• Title/Summary/Keyword: Trichoderma harzianum.

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Partial Purification of Lectin from Mycoparasitic Species of Trichoderma

  • Singh, Tanuja;Saikia, Ratul;Arora, Dilip K.
    • The Plant Pathology Journal
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    • v.21 no.4
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    • pp.301-309
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    • 2005
  • Trichoderma species/isolates exhibited varied degree of agglutination on sclerotial (Sc) and hyphal (Hy) surface of Macrophomina phaseolina. The agglutination efficiencies on Sc and Hy ranged from $11\;to\;57\%$. Isolates of T. harzianum (Th) and T. viride (Tv) showed greater agglutination on Sc ($23-57\%$) and Hy ($16-47\%$). Different enzymes (trypsin, pepsin, proteinase k, a-chymotrypsin, lyticase and glucosidase) and inhibitors (tunicamycin, cycloheximide, brefeldin A, sodium azide, dithiothreitol and SDS) reduced the agglutination potential of conidia of Th-23/98 and Tv-25/98; however, the extent of response varied greatly in different treatments. Different fractions of Th-23/98 and Tv-25/98 exhibited haemagglutinating reaction with human blood group A, B, AB and O. Haemagglutinating activity was inhibited by different sugars and glycoproteins tested. Crude haemagglutinating protein from outer cell wall protein fraction of Th-23/98 and Tv-25/98 were eluted on Sephadex G-100 column. Initially Th-23/98 and Tv-25/98 exhibited two peaks showing no agglutination activity; however, lectin activity was detected in the third peak. Similar to crude lectin, the purified lectin also exhibited haemagglutinating activity with different erythrocyte source. SDS-PAGE analysis of partially purified lectin revealed single band with an estimated molecular mass of 55 and 52 kDa in Th-23/98 and Tv-25/98, respectively. Trypsin, chymotrypsin and b-1,3-glucanase totally inhibited lectin activity. Similarly, various pH also affected the haemagglutinating activity of Th-23/98 and Tv-25/98. From the present observations, it can be concluded that the recognition/attachment of mycoparasite (T. harzianum and T. viride) to the host surface (M. phaseolina) may be most likely due to lectin-carbohydrate interaction.

Trichoderma asperellum Chi42 Genes Encode Chitinase

  • Loc, Nguyen Hoang;Quang, Hoang Tan;Hung, Nguyen Bao;Huy, Nguyen Duc;Phuong, Truong Thi Bich;Ha, Tran Thi Thu
    • Mycobiology
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    • v.39 no.3
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    • pp.182-186
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    • 2011
  • Four Trichoderma strains (CH2, SH16, PQ34, and TN42) were isolated from soil samples collected from Quang Tri and Thua Thien Hue provinces in Vietnam. The strains exhibited high chitinolytic secretion. Strain PQ34 formed the largest zone of chitinase-mediated clearance (> 4 cm in diameter) in agar containing 1% (w/v) colloidal chitin. Analysis of the internal transcribed spacer regions of these strains indicated that they were Trichoderma asperellum. The molecular weights of the chitinases were approximately 42 kDa. Chitinase genes (chi42) of T. asperellum strains TN42, CH2, SH16, and PQ34 were 98~99% homologous to the ech42 gene of T. harzianum CB-Pin-01 (accession No. DQ166036). The deduced amino acid sequences of both T. asperellum strains SH16 and TN42 shared 100% similarity.

Cytotoxicity of Trichoderma spp. Cultural Filtrate Against Human Cervical and Breast Cancer Cell Lines

