• Journal of Practical Agriculture & Fisheries Research
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• v.24 no.3
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• pp.5-14
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• 2022

Trichoderma is known as pathogen caused serious green mold disease on commercial production. T. pleuroti and T. pleuroticola were common species in various mushroom media. Many strains of T. pleuroti, known as aggressive species causing major economic losses in Korea, showed benomyl resistance. Accurate identification and detection of Trichoderma species associated with oyster mushrooms is very important for disease control. We developed species-specific primers for T. pleuroticola, T. pleuroti, T. harzianum, and T. atroviride based on species-specific fragments isolated from amplified fragment length polymorphism analysis. PCR products corresponding to the predicted fragment of 500bp, 230bp, 180bp, and 410bp were amplified from T. pleuroticola, T. pleuroti, T. harzianum, and T. atroviride, respectively. Multiplex PCR assay using species-specific primers quickly and accurately identified and detected T. pleuroti from mushroom media in which various species co-exist. Our results can be useful for the effective control of mushroom disease.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.127.1-127
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• 2003

We describe two new Trichoderma species associated with oyster mushroom in Korea. Trichoderma green mould has been one of the most serious diseases of oyster mushroom in Korea. Of these the predominant species are two unrecorded species. We designed as Trichoderma sp. Korean type 1 (Th K1) and Trichoderma sp. Korean type 2 (Th K2), respectively. Th K1 and Th K2 can be distinguished from previously reported Trichoderma species as well as each other in morphological characteristics including growth rate at 35$^{\circ}C$, colony morphology, conidia shape and branch pattern of phialides. Sequence of the ITS region of rDNA, the protein coding translation elongation factor gene(EF-1${\alpha}$), and RNA polymeraseII (RPB2) not only clearly separated Trichoderma sp. Korean types from their closely related T. harzianum biotype but also distinguished them from each other. Analyses of the EF-1${\alpha}$ and RPB2 sequences were found to be more useful for establishing systematic relationships among Trichoderma isolates than those of the ITS sequence. Based on the results of morphological and molecular characteristics. We propose the two Trichoderma sp. Korean types as the new species

• The Korean Journal of Pesticide Science
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• v.14 no.4
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• pp.401-406
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• 2010

This study was conducted to investigate the control effects of the environmental-friendly materials against major diseases of pear and to elucidate factors related to spore disperse of pear scab and pear rust. Spore disperse was deeply related ambient temperature in the early stage of disease occurrence and the maximum spore disperse was affected greatly by the air humidity, Percent of infected leaves of pear scab in plot treated with Trichoderma harzianum-YC459 was 29.3% and 43.3% in plot treated with Sulfur-80%. Two formulations above-mentioned were less effective to control pear scab than the chemical pesticide, hexaconazole. Control effect in plots single- and mixed-treated with Trichoderma harzianum-YC459, Sulfur-80% and hexaconazole against pear scab were compared with that in plot treated with chemicals by conventional application schedule in pear orchard. The incidence of pear scab was lower in plot treated with the environmental-friendly materials than that of untreated plot. However there was no significant difference among the treatments at the 5% level. Application of Sulfur-80% reduced significantly the incidence of pear scab (7.8%) compared to that (10.3%) in plot treated by conventional application schedule in organically cultivated pear orchard. Alternative application of Trichoderma harzianum-YC459 and Sulfur-80% (4.3%) suppressed the incidence of pear rust than that (7.1%) in plot treated by conventional application schedule in organically cultivated pear orchard.

• Journal of Microbiology and Biotechnology
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• v.21 no.8
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• pp.808-817
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• 2011

Because of its elevated cellulolytic activity, the filamentous fungus Trichoderma harzianum has a considerable potential in biomass hydrolysis applications. Trichoderma harzianum cellobiohydrolase I (ThCBHI), an exoglucanase, is an important enzyme in the process of cellulose degradation. Here, we report an easy single-step ion-exchange chromatographic method for purification of ThCBHI and its initial biophysical and biochemical characterization. The ThCBHI produced by induction with microcrystalline cellulose under submerged fermentation was purified on DEAE-Sephadex A-50 media and its identity was confirmed by mass spectrometry. The ThCBHI biochemical characterization showed that the protein has a molecular mass of 66 kDa and pI of 5.23. As confirmed by smallangle X-ray scattering (SAXS), both full-length ThCBHI and its catalytic core domain (CCD) obtained by digestion with papain are monomeric in solution. Secondary structure analysis of ThCBHI by circular dichroism revealed ${\alpha}$- helices and ${\beta}$-strands contents in the 28% and 38% range, respectively. The intrinsic fluorescence emission maximum of 337 nm was accounted for as different degrees of exposure of ThCBHI tryptophan residues to water. Moreover, ThCBHI displayed maximum activity at pH 5.0 and temperature of $50^{\circ}C$ with specific activities against Avicel and p-nitrophenyl-${\beta}$-D-cellobioside of 1.25 U/mg and 1.53 U/mg, respectively.

