Mushroom-derived ${\beta}$-glucan, a polysaccharide (GLP) isolated from the mycelium of Ganoderma lucidum, was previously shown to have inhibitory effects against tumor-bearing mice in vivo. We investigated the apoptotic effect of mushroom-derived ${\beta}$-glucan in a sarcoma-180 tumor cell- bearing mice model using an ELISA to determine the levels of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in the mice. The morphology of the tumor cells was assessed with transmission electron microscopy (TEM). GLP was injected into the tumor-bearing mice at a dose (i.p.) of 20 mg/kg for 10 days. After 30 days, the tumor mass from the inguinal region was collected, weighed, and assayed using TEM and a TNF-${\alpha}$ ELISA kit. The tumors that developed in the mice treated with GLP were 71.4% smaller than those in the control group, showing the ability of GLP to inhibit tumor growth. The levels of TNF-${\alpha}$ in the serum of the sarcoma-180 bearing mice were 12 times greater than in the serum of the nonbearing tumor mice. An ultrastructural study demonstrated that the GLP-treated sarcoma-180 tumor cells were condensed, with rearranged chromatin. In addition, the marginated chromatin in nucleus induced the nuclear compartment, and there were many vacuolization in the cell. GLP could be an effective apoptosis-inducing compound in sarcoma-type cancers.
Cultivation characteristics of 12 strains of cauliflower mushroom (Sparassis crispa) collected in Korea were investigated by growing the mushroom on sawdust medium of Larix kaempferi. As cultivation characteristics, incubation period for full growth of mycelium in a cultivation bottle, cultivation time period taken for first harvest, and mushroom color and yield were examined. S. crispa KFRI 723 showed the shortest for incubation period with 59 days while S. crispa KFRI 746 showed the longest with 94 days. The earliest mushroom harvesting was achieved by 29 days from S. crispa KFRI 746 and the latest was by 63 days from S. crispa KFRI 691. The colors of fruit body of the tested strains can be divided into three groups; S. crispa KFRI 700 was white, S. crispa KFRI 747 was yellow brown, and the others were light yellowish. KFRI 700 yielded the most as 163 g from 380 g sawdust media, while KFRI 746 and KFRI 747 were the lowest with 58 g and 35 g, respectively. As results of cultivation characteristics of 12 strains of cauliflower mushroom, we consider that three strains (KFRI 700, 723 and 724) of S. crispa are suitable for sawdust cultivation on L. kaempferi in the aspects of mycelial growth period, harvesting period and mushroom production, respectively.
Lee, Sang Myeong;Kim, Dong Soo;Lee, Kwang-Soo;Lee, Chong-Kyu;Lee, Dong Woon
Weed & Turfgrass Science
/
v.2
no.2
/
pp.202-206
/
2013
Biocontrol potential of an isolate of Helicosporium spp. against Rhizoctonia solani, Fusarium oxysporium and Phytophthora drechsleri was evaluated in vitro and in vivo. A selected biocontrol agent designated as Helicosporium 0635BP strongly inhibited growth and lysed mycelium of Rhizoctonia solani and Fusarium oxysporium on PDA. Autoclaved culture filtrate of the agent also completely inhibited growth of the turfgrass large patch pathogen, R. solani AG2-2 IV at the concentration of 50 ml $L^{-1}$. The pathogen was killed when dipped under the 20% filtrate for four hours or 50% for one hr. In a field trial, plots applied with the crude or times diluted culture filtrate showed 100% control efficacy of the turfgrass large patch as a chemical applied for a comparison. Results indicated that Helicosporium 0635BP is a promising biocontrol agent on control of the turfgrass large patch disease and its culture filtrate contained unknown heat suitable antifungal substance (s). Further studies on mass production, purification and identification of the unknown compound (s) are in progress for practical use.