  • El-Rahman, Atef Abd El-Mohsen Abd;El-Shafei, Sally Mohamed Abd El-Aziz;Ivanova, Elena Vladimirovna;Fattakhova, Alfia Nurlimanovna;Pankova, Anna Victorovna;El-Shafei, Mohamed Abd El-Aziz;El-Morsi, El-Morsi Abu El-Fotouh;Alimova, Farida Kashifovna
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7229-7234
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    • 2014
  • Trichoderma spp. are known as a rich source of secondary metabolites with biological activity belonging to a variety of classes of chemical compounds. These fungi also are well known for their ability to produce a wide range of antibiotic substances and to parasitize other fungi. In search for new substances, which might act as anticancer agents, the overall objective of this study was to investigate the cytotoxic effects of Trichoderma harzianum and Trichoderma asperellum cultural filtrates against human cervical and breast cancer cell lines (HeLa and MCF-7 cells respectively). To achieve this objective, cells were exposed to 20, 40, 60, 80 and 100 mg/ml of both T. harzianum cultural filtrate (ThCF) and T. asperellum cultural filtrate (TaCF) for 24h, then the cell viability and the cytotoxic responses were assessed by using trypan blue and 3-(4,5-dimethylthiazol-2yl)-2,5-biphenyl tetrazolium bromide (MTT) assays. Morphological changes in cells were investigated by phase contrast inverted microscopy. The results showed that ThCF and TaCF significantly reduce the cell viability, have cytotoxic effects and alter the cellular morphology of HeLa and MCF-7 cells in a concentration dependent manner. A concentration of 80 and 100mg/ml of ThCF resulted in a sharp decline in the cell viability percent of HeLa and MCF-7 respectively (25.2%, 26.5%) which was recorded by trypan blue assay. The half-maximal inhibitory concentrations ($IC_{50}$) of ThCF and TaCF in HeLa and MCF-7 were recorded as 16.6, 12.0, 19.6 and 0.70mg/ml respectively by MTT assay. These results revealed that ThCF and TaCF have a substantial ability to reduce the viability and proliferation of human cervical and breast cancer cells.

Characterization of a New Cultivar, 'Cham' in Agrocybe aegerita (버들송이 신품종 '참'의 특성)

  • Jeon, Dae-Hoon;Choi, Jong-In;Ha, Tai-Moon;Ju, Young-Cheol
    • Korean Journal of Breeding Science
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    • v.42 no.3
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    • pp.288-293
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    • 2010
  • 'Cham', a new cultivar of Agrocybe aegerita, was bred by crossing between monokaryotic strains isolated from GMAG45021 and 'Beodeulsongi1ho' in Mushroom Research Station, Gyonggi Province A.R.E.S. in 2007. The optimum temperature for the mycelial growth of 'Cham' was 26 to $28^{\circ}C$ on PDA medium, whereas that of 'Beodeulsongi1ho'(control) was 24 to $26^{\circ}C$. The optimum temperature for the primordial formation and fruiting body development of 'Cham', as well as 'Beodeulsongi1ho', was 18 to $20^{\circ}C$. In the bottle cultivation, the spawn run period of 'Cham' at 22 to $23^{\circ}C$ was around 38 days and the period from scratching of inoculum to harvest was 12 days. These characteristics of 'Cham' were not different from those of 'Beodeulsongi1ho'. However, 'Cham' had more dark brown-colored, and thicker and stronger pileus, and longer and thicker stipe compared to that of 'Beodeulsongilho'. Freshness of 'Cham' was maintained for 10 days at the storage temperature of $4^{\circ}C$, while that of 'Beodeulsongi1ho' was maintained for 8 days. Moreover, the yield of fruiting bodies of 'Cham' was $141g/850m{\ell}$ bottle, which was similar to that of 'Beodeulsongi1ho'. Both 'Cham' and 'Beodeulsongilho' have lower resistivity against Trichoderma virens and Trichoderma harzianum.

Isolation, Identification and Antagonisms of Rhizospheric Antagonists to Cucumber Wilt Pathogen, Fusarium oxysporum f. sp. cucumerinum Owen (오이 덩굴쪼김병균에 대한 오이 근권길항미생물의 분리, 동정 및 길항작용)

