, we developed sequence characterized amplified region (SCAR) molecular markers associated with the fruiting body pattern based on sequence-related amplified polymorphism (SRAP) markers. Ten different bands (189–522 bp) were amplified using 153 pairs of SRAP primers. The SCAR marker “SCL-18” consisted of a single 522-bp band amplified from the cluster-type strains, but not the chrysanthemum strains. This SCAR marker was closely associated with the cluster-type fruiting body trait of A. auricula-judae. These results lay the foundation for further research to locate and clone genes controlling the fruiting body pattern of A. auricula-judae.

Proteomic Analysis of Coprinopsis cinerea under Conditions of Horizontal and Perpendicular Gravity

Coprinopsis cinerea was employed to investigate the fungal response to gravity. Mycelium growth revealed a consistent growth pattern, irrespective of the direction of gravity (i.e., horizontal vs. perpendicular). However, the fruiting body grew in the direction opposite to that of gravity once the primordia had formed. For the proteomic analysis, only curved-stem samples were used. Fifty-one proteins were identified and classified into 13 groups according to function. The major functional groups were hydrolases and transferases (16%), signal transduction (15%), oxidoreductases and isomerases (11%), carbohydrate metabolism (9%), and transport (5%). To the best of our knowledge, this is the first report on a proteomic approach to evaluate the molecular response of C. cinerea to gravity.

Genet Variation of Ectomycorrhizal Suillus granulatus Fruiting Bodies in Pinus strobus Stands

The genets of Suillus granulatus in a Pinus strobus stand (13 m × 60 m) were identified using random amplified polymorphic DNA molecular markers and the DNA of mushrooms that fruited for two years, and variations in genet size and distribution were analyzed. From a total of 116 mushrooms, 73 genets were identified and were grouped into three locations. The genets of mushrooms in close proximity differed from each other. The genet sizes varied at any of the three locations. The lengths of the identified genets in the pine stand ranged from 0.09 to 2.90 m. The average number of mushrooms per genet was 1.2 to 2.3, and the percentage of genets that were represented by a single mushroom was 44% to 94%. This variation in the genets of mushrooms in close proximity suggests that the ectomycorrhizal mycelial bodies of S. granulatus propagated sexually by fusing haploid spores derived from the mushrooms gills with below-ground mycelia. Therefore, it is necessary further to investigate the formation of new genets through spores in ectomycorrhizal fungal colonies.

Identification of the Genes Involved in the Fruiting Body Production and Cordycepin Formation of Cordyceps militaris Fungus

A mutant library of Cordyceps militaris was constructed by improved Agrobacterium tumefaciens-mediated transformation and screened for degradation features. Six mutants with altered characters in in vitro and in vivo fruiting body production, and cordycepin formation were found to contain a single copy T-DNA. T-DNA flanking sequences of these mutants were identified by thermal asymmetric interlaced-PCR approach. ATP-dependent helicase, cytochrome oxidase subunit I and ubiquitin-like activating enzyme were involved in in vitro fruiting body production, serine/threonine phosphatase involved in in vivo fruiting body production, while glucose-methanol-choline oxidoreductase and telomerase reverse transcriptase involved in cordycepin formation. These genes were analyzed by bioinformatics methods, and their molecular function and biology process were speculated by Gene Ontology (GO) analysis. The results provided useful information for the control of culture degeneration in commercial production of C. militaris.

Entomopathogenicity of Simplicillium lanosoniveum Isolated in Korea

Fruiting bodies similar to those of the ascomycete fungi Podostroma cornu-damae and Cordyceps militaris were collected from Mt. Seunghak in Busan, Korea on August 21, 2012. The fruiting bodies were cylindrical, with tapered ends and golden red in color. The fruiting bodies contained abundant conidiophores bearing single-celled conidia, but no perithecia or asci. Pure culture of the fungal isolates was obtained through single-spore isolation. Analyses of morphological characteristics, including conidia shape, and phylogenetic traits, using internal transcribed spacer sequences, showed that these isolates belonged to the species Simplicillium lanosoniveum. Although this fungal species is known to be mycoparasitic, the isolates obtained in this study were unable to infect fungi. However, silkworms (Bombyx mori) inoculated with the fungal isolates died during the larval or pupal stages, as has been shown for the strongly entomopathogenic fungus Beauveria bassiana. This study is the first report of the entomopathogenicity of S. lanosoniveum and indicates its potential for use in biological control of insects.