Kim, Jeong-Ho;Shim, Gyu-Yul;Lee, Hye-Min;Moon, Hyo-Sun;Kim, Young-Ho
Asian Journal of Turfgrass Science
/
v.21
no.2
/
pp.147-154
/
2007
In March of 2004, gray snow mold (Typhula blight) caused by Typhula spp. occurred on perennial ryegrass (Lolium perenne L.) and Kentucky bluegrass (Poo pratensis L.) at MuJu golf courses in Jeonbuk Province. Leaves in the affected areas were matted together and frequently covered with white to grayish mycelia. Sclerotia were formed on the leaf blade, leaf sheath, or crown regions. The fungus isolated from the diseased leaf formed whitish mycelium, clamp connections, and light pink to brown, irregular-shaped small sclerotia of less than 1.4 mm in diameter, which are characteristic to Typhula incarnata. Optimum temperature ranges for mycelial growth were $5^{\circ}C$ to $15^{\circ}C$. The causal organism was confirmed to be T. incarnata as the partial sequence of its ribosomal RNA ITS1 (internal transcribed spacer) region was 91% homologous to those of T. incarnata in GenBank database. Out of the 14 fungicides tested fur antifungal activity in vitro, 10 fungicides including iprodione, tebuconazole, polyoxin D, flutolanil, hexaconazole, tolclofos-methyl, fosetyl-Al, mepronil, pencycuron+tebuconazole, and fenarimol completely inhibited fungal growth at their recommended concentrations. In the field test, these fungicides and others such as thifluzamide and thiram effectively controlled the gray snow mold of turfgrass with some variable degrees of control efficacies.
Recalcitrant 4-t-Octylphenol used as a surfactant was subjected to the biodegradation with wood rot fungi, Basidioradulum molare and Schizopora paradoxa. Two fungi were grown in the culture medium containing various concentrations of 4-t-Octylphenol in order to investigate their resistance against 4-t-octylphenol Schizopora paradoxa was reached to the full growth within 14 incubation days in the concentration of more than 200 ppm of 4-t-Octylphenol, while Basidioradulum molare showed the inhibitory mycelium growth as the concentration was increased Schizopora paradoxa and Basidioradulum molare biodegraded 95% and 36% of initial concentration of 4-t-Octylphenol at first incubation day, respectively. However, the biodegradation capability reached to more than 95% after 3 incubation days. During the biodegradation of 4-t-Octylphenol, the activity of manganese dependent peroxidase was induced by the addition of 4-t-Octylphenol in the culture medium of Schizopora paradoxa, but that of laccase was maximal before the addition. The reduction of estrogenecity was assayed by MCF-7 cell proliferation test and measurement of pS2 mRNA expression. The level of pS2 mRNA was decreased down to the level of baseline at first incubation day. Also, estrogenecity of 4-t-Ocrylphenol completely disappeared after treatment with supernatant by Schizopora paradoxa and Basidioradulum molare from first incubation day of culture down to the levels of vehicle.
BACKGROUND: In order to select a fungicide that can effectively control anthracnose disease in Japanese plum fruit, mycelial growth inhibition effect and spore germination inhibition effect of six fungicides were tested in vitro against six isolates of Colletotrichum acutatum and five isolates of C. gloeosporioides that were isolated from diseased Japanese plum fruit. METHODS AND RESULTS: Inhibitory effects of fungicides on mycelial growth were investigated after inoculating each isolate on potato dextrose agar amended with four discriminatory concentrations of each fungicide for 7 days at $25^{\circ}C$. For spore germination inhibitory effect, each isolate of the Colletotrichum spp. was cultured in potato dextrose agar for 7-14 days at $25^{\circ}C$. After adjusting the concentration of spores of each isolate to $1{\times}10^6mL^{-1}$ by diluting with 0.025% PDB, the spore suspension was mixed with each fungicide (1:4, v/v), and $60{\mu}L$ aliquots were dispensed to sterile hole slide glass. Hole slide glasses were placed in a humidified box and incubated for 15 hours at $25^{\circ}C$. Then, spore germination was observed under an optical microscope. At recommended concentration of fungicide prochloraz manganese showed the highest mycelial growth inhibitory effect and dithianon showed the lowest mycelial growth inhibition. The $EC_{50}$ values for the inhibition of spore germination by dithianon and pyraclostrobin were $0.069-0.126{\mu}g/mL$ and $0.37-1.59{\mu}g/mL$, respectively. Although benomyl, prochloraz manganese, azoxystrobin, and tebuconazole did not inhibit the spore germination, they appeared to restrain mycelial growth by abnormal growth of germ tube and mycelium after germination. CONCLUSION: Dithianon seemed to have preventive effect. Prochloraz manganese, azoxystrobin, and tebuconazole were likely to have control effect. Pyraclostrobin is considered to have both preventive and control effect against anthracnose disease of Japanese plum fruit.