  • Jee Hyeong Jin;Kim Hee Kyu
    • Korean Journal Plant Pathology
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    • v.3 no.3
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    • pp.187-197
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    • 1987
  • Bacteria and fungi antagonistic to Fusarium oxysporum f. sp. cucumerinum Owen were effectively isolated with each of modified Triple Layer Agar (TLA) technique from rhizosphere soil where cucumber had been grown healthily in plastic film house. Three predominant bacterial isolates selected were identified as Pseudomonas fluorescens, and P. putida, Serratia sp. and three fungal isolates were Gliocladium sp. Trichoderma harzianum, and T. viride. Antagonistic bacteria inhibited $26-45\%$ of germination and $41-56\%$ of germ tube elongation of microconidia of F. oxysporum f. sp. cucumerinum on Water Agar (WA). P. fluorescens was the strongest inhibitor. Several my co parasitisms were observed on dual culture of WA between antagonistic fungi and F. oxysporum f. sp. cucumerinum such as coiling, penetration, overgrowing, and lysis. Mycelial lysis of the pathogen was the most severe at pH 4.6, followed by 3.6, 5.6 and 6.6 of the medium in decreasing order. At pH 6.6, mycelia of the pathogen were not conspicuously damaged, however, the antagonistic fungi formed abundant chlamydospores especially Gliocladium sp. T. harzianum revealed the most excellent antagonism in vitro.

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Isolation, Identification, and Evaluation of Biocontrol Potentials of Rhizosphere Antagonists to Rhizoctonia solani (원예작물(園藝作物) 모잘록병(Rhizoctonia solani $K\"{u}hn$)의 발생(發生)에 관여하는 근권길항균(根圈拮抗菌)의 분리(分離), 동정(同定) 및 생물적(生物的) 방제(防除) 검토(檢討))

  • Kim, Hee-Kyu;Roh, Myung-Ju
    • Korean journal of applied entomology
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    • v.26 no.2 s.71
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    • pp.89-97
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    • 1987
  • Antagonistic microorganisms from rhizosphere soil were isolated, identified, and applied successfully as the biocontrol agents of damping-off caused by Rhizoctonia spp. Rhizosphere antagonists isolated from rhizosphere soil were identified as Trichoderma viride, T. harzianum, T. hamatum, T. polysporum, Gliocladium sp., Pseudomonas fluorescence, P. stutzeri, P. cepacia, Enterobacter sp., Serratia sp. and Erwinia herbicola. Of these, the most promising ones in vitro were T. virdie, T. harzianum, Gliocladium sp., Serratia sp., P. stutzeri, and P. cepacia. These above six antagonists were efficient in reducing disease incidence to $40{\sim}70%$ when the reselected rhizosphere antagonists preparations were applied to the soil at $10^6$ propagules per gram. Among six antagonists, T. viride was the most promising biocontrol agents against R. solani isolates in soil. The suppressive effect was more evident in steam-sterilized soil than in non-sterilized field soil.

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Analysis of Fungal Concentration and Species Present as Bio-aerosols in Oak Mushroom Cultivation Houses (국내 표고버섯 재배사에 바이오에어로졸로서 분포하는 진균의 농도와 종 분석)

  • Kim, Seong Hwan;Kim, Ji Eun;Kim, Jun Young
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.393-403
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    • 2018
  • Bio-aerosols transported by the air have been considered as the major source of dispersal agents that contaminate agricultural products. Unseen fungal spores are known sources of bio-aerosols that harm mushroom and human health during mushroom cultivation. This study was conducted to obtain basic data on the concentration and species distribution of fungi present in the indoor air of oak mushroom cultivation houses in Korea. In 2015 and 2016, we sampled and analyzed indoor airborne fungal spores 21 times from 13 oak mushroom cultivation farms located in six different provinces. The concentration of airborne fungi ranged from $1.30{\times}10^2$ to $1.59{\times}10^4cfu/m^3$. Surprisingly, in 20 sampling cases, the fungal concentration exceeded $500cfu/m^3$, which is recommended as the indoor air quality standard by the Ministry of Environment, Korea. A total of 450 fungi were isolated and identified to belong to 33 genera and 46 species. Among the identified fungi, human pathogens (4 genera and 4 species) and plant pathogens (10 genera and 13 species) were present. In addition, Trichoderma harzianum, Trichoderma atroviride, and Trichoderma longibrachiatum, which are detrimental species that affect mushroom health, were found 17 out of 21 sampling times. Our results provide evidence that indoor air quality should be improved for better management of mushroom cultivation houses.

