• Mycobiology
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• v.48 no.4
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• pp.313-320
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• 2020

In Pleurotus sp., green mold, which is considered a major epidemic, is caused by several Trichoderma species. To develop a rapid molecular marker specific for Trichoderma spp. that potentially cause green mold, eleven Trichoderma species were collected from mushroom farms and the Korean Agricultural Culture Collection (KACC). A dominant fungal isolate from a green mold-infected substrate was identified as Trichoderma pleuroticola based on the sequences of its internal transcribed spacer (ITS) and translation elongation factor 1-α (tef1) genes. In artificial inoculation tests, all Trichoderma spp., including T. atroviride, T. cf. virens, T. citrinoviride, T. harzianum, T. koningii, T. longibrachiatum, T. pleurotum, and T. pleuroticola, showed pathogenicity to some extent, and the observed symptoms were soaked mycelia with a red-brown pigment and retarded mycelium regeneration. A molecular marker was developed for the rapid detection of wide range of Trichoderma spp. based on the DNA sequence alignment of the ITS1 and ITS2 regions of Trichoderma spp. The developed primer set detected only Trichoderma spp., and no cross reactivity with edible mushrooms was observed. The detection limits for the PCR assay of T. harzianum (KACC40558), T. pleurotum (KACC44537), and T. pleuroticola (CAF-TP3) were found to be 500, 50, and 5 fg, respectively, and the detection limit for the pathogen-to-host ratio was approximately 1:10,000 (wt/wt).

• Korean Journal of Microbiology
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• v.22 no.4
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• pp.207-214
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• 1984

Improved methods for the isolation and purification of conidial protoplast were investigated and conidial proplast from auxotrophic mutants were fused. The reaction time for isolation of protoplasts from the swollen condiospores preincubated in liquid minimal medium supplimented with 2-deoxy-D-glucose was shorten by reaction with mixture of 2% driselase and 2% ${\beta}-glucuronidase$ (1:1). The conidial protoplast could be highly purified by using 5% Ficoll 400 as a centrifugation medium. Nucleus of the conidial protoplast was stained with Giemsa stain and the conidial protoplast had one nucleus. It was also confirmed that the conidial protoplast was true protoplast with no cell wall remnant at the outside of plasma menbrane. Fusion frequencies of the conidial protoplast from auxotrophic mutants ranged from $3.4{\times}10^{-1}\;to\;4.9{\times}10^{-1}$. These values were higher than those of mycelial protoplast by a factor of 5 to 28.

• Mycobiology
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• v.33 no.2
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• pp.90-96
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• 2005

Sixty two out of the sixty four species of fungal isolates tested could produce both $exo-{\beta}1,4-gluconase\;(C_1)$ and $endo-{\beta}1,4-gluconase\;(C_x)$ on pure cellulose and rice straw as carbon source in Czapek's medium. Fifty-eight and fifteen species were able to grow at $25^{\circ}C$ and at $45^{\circ}C$, respectively. Eleven species could grow at both $25^{\circ}C$ and $45^{\circ}C$ while, four species appeared only at $45^{\circ}C$. The most cellulolytic species at $25^{\circ}C$ was Trichoderma koningii producing 1.164 $C_1$ (mg glucose/1 ml culture filtrate/1 hr) and 2.690 $C_x$ on pure cellulose, and 0.889 $C_1$, and 1.810 $C_x$ on rice straw, respectively. At $45^{\circ}C$, the most active thermotolerant species were Aspergillus terreus, followed by A. fumigatus. Talaromyces thermophilus was the highest active thermophilic species followed by Malbranchea sulfurea. Most of these species were also active in fermentation of rice straw at 25 and $45^{\circ}C$ (P<0.05). The most active ones were T. koningii, A. ochraceus and A. terreus, which produced 201.5, 193.1 and 188.1 mg crude protein/g dry straw, respectively.

• Mycobiology
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• v.34 no.1
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• pp.14-21
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• 2006

Crude protein (CP) content of mechanically ground rice straw into small particles by an electric grinder and reducing value (RV) and soluble protein (SP) in the culture filtrate were lower than that of the chopped straw into $5{\sim}6\;cm$ lengths when both ground and chopped straws were fermented with Aspergillus ochraceus, A. terreus or Trichoderma koningii, at steady conditions. The reduction rate of RV, SP and CP was 22.2, 2.4, 7.3%; 9.1, 4.9, 8.5% or 0.0, 0.0, 3.6% for the three fungi, respectively. Chemical pretreatment of straw by soaking in $NH_{4}OH$ for a day caused significant increase in CP of the fermented straw than the other alkali and acidic pretreatments. Gamma irradiation pretreatment of dry and wet straw with water, specially at higher doses, 100, 200 or 500 kGy, caused significant increase in RV and SP as CP in the fermented straw by any of these fungi. Chemical-physical combination pretreatment of rice straw reduced the applied dose of gamma irradiation required for increasing fermentable ability of fungi from 500 kGy to 10 kGy with approximately the same results. Significant increases in RV and SP of fermented straw generally occurred as the dose of gamma irradiation for pretreated straw, which combined with $NH_{4}OH$, gradually rose. Whereas, the increase percentage in CP of fermented straw that was pretreated by $NH_{4}OH-10\;kGy$ was 12.4%, 15.4% or 8.6% for A. ochraceus, A. terreus or T. koningii, respectively.