Objective of this study was to select ectomycorrhizal fungi for black spruce(Picea mariana) inoculation to overcome the growth inhibitory effects of Kalmia angustifolia. Nineteen isolates representing 11 species of ectomycorrhizal fungi were tested for their abilities to grow and form mycorrhizae with black spruce seedlings in the presence of water leachate of leaves of Kalmia. Mycelium growth of 9 isolates were inhibited by the leaf leachate. Colony diameter and biomass of the other 10 isolates were either increased or unaffected under the same conditions. Acidic pH of the culture medium(pH 3 and 4) inhibited some of the fungi, but a combination of acidic pH and the leaf leachate was more inhibitory. Thirteen isolates were able to form ectomycorrhizae with black spruce in presence of 25% leaf leachate in pure culture. Four isolates, Paxillus involutus(NF4), Cenococcum geophilum(GB12), Laccaria laccata(GB23), and E-strain(GB45) formed mycorrhizae more successfully than the others in presence of up to 50% Kalmia leaf leachate. Black spruce seedlings pre-inoculated with these fungi were grown with Kalmia leaf leachate and live Kalmia plants during a four month greenhouse experiment. Abundant mycorrhizae(77-91% of root tips) were developed on seedlings pre-inoculated with P. involutus, L. laccata and E-strain but relatively poor mycorrhization(32% of root tips) resulted with C. geophidum. Over 90% of the short root mycorrhizae were attributed to the inoculated fungi although indigenous mycorrhizae also occurred on most seedlings. Persistence of the mycorrhizae was not affected by living Kalmia plants. Over 80% of the mycorrhizae on seedlings inoculated with P. involutus, L. laccata and E-strain and 53% of the mycorrhizae on seedlings inoculated with C. geophilum were attributable to the inoculant fungi. Control seedlings formed about 45% ectomycorrhizal short roots with indigenous fungi. The L. laccata and C. geophilum inoculated seedlings exhibited enhanced mycorrhizae formation in presence of Kalmia leaf leachate. Mycorrhizae formation with inoculant fungi was 4-15% lower at pH 4 than at pH 5, with the greatest inhibition occurring for L. laccata. Seedlings inoculated with P. involutus had the greatest shoot and root growth followed by L. laccata and E-strain inoculated seedlings. The P. involutus and L. laccata inoculated seedlings were significantly taller with more shoot dry biomass than the uninoculated(control) seedlings. E-strain inoculated seedlings had significantly higher shoot dry biomass and significantly lower number of first order lateral roots compared to the control but other growth parameters such as height, root dry weight and number of short root tips were not significantly different from the control. Seedlings inoculated with C. geophilum were not significantly different from the uninoculated seedlings in any of the growth parameters except for the number of first artier lateral roots which was significantly less than the control seedlings.