Effect of Agaricus brasiliensis and Lentinula edodes mushrooms on the infection of passionflower with Cowpea aphid-borne mosaic virus

The objective of the present study was to evaluate the protection of passion fruit plants against CABMV by using preparations from Agaricus brasiliensis and Lentinula edodes mushrooms. In experiments carried out in the greenhouse, the fruiting body extracts from some of the isolates of both mushrooms significantly reduced CABMV incidence in passion fruit plants. This protective effect occurred when the plant leaves, pre-treated with extracts, were later inoculated mechanically with the virus. However, the extracts did not protect the plants in experiments involving CABMV transmission by aphid vectors. An inhibitory effect of mushroom extracts on the virus particles was also demonstrated on Chenopodium quinoa, a CABMV local lesion host, by inoculating the plants with a mixture of extracts and virus suspension. Still in C. quinoa, the mushroom extracts from some isolates induced systemic resistance against the virus. These results showed that aqueous extracts from A. brasiliensis and L. edodes fruiting bodies had CABMV infectivity inhibitors, but that was not enough to control the viral disease on passion fruit plants at all, considering they were infected through a vector.

O endurecimento dos frutos do maracujazeiro, causado pelo Cowpea aphid-borne mosaic virus (CABMV), é um dos problemas mais sérios que atingem a cultura. Tentativas de se obter plantas resistentes ao vírus ou estirpes fracas premunizantes não apresentaram sucesso até o momento. O objetivo do presente estudo foi o de avaliar a proteção das plantas de maracujá contra o CABMV, utilizando preparações dos cogumelos Lentinula edodes e Agaricus blazei, através da indução de resistência. Em experimentos conduzidos no interior de casa de vegetação, os extratos de basidiocarpos de ambos os cogumelos reduziram significativamente a incidência da virose em plantas de maracujá que tiveram as folhas pré-tratadas com esses extratos e que foram posteriormente inoculadas mecanicamente com o CABMV. No entanto, os extratos não protegeram as plantas em experimentos envolvendo a transmissão do CABMV pelo afídeo-vetor. O efeito inibidor dos extratos foi confirmado inoculando-se Chenopodium quinoa com uma mistura de extratos e suspensão viral. Ainda em C. quinoa, um hospedeiro de lesão local do CABMV, os extratos de alguns isolados dos cogumelos induziram resistência sistêmica contra o vírus. Os resultados mostram que os extratos aquosos de basidiocarpos de L. edodes e A. blazei contêm substâncias inibidoras da infectividade do CABMV, mas isso não é o suficiente para o controle pleno da virose em plantas de maracujá, considerando que elas são infectadas através de um vetor.

Optimum Conditions for Artificial Fruiting Body Formation of Cordyceps cardinalis

Stromatal fruiting bodies of Cordyceps cardinalis were successfully produced in cereals. Brown rice, German millet and standard millet produced the longest-length of stromata, followed by Chinese pearl barley, Indian millet, black rice and standard barley. Oatmeal produced the shortest-length of fruiting bodies. Supplementation of pupa and larva to the grains resulted in a slightly enhanced production of fruiting bodies; pupa showing better production than larva. 50~60 g of brown rice and 10~20 g of pupa mixed with 50~60 mL of water in 1,000 mL polypropylene (PP) bottle was found to be optimum for fruiting body production. Liquid inoculation of 15~20 mL per PP bottle produced best fruiting bodies. The optimal temperature for the formation of fruiting bodies was 25degrees C, under conditions of continuous light. Few fruiting bodies were produced under the condition of complete darkness, and the fresh weight was considerable low, compared to that of light condition.

Spore Dispersion of Tricholoma matsutake at a Pinus densiflora Stand in Korea

The spore of Tricholoma matsutake is considered to be the starting point of the mushroom growth cycle, but the mechanism of mycelial development from the spore stage is not yet clarified. In this study, we tried to measure how far the spores of T. matsutake disperse from a fruiting body located at a Pinus densiflora stand in Korea. We established 16 slide glasses coated with glycerin near a fruiting body in four directions separated by four different distance intervals within a mushroom productive stand after removing all other fruiting bodies from three plots. The number of dispersed spores increased with time from the first day (475 spores/cm2) to the fourth day (836 spores/cm2) after the pileus opened. The number of spores dispersed downward was about 1.5 times greater than that dispersed toward the ridge. The number of dispersed spores decreased exponentially as the distance from each fruiting body increased. More than 95% of the spores dropped within a meter from the fruiting body, with 75% dropping within 0.5 m. Even so, the number of spores dispersed over 5 m from the fruiting body was more than 50 million when considering the total number of spores produced by a fruiting body is about 5 billion.

Occurrence of Dumontinia tuberosa in Korea

Thirty-seven single spore isolates were obtained from specimens of ascomycetous fruiting bodies collected from Mt. Suri, Anyang in Korea. The fungal specimens and isolates were identified as Dumontinia tuberosa based on their morphological and cultural characteristics. This is the first record of this fungus occurring in Korea.