Rice Straw-Decomposing Fungi and Their Cellulolytic and Xylanolytic Enzymes

  • Lee, Sang-Joon;Jang, Yeong-Seon;Lee, Young-Min;Lee, Jae-Jung;Lee, Han-Byul;Kim, Gyu-Hyeok;Kim, Jae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1322-1329
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    • 2011
  • Filamentous fungi colonizing rice straw were collected from 11 different sites in Korea and were identified based on characterization of their morphology and molecular properties. The fungi were divided into 25 species belonging to 16 genera, including 14 ascomycetes, one zygomycete, and one basidiomycete. Fungal cellulolytic and xylanolytic enzymes were assessed through a two-step process, wherein highly active cellulase- and/or hemicellulase-producing fungi were selected in a first screening step followed by a second step to isolate the best enzyme-producer. Twenty-five fungal species were first screened for the production of total cellulase (TC), endo-${\beta}$-1,4 glucanase (EG), and endo-${\beta}$-1,4 xylanase (XYL) using solid-state fermentation with rice straw as substrate. From this screening, six species, namely, Aspergillus niger KUC5183, A. ochraceus KUC5204, A. versicolor KUC5201, Mucor circinelloides KUC6014, Trichoderma harzianum 1 KUC5182, and an unknown basidiomycete species, KUC8721, were selected. These six species were then incubated in liquid Mandels' media containing cellulose, glucose, rice straw, or xylan as the sole carbon source and the activities of six different enzymes were measured. Enzyme production was highly influenced by media conditions and in some cases significantly increased. Through this screening process, Trichoderma harzianum 1 KUC5182 was selected as the best enzyme producer. Rice straw and xylan were good carbon sources for the screening of cellulolytic and xylanolytic enzymes.

Integrated Control of Large Patch Disease caused by Rhizoctonia solani AG2-2 by Using Fertilizers, Fungicides and Antagonistic Microbes on Turfgrasses (잔디에 발생하는 라지패취병의 종합적 방제)

  • 심규열;김희규;배동원;이준택;이현주
    • Asian Journal of Turfgrass Science
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    • v.11 no.3
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    • pp.173-183
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    • 1997
  • This study was conducted to develop an integrated disease management system against large patch disease. Attempts were made to estimate the effect of calcium hydroxide, silicate fertilizer and urea on the mycelial growth of Rhizoctonia sotani AG2-2 and the development of large patch in vitro and in vivo and to establish the most promising combination of fertilizer, fungicide and antagonistic microbes. 1.The mycelial growth of Rhizoctonia sotani AG2-2 were completely inhibited at 2,000, 1,000 and 3,000ppm concentration by calcium hydroxide, silicate fertilizer and urea, respectively. Inhibition effect of silicate fertilizer was the highest, but that of urea was the lowest compared with other treatments. 2.Treatment of calcium hydroxide at rate of l00g /$m^2$ was the most effective, and control effect appeared from 30 days after treatment in spring, which was better in autumn than in spring. Silicate fertilizer treated at rate of l00g /$m^2$, 200g /m$^2$, in spring and 50g /$m^2$ in autumn were very effective. Urea at rate of 30g /$m^2$ was more effective than 60g /m$^2$ and 120g /m$^2$. 3.The efficacies of mepronil and toclofos-methyl, applied twice in spring and once in autumn, were 83.8% in spring, which persisted to 70% in autumn compared with untreated plot. 4.The efficacies of Trichoderma harzianum were the highest by 55% in spring, but those of Bacillus amyloliquefaciens BL-3 and Peudomonas putida were the highest by 80% in autumn among other antagonists tested, when two organisms were applied twice in spring and once in autumn.

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Diversity analysis of culture-dependent fungal species isolated from the sawdust media of Lentinula edodes (표고 톱밥배지에서 분리한 배양 진균의 다양성 분석)

  • Kim, Min-Keun;Sim, Soon-Ae;Choi, Si-Lim;Hong, Kwang-Pyo
    • Journal of Mushroom
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    • v.17 no.4
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    • pp.179-184
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    • 2019
  • This study was conducted to investigate the diversity of culture-dependent fungal species in the sawdust media of Lentinula edodes. A total of 405 fungi were isolated from the specimens and identified to belong to 24 