• Mycobiology
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• v.35 no.2
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• pp.54-61
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• 2007

Two isolates of Tricholoma matsutake T-008 and T-034, preserved in Entomopathogenic Fungal Culture Collection (EFCC) of Korea, were used in the present study. The isolates had 100% Bootstrap homology with Tricholoma matsutake U62964 and T. matsutake AB188557 and AF309538 preserved in Gene Bank of NCBI. Mycelial growth of T. matsutake was highest in TMM and MYA at $25^{\circ}C$. The highest dry wt. of mycelium was obtained after 65 days of culture, when 6 mycelial discs were inoculated in 100 ml of broth in 250 ml shaking flask. Mycelial mats were observed in clumped condition at the inoculation sites of pine forest after two weeks of inoculation. After 5 months of inoculation, mycelia mats were observed growing inside soil and walls of a few inoculation sites, while mycelial mats growth up to $5{\sim}8$ cm were observed in the roots of pine tree after 6 months. The survival rate of the inoculum was about 40% of the total inoculation sites. The survival rate was found below 20% when the mycelium was inoculated in the summer. The reasons for low survival rates of the mycelium were mainly due to dry season and the soil-borne small animals such as earthworm and mole. After one year of inoculation, no external difference was observed between the artificially inoculated mycelia and the naturally existing mycelia of T. matsutake. The present study showed that fruiting bodies of T. matsutake could be produced by artificial inoculation under the appropriate environmental conditions.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.355-358
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• 2001

Selection of optima] nutrient sources and cultural methods for liquid spawn culture of Flammulina velutipes were carried out. The optimal temperature and pH for mycelial growth of F. velutipes were $20^{\circ}C$ and 6.0 to 7.5. respectively. In the 250ml ${Delta}$-flask culture. the amount of inoculum and culture period for the optimal mycelial growth of F. velutipes were 3 mycelial disks(diametcr 6mm) and 6 days, respectively. For the mass production of submerged culture. the optimal inoculum amount and aeration rate of F. velutipes were 5%(inoculum vol/medium vol.) and l.0vvm(vol of air/vol. of medium/min), respectively.

• Journal of Microbiology and Biotechnology
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• v.14 no.5
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• pp.944-951
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• 2004

The influences of inoculum size, pH, and medium composition on mycelial growth and exopolysaccharides (EPS) production were investigated in shake flasks and in a bioreactor. The optimum inoculum size for both mycelial growth and EPS production was identified to be 10% (v/v) in shake flask cultures. The optimal initial pH for mycelial growth and EPS production in shake flask cultures were found to be 5.0 and 7.0, respectively. However, the optimal pH was 5.0 for both mycelial growth and EPS production in bioreactor cultures where the pH was regulated. The optimal mass ratio of the two major carbon sources, glucose to dextrin, was 1:4. The optimal mass ratio of the two major nitrogen sources, yeast extract to soy tone peptone, was 2:1. When 500 mg $1^{-1}$ of $MnSO_4-5H_2O$ was added to the bioreactor culture, both mycelial growth and EPS production were enhanced by approximately 10%. Under the optimized conditions, a mycelial biomass of 9.85 g $1^{-1}$ and an EPS concentration of 4.92 g $1^{-1}$ were obtained in 4 days.

• Korean Journal of Agricultural Science
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• v.44 no.3
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• pp.339-347
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• 2017