• The Korean Journal of Mycology
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• v.31 no.1
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• pp.44-49
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• 2003

Several microbial biocontrol products (Greenbiotech Co., Paju, Korea), Green-all T (Trichoderma harzianum), Green-all S(Bacillus sp.) and Green-all G (Streptomyces sp.) were supplemented with organic amendments such as sawdusts and rice hulls to study on efficacy of biological control of seeding damping-off of cucumber caused by Pythium ultimum. Sawdusts amended into potato dextrose agar alone could inhibit in vitro mycelial growth of P. ultimum. All there microbial products of Green-all T, Green-all G and Green-all S significantly reduced seeding damping-off (LSD, P=0.05). However, several amendments such as sawdusts and rice hulls into Green-all T and Green-all S products did not increase efficacy of biological control compared to non-amended treatment. In contrast, supplements of aminodoctor containing several amino acids (Greenbiotech Co., Korea) into Green-all G product significantly increased efficacy of biological control of seeding damping-off, resulting in from 42% to 2% disease incidence in relation to seedling emergence (LSD, P=0.05). Also, amendment of sawdusts into Tricoderma product significantly increased efficacy of biological control as disease index of 5.0 compared to non-amended control of 56.0 in Green-all T product alone. This indicates that organic amendments could increase efficacy of biological control of cucumber seedling damping-off.

• The Plant Pathology Journal
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• v.28 no.2
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• pp.137-148
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• 2012

During investigating of shiitake mushroom competitors, 289 isolates of Trichoderma spp. were collected from shiitake mushroom farms in different districts and the Forest Mushroom Research Center of Korea, among which 29 representative strains were selected. Based on the DNA sequences of the rpb2 and tef1 genes and the ITS rDNA, and their morphological characteristics, they were identified as T. atroviride, T. citrinoviride, T. harzianum, T. longibrachiatum, and two undescribed species, Trichoderma spp. 1 and 2, which are considered to be the candidate of new species. Competition tests between Lentinula edodes (Sanjo302) and the Trichoderma species indicated that the six species of Trichoderma were significantly different from each other in terms of their ability to invade the mycelial blocks of shiitake. In both of dual cultures on potato dextrose agar and sawdust media, Trichoderma spp. 1 and 2 strongly invaded the mycelial blocks of shiitake. Our results suggest that the two Trichoderma species may cause potentially serious economic losses in shiitake cultivation of Korea.

• Mycobiology
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• v.33 no.3
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• pp.147-149
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• 2005

The investigation was undertaken to enhance the decomposition process by pre-treatment of rice stubble, having higher concentration of lignin. Air-dried rice stubble was treated with 1.8 liter of 1% NaOH and autoclaved. Six cellulolytic fungi, Trichoderma harzianum, Penicillium citrinum, Curvularia lunata, Aspergillus flavus and Alternaria alternata were grown in basal synthetic medium along with delignified rice-residue as carbon source for production of soluble crude protein. Though the loss of cellulose has been observed by all of them but having a considerable status in the presence of T. harzianum and T. harzianum yielded highest percentage of crude protein (27.99%) with biomass of 375 mg, whereas the lowest protein value (17.91%) was recorded in case of A. niger with biomass of 422 mg. Among the imperfect fungi, T. harzianum was the most potent. Effects of incubation period and nitrogen sources on soluble crude protein production by T. harzianum were also undertaken in this study. Fifth day of incubation period and potassium nitrate as nitrogen source among other nitrogen sources was found most appropriate for soluble crude protein production by the mentioned organism.