• Korean Journal of Environmental Biology
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• v.20 no.2
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• pp.130-135
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• 2002

Species of Genus Trichoderma are commercially applied as biological control agents against fungal Pathogens. A powerful biocontrol agent, Trichoderma sp. SJG-99721 was isolated from 305 isolates by morphological characters, chitinase activities and antifungal activities against Phytophthora capsiei. The isolate was identified as Trichoderma harzianum from various features such as growth rate at $27^\circ{C}$, significant growth ratio of $27^\circ{C}$ to $17^\circ{C}$, amount of aerial mycelium, types of branching: system, and disposition patterns of phialide and phialospore. Trichoderma harzianum SJG-99721 have been shown to act as a powerful biological agent against fungal phytopathogens; Botrytis cinerea, Rhizoctonia solani, Phytophthora cryptogea, Phytophthora capsiei, Sclerotinia sclerotiorum, Mycoshaerella melonis, Alternaria sotani, Fusarium oxysporum, Collectotrichum gloesporioodes, Alternaria alternata, Phythium ultimum, Phytophthora drechsleri, Pyricularia grisea.

• The Korean Journal of Mycology
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• v.36 no.1
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• pp.9-15
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• 2008

For the detection of contaminated liquid spawn, we selected suitable medium, indicator and developed method of diagnosis. The growth of pathogenic bacteria, Pseudomonas sp., and fungi, Trichoderma sp., in YPL media was better than in PDA and NA. In addition, the changes of color and absorbance of media were obviously showed when contaminated liquid spawn by pathogenic bacteria and fungi was incubated on YPL including phenol red for 48 hour at $25^{\circ}C$. The color of YPLP after incubating of infected liquid spawn by Pseudomonas sp. and Trichoderma sp. were changed from orange to red and to scarlet, respectively. Whereas, the color of YPLP after incubation of only Pleurotus ostreatus indicated yellow at liquid spawn. Therefore, it is possible to easily distinguish contaminated liquid spawn by color of change in YPLP.

• 보존과학연구
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• s.19
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• pp.3-22
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• 1998

The effect of pH on degradation of paper by some fungi, which able to degrade cellulose, was investigated. Trichoderma koningii, Aspergillus nigerand Penicillium nigulosum were cultured at $28^{\circ}C$ for 16 days in the selective medium (PH3, PH4, PH5, PH6, PH7, PH8, PH9, PH10, PHC) containing paper as substrate. Each paper was pretreated with each pH buffer (pH 3∼pH 10, D.W.)prior to addition to the selective medium. Enzyme activities in the each culture medium were measured spectroph to metrically using C.M.C., Avicel, PNPG as the substrates for endoglucanase, exoglucanase and $\beta$-glucosidase, respectively. In all experimental fungi, the enzyme activities of PH3 and PH9 medium were usually much higher than those of other experimental groups. However in the PH6medium, enzyme activity was lower than other groups. To analyze the concentration and pattern of protein in the each culture medium, the medium was concentrated by lyophilization. The protein concentration of PH3 and PH9 medium were relatively high (T.koningii; 6.31mg, 6,19mg, A.niger; 1.62mg, 1.96mg, P.nigulosum;2.50mg, 2.73mg, respectively), but that of PH6 was relatively low. The protein pattern of each medium was analyzed by using SDS-PAGE and VDS Image Master Analysis Program. The concentrations of bands in the each lane were usually high at lane2 (PH3) and lane8 (PH9) and low at lane5 (PH6). Therefore, the incresed cellulolytic activity of fungus against acidified paper could be result of structural change and deterioration of paper caused by being acidified.

• The Korean Journal of Mycology
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• v.37 no.2
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• pp.181-188
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• 2009

A Gram-positive, rod-shaped bacteria named DY5 was isolated from a peat sample collected from Daeam mountain in Korea. The culture filtrate of the bacterial isolate DY5 showed a broad spectrum of antifungal activity on various crop pathogenic fungi such as Trichoderma koningii, Fusarium oxysporum, Colletotrichum gloeosporioides, Sclerotinia sclerotiorum, Rhizoctonia solani AG-1(IA) For the identification of the DY5, morphological, biochemical, API 50 CHB test, analysis of fatty acid and molecular phylogenetic approaches were performed. The DY5 was found to be a member of the genus Paenibacillus on the basis of morphological and biochemical analysis. The 16S rRNA of DY5 showed high similarity(98%) with Paenibacillus polymyxa. On the basis of these results, the DY5 was identified as Paenibacillus polymyxa. Antifungal substance of the DY5 would be mild alkaline proteine molecule. The DY5 seems to have a great potential to be a biocontrol agent against various crop pathogens.