Sclerotium rolfsii causing southern blight on numerous vegetable and fruit crops was isolated from stems of chili peppers showing wilting symptoms. The pathogen was identified by morphological observation and DNA sequencing analysis of ITS region. To select an effective fungicide for control of southern blight, we investigated the inhibition efficacy of thirty fungicides included in nine groups of fungicides with different mechanisms of action. A fungal growth inhibition assay was conducted through an agar dilution method by using mycelial discs and sclerotia of the pathogen as inoculum, respectively. When mycelial discs were used as an inoculum, several fungicides showed good inhibitory activity against the mycelial growth of S. rolfsii 12-6. All DMI fungicides tested had a good inhibition except for prochloraz which had low inhibitory effect. All strobilurin fungicides tested except for kresoxim-methyl and all SDHI fungicides tested except for boscalid and fluopyram, had a good inhibition. Also, fludioxonil, a protective fungicide and fluazinam had a good inhibitory effect. Interestingly, when sclerotia were used as an inoculum, inhibition efficacy was increased for fluopyram, a SDHI fungicide, and for some protective fungicides such as propineb, chlorothalonil, dithianon, and folpet. All the fungicides selected in this study should be tested in the field for their control activities against stem rot for practical use in chili pepper cultivation.

• The Korean Journal of Mycology
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• v.26 no.3 s.86
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• pp.387-395
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• 1998

Mycelial growth, production of microsclerotia and perithecia, and presence of double-stranded RNA were examined in Calonectria ilicicola isolates from several hosts to detect morphological and/or genetic markers for comparison with levels of virulence. Variability in disease severity, production of microsclerotia and perithecia, and mycelial growth was observed across all isolates. None of 35 isolates of C. ilicicola examined contained detectable levels of double-stranded RNA. Disease severity on soybean cultivars correlated positively with production of both microsclerotia and perithecia. Mycelial growth correlated negatively with production of perithecia. Virulence on the cultivars and production of microsclerotia and perithecia were greater in isolates of C. ilicicola from soybean than those from peanut. These results suggest that the ability of isolates to produce microsclerotia and perithecia is a component of inoculum potential. Perithecia production may serve as a useful marker for characterizing virulence or host specialization in this homothallic fungus.

• Applied Biological Chemistry
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• v.42 no.3
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• pp.188-192
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• 1999

In order to minimize the mycelial pellet formation, one of the critical obstacles during the fermentation processes of filamentous fungi, an investigation was focused on the culture conditions(media and initial inoculum) and additives(soils, surfactants and polyethylene glycol 200) when a high phosphate-dissolving fungus, Penicillium sp. GL-101, was cultured in liquid media. Culturing the strain in PDB, SDB and YPD media, their pellet sizes decreased to the order of YPD > SDB > PDB. And at the high concentrations of the initial inoculum in the range from $1{\times}10^3\;to\;1{\times}10^6$ conidia/ml, the small sizes of pellet were formed in the PDB media. For the initial inoculum between $1{\times}10^7\;and\;1{\times}10^8$ conidia/ml, however, an amorphous pellet or loose aggregate was formed. The addition of soils, zeolite and diatomite, up to 1.0% decreased the pellet sizes to 3/4 and 1/2, respectively, but the pellet was increased to 2.5 times by the addition of bentonite. Surfactants also affected on the size of pellet; the addition of Triton X-100 and Tween 80 up to 1.0% decreased the pellet sizes maximally to 1/10 and 1/4, respectively, while SDS completely inhibited the fungal growth. Among the four additives tsted, polyethylene glycol 200 was the most effectively reduced the pellet sizes to $0.2{\pm}0.1$mm that resulted in about 25- fold reduction compared to the control.

• The Korean Journal of Mycology
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• v.27 no.1 s.88
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• pp.1-9
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• 1999