• Journal of the Korean Wood Science and Technology
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• v.37 no.6
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• pp.585-599
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• 2009

This study was carried out to investigate the antifungal activity of the water soluble essential oil against Trichoderma spp. Water soluble essential oils from Pinus densiflora and Chamaecyparis obtusa were obtained from GAP (Gas assisted process) extraction apparatus. When the water soluble essential oil was treated, the growth of the conidial germination was maximum under the cultivation condition at $25^{\circ}C$ and in the culture medium adjusted to pH 5.0. The yield of water soluble essential oil was 3.9% and 3.7% in P. densiflora and C. obtusa, respectively. 24 and 15 kinds of compounds were identified in water soluble essential oils of P. densiflora and C. obtusa, respectively. The major components in the essential oil of P. densiflora were $\alpha$-Terpineol acetate and Terpinen-4-ol and those of C. obtusa were Terpinen-4-ol and $\alpha$-Terpineol. Antifungal activity was the best in the 5000 ppm of P. densiflora against Trichoderma harzianum and in 5000 ppm of C. obtusa against Trichoderma atroviride.

• Journal of agriculture & life science
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• v.43 no.3
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• pp.15-26
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• 2009

This study was undertaken to determine inhibitory compounds from extracts of the softwood (larix leptolepis, Pseudotsuga menziesii) sawdust against Trichoderma spp. The sawdust of L. leptolepis and P. menziesii were hot water extracted, which were with fraction extracted organic solvents. The organic solvent extractions were carried out by n-hexane, methylene chloride, ethyl acetate. The antifungal activity of hot water extracts of L. leptolepis sawdust was determined to be 20.6% inhibition at a concentration of 1,000 ppm against Trichoderma spp. The antifungal activity of P. menziesii sawdust was outstanding about 60.3% against Trichoderma spp. The yields of the fractions of n-hexane soluble, methylene chloride soluble and ethyl acetate soluble from the hot water extract of L. leptolepis sawdust were 4.0%, 6.0% and 8.0%, repectively. However, the yields of the fractions of three solvents of P. menziesii sawdust were 8.0%, 13.0 and 14.0% correspondingly. The antifungal activity of n-hexane soluble fraction from hot water extracts of L. leptolepis sawdust was highest to about 68.5% to 79.9% against Trichoderma spp. compared to others. The antifungal activities of n-hexane soluble fraction from hot water extracts of P. menziesii sawdust showed 68.5%, 71.4%. 71.9%, 75.7% and 82.3% against T. aggressivum, T. atroviride, T. harzianum, T. koningii and T.viride, respectively. The n-hexane soluble fraction revealed much higher antifungal activity than the other fractions did. This study demonstrated that the n-hexane fraction of the hot water extracts of L. leptolepis and P. menziesii exhibited the greatest antifungal activity against Trichoderma spp.

• Journal of Mushroom
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• v.12 no.3
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• pp.201-208
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• 2014

The separation of the bacteria inhibiting Trichoderma sp. mold, the strain causing blue mold disease that occurs frequently when cultivating mushroom while carrying out the efficient fermentation of mushroom medium, from the growth was done. In about 200 strains isolated primarily from fungus garden samples, 6 strains were secondly isolated, which had fast growth rates and a clear zone on the plate medium of SM, AM, and CM. Among the 6 strains isolated, the C-1 strain showed high enzymatic activity of cellulase, amylase, and protease, and strong antibacterial activity for the T. virens and T. harzianum, selected finally. The selected C-1 strain was identified as Paenibacillus polymyxaby the result of the identification by Bergey's Manual of Systematic Bacteriology and the analysis of the nucleotide sequence of 16S rRNA, and named as P. polymyxa CK-1. In reviewing the growth conditions of the P. polymyxa CK-1 strain, the optimum cultivation temperature was $45^{\circ}C$, and the optimum pH for growth was in the range of 6.0~7.0. Appropriate incubation time of P. polymyxa CK-1 for the growth inhibition of the fungus T. virens and T. harzianum was 22 to 36 hours. And the fungal growth was not observed, even when leaving two molds inoculated on each petri dishes, which were treated with 24 hour culture solution of P. polymyxa CK-1 strain for 10 days. As a result of studying the thermal stability of the antagonists produced by the P. polymyxa CK-1 strain, no mycelial growth of the two fungi was observed in the test group treated for 20 minutes at $60^{\circ}C$ and $100^{\circ}C$, but mycelial growth was slightly observed in the test group treated for 20 minutes at $121^{\circ}C$. As aresult of reviewing the impact of the P. polymyxa CK-1 culture medium on mushroom mycelial growth, it showed no effect on a variety of mushroom mycelial growth including enoki mushroom and shiitake mushroom.