Sung, Jae-Mo;Yang, Kun-Joo;Kim, Soo-Ho;Harrington, Tom
The Korean Journal of Mycology
/
v.25
no.1
s.80
/
pp.46-67
/
1997
From 1985 to 1993, we collected 20 isolates throughout Kangwon and obtained 6 isolates from other sources. A. mellea formed rhizomorph actively, and some of A. osroyae were poor in the formation of rhizomorph and some without formation of rhizomorph. A. tabescens was active in the growth of aerial mycelium and poor in the development of rhizomorph. In A. gallica, the mycelium development among the isolates were variable greatly, and especially in isolate A8(KNU-250), the mycelial development was similar to that of A. osroyae, but A8(KNU-250) showed the feature of A. gallica to change medium into brown color. In PCR-RFLP analysis of the IGS region in rDNA, the homology between each isolate in the A. mellea and A. ostoyae showed 100% homology. A. tabescens showed $0.919{\sim}0.974$ homology, and A. gallica showed $0.619{\sim}1.000$ homology. A19 and A12 showed 100% homology as the same group, but compared with other subgroups they showed less than 10% homology as $0.051{\sim}0.108$ value. In RAPD analyses, the isolates of A. mellea showed high homology among themselves as $0.983{\sim}1.000$, and A. ostoyae also showed high similarity. The homology between isolates of A. tabescens showed $0.594{\sim}0.953$ value because A. gallica showed $0.280{\sim}0.733$ value, and the variations between isolates were greater than other species. Especially, A19 and A22 were identified as new novel group which were remoted from other groups, and the homology between these two isolates showed 0.921 value, and the genetic similarity between these groups and other 4 groups showed less than 7% as $0.012{\sim}0.069$ value. Of 5 species identified in this study, 4 species were identified as A. mellea, A. ostoyae, A. tabescens, and A. gallica that were already reported ones and 1 species was suggested as a new specie in Korea.
The key for cultivating Lentinula edodes in sawdust bags with an appropriate strain and medium is to encourage the mushroom growth, while discouraging contaminating fungi by controlling environment, especially temperature and relative humidity (RH). To investigate the daily and seasonal fluctuation of temperature and RH in two L. edodes cultivation sheds types, HOBO data loggers was set and the collected data were analyzed. In a Taiwan type L. edodes cultivation shed, temperature and humidity changes were divided into five characteristic periods: mycelium growing winter, mushroom fruiting spring, mushroom fruiting early summer, mushroom nonfruiting summer and mushroom fruiting autumn. First, the mycelium growing winter was December to early March with daily mean temperature of $-1{\sim}8^{\circ}C$. Second, mushroom fruiting spring was mid March to late May with daily mean temperature of $8{\sim}21^{\circ}C$ and day-night temperature difference of $15^{\circ}C$. Third, the Mushroom fruiting early summer was early June to early July with 17 to $25^{\circ}C$. Fourth, nonfruiting summer was mid July to mid August with daily mean temperature of $25{\sim}28^{\circ}C$. Lastly, mushroom fruiting autumn was late August to October with daily mean temperature of $10{\sim}23^{\circ}C$ and with cyclic temperature change by $7^{\circ}C$ decrease and 5 increase every 5 to 7 days. In a Chinese type shed, temperature ranged $-1.9{\sim}5.0^{\circ}C$ during winter and $15{\sim}32^{\circ}C$ during June to October. Temperature and relative humidity changed $12{\sim}30^{\circ}C$ and 40~100%, respectively, depending on 0~150 cm shelf heights of by positions in the shed. In conclusion, to grow L. edodes but to discourage contaminating fungi, that is, not to be too high in temperature and RH, the growers changed temperature and RH by adjusting shading, aeration and insulation in the shed.
Journal of the Korean Society of Food Science and Nutrition
/
v.43
no.3
/
pp.411-418
/
2014
Three mushroom mycelia, Ganoderma lucidum, Hericium erinaceum, and Phellinus linteus, were separately diluted with the natural culture media Acanthopanax senticosus. Solid-state fermentation was used to produce three different A. senticosus-fermented mushroom mycelium groups: G. lucidum mycelia, H. erinaceum mycelia, and P. linteus mycelia. The resulting mycelia were analyzed to assess their efficacies as health functional foods. Optimized fermentation conditions were determined by considering the density and growth speed of mycelia in each A. senticosus-fermented mushroom mycelium group. The cultured mushroom mycelia under the optimized conditions were extracted using water and 70% ethanol. Extraction was followed by filtration, concentration and freeze-drying to produce extract powder of A. senticosus-fermented mushroom mycelia: Water extracts (FM-5111, FM-5121, and FM-5131) and 70% ethanol extracts (FM-5112, FM-5122, and FM-5132). Analysis of extract powder of A. senticosus-fermented mushroom mycelia was performed using the maker compounds eleutheroside B and eleutheroside E. Analysis of ${\beta}$-glucan contents was performed by enzymatic procedures.
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