Isolation of Genes Specifically Expressed in Different Developmental Stages of Pleurotus ostreatus Using Macroarray Analysis

The oyster mushroom (Pleurotus ostreatus) is one of the most important edible mushrooms worldwide. The mechanism of P. ostreatus fruiting body development has been of interest both for the basic understanding of the phenotypic change of the mycelium-fruiting body and to improve breeding of the mushrooms. Based on our previous publication of P. ostreatus expressed sequence tag database, 1,528 unigene clones were used in macroarray analysis of mycelium, fruiting body and basidiospore developmental stages of P. ostreatus. Gene expression profile databases generated by evaluating expression levels showed that 33, 10, and 94 genes were abundantly expressed in mycelium, fruiting body and basidiospore developmental stages, respectively. Among them, the genes specifically expressed in the fruiting body stage were further analyzed by reverse transcription-polymerase chain reaction and Northern blot to investigate temporal and spatial expression patterns. These results provide useful information for future studies of edible mushroom development.

Cultural Characteristics and Log-Mediated Cultivation of the Medicinal Mushroom, Phellinus linteus

The optimal conditions for mycelial growth of Phellinus linteus ATCC 26710 were determined to be a log length of 20 cm, temperature of 30degrees C and pH of 6.0. Mycelial growth was excellent on the mushroom complete medium, and was optimal when sucrose, mannose and glucose were supplied as carbon sources. Potassium nitrate and sodium nitrate as nitrogen sources supported good mycelial growth. To evaluate P. linteus mycelial colonization on logs, sterilized short log inoculation, drilling inoculation and log-end sandwich inoculation techniques were used. Only sterilized short log inoculation produced good mycelial colonization. Initial mycelial growth and full mycelial colonization were best on 20 cm logs having 42% moisture content. The initial mycelial growth of P. linteus was accelerated over 12 hr of sterilization. Basidiocarp formation was optimal using a burying method of logs after 5~6 months, and fruiting body formation was superior in cultivation house conditions of 31~35degrees C and in excess of 96% relative humidity.

Effect of Preservation Periods and Subcultures on Fruiting Body Formation of Cordyceps militaris In Vitro

Effects of various preservation periods and subcultures on fruiting body formation of Cordyceps militaris were investigated using EFCC C-10995 single ascospore strains. Fruiting body formation by original strains was profuse when preserved at 4degrees C for 5~6 months. Fruiting from subcultures was stable till second to sixth subcultures, after which it decreased sharply. The more the colony color of subcultures changed, the less the fruiting bodies formed. Liquid inoculum preparation of single ascospore strains in the same or separate broths did not affect fruiting body formation. Similarly, two strains C-10995-3 and C-10995-6 in different numbers during liquid inoculum preparation produced similar fruiting bodies.

Cultural Characteristics of A Medicinal Mushroom, Phellinus linteus

For comparison of mycelial colonization of Phellinus linteus on logs, several techniques of inoculation were tested; sterilized short log inoculation, drilling inoculation and log-end sandwich inoculation. The mycelial colonization of P. linteus on logs was good in the treatment of sterilized short log inoculation, but poor in the traditional methods such as drilling inoculation and log-end sandwich inoculation. The initial mycelial growth and the full mycelial colonization of P. linteus in logs were the best in case of 20 cm logs under the condition of 42% moisture content. Also, the initial mycelial growth of P. linteus was accelerated over 12 hours of sterilization. Burying method of logs after 5~6 months of incubation was the best for formation of basidiocarps of P. linteus. The formation of fruiting body of P. linteus was quite good in the cultivation house at 31~35degrees C and over 96% of relative humidity.

Cytotoxicity of Paecilomyces tenuipes Against Human Carcinoma Cells, HepG2 and MCF-7 In Vitro

The methanolic extract of fruiting body of Paecilomyces tenuipes DGUM 32001 showed significant cytotoxicity against human cancer cells: HepG2 and MCF-7. The methanolic extract was further fractionated with organic solvents such as chloroform and ethyl acetate in that order. Among the fractions tested, the ethyl acetate fraction showed the highest cytotoxicity against the carcinoma tested. The IC50 values of ethyl acetate fraction against HepG and MCF-7 were 40 and 9.6 microg/ml, respectively.

Structure identification of triterpenes from fruiting bodies of Ganoderma lucidum by NMR spectra and X-ray diffraction analysis / 中草药

Object To study the chemical constituents from fruiting bodies of Ganoderma lucidum (Leyss ex Fr.) Karst Methods Individual constituent was isolated and repeatedly purified on silica gel column and structurally elucidated by the NMR spectra and X ray diffraction analysis Results Four compounds were obtained from the alcohol extract and identified as 3? oxo formyl 7?, 12? dihydroxy 4, 4, 14? trimethyl 5? chol 11, 15 dioxo 8 en (E) 24 oic acid (Ⅵ); 3? oxo formyl 7?, 12? dihydroxy 5? lanost 11, 15, 23 trioxo 8 en (E) 26 oic acid (Ⅶ); ergosterol (Ⅷ) and ergosta 7, 22 dien 3? ol (Ⅸ), respectively Conclusion Compound Ⅵ and Ⅶ are new triterpenes