For the practical using of liquid spawn we carried out selection test of nutrient sources, cultural methods and cultural apparatus for liquid spawn production of oyster mushroom(ASTI 2001, ASTI 2018, ASTI 2072, ASTI 2016, ASTI 2070, ASTI 2180, ASTI 2183, ASTI 2042). The optimal temperature and pH range for mycelial growth of Pleurotus species were $25^{\circ}C\;to\;30^{\circ}C$ and 5.5 to 6.5, respectively. The effect of carbon sources, nitrogen sources and mineral salts on the mycelial growth was studied using petridish culture. Generally, the disaccharides and polysaccharides showed good effect for mycelial growth of Pleurotus species, and the polysaccharides were superior to the other classes of carbon sources for mycelial growth. For the mycelial growth of the 8 oyster mushroom stains, soybean flour was superior to the other kinds of nitrogen sources. On the other hand, addition of mineral salts did not affect, and even poor under certain mineral salts, the mycelial growth of the 8 oyster mushroom stains. The brown sugar selected out the carbon source of the agricultural medium. Also the soybean flour selected out the nitrogen source of agricultural medium. In the medium selection, we selected out agricultural optimum medium composed of brown sugar 3%, soybean flour 0.3%, potassium phosphate 0.05%, magnesium sulfate 0.05%. Under the 250 ml erlenmeyer culture, the effects of such factors as the inoculum rate, the working volume, cultural method and flask shapes on the mycelial growth were examined. The optimal inoculum rate and working volume on the mycelial growth of oyster mushroom was 2 mycelial disk (diameter 6 mm) and 100 ml, respectively. The shake flask culture was enhanced the mycelial growth than at the erlenmeyer flask. Pulp form growth of mycelium in the erlenmeyer flask culture was obtained in the culture with glass rod of length 50 mm, diameter 10 mm.

• Mycobiology
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• v.37 no.2
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• pp.89-93
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• 2009

The effect of fermentation parameters and medium composition on the simultaneous mycelial growth and exo-polymer production from submerged cultures of Ganoderma applanatum was investigated in shake-flask cultures. The optimum initial pH for mycelial growth and exo-polymer production was 5.0 and 6.0, respectively. The optimum temperature was $25^{\circ}C$ and the optimum inoculum content was 3.0% (v/v). The optimal carbon and nitrogen sources were glucose and corn steep powder, respectively. After 12 days fermentation under these conditions, the highest mycelial growth was 18.0 g/l and the highest exo-polymer production was 3.9 g/l.

• Korean Journal Plant Pathology
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• v.14 no.1
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• pp.46-51
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• 1998

A method for inoculum production of Botryosphaerisa dothidea was developed using cucumber disks and cereal media. Disks of cucumber fruits, and cereal media of barley, wheat, and rice seeds were inoculated with mycelial plugs of B. dothidea and incubated at 27$^{\circ}C$. Pycnidia were produced on the surface of cucumber disks and seeds after 5 days of inoculation. When the inoculated barley seeds were immersed in sterilized distilled water for 5 minutes, abundant conidia of B. dothidea were exuded from mature pycnidia. Conidia were held together by mucilage as they were released from an ostiole. Compared with the conventional method for inoculum preparation using agar media, such as potato-dextrose agar and oatmeal agar, this method could minimize the tedious work required for inoculum preparation within a shorter period of time.

• Korean Journal Plant Pathology
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• v.13 no.2
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• pp.105-112
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• 1997

Didymella bryoniae, gummy stem blight fungus of cucurbits, has been known not to produce its pycnidium in vitro without irradiation. Various methods for producing pycnidiospores of the fungus as an inoculum have been used. However, those methods have not been verified in terms of efficiency of the productivity, activity and synchronous maturation of the inoculum. Therefore, a pycnidiospore production method in vitro that is highly reliable and reproducible has to be developed to obtain a large amount of inoculum for screening disease resistant varieties or effective fungicides. Here we standardized a mass-production technique for pycnidiospores of D. bryoniae in vitro by comprehensively finding the optimal conditions such as kinds and thickness of cultural medium, growing temperature, and quality and duration of irradiation as well as examining the activity and pathogenicity of the pycnidiospores reproduced. In brief, mycelial colony on the PDA plate was cultured at 26$^{\circ}C$ for 2 days under the darkness, and then the plate was irradiated under the UV light (12 hr/a day) for 2~3 days at the same temperature(26$^{\circ}C$). Two days after UV irradiation, a great number of pycnidia was simultaneously formed. This plate was subjected to darkness again for 4~5 days to mature pycnidiospores. We could obtain a large amount of inoculum that is synchronously matured in a short period of time through the above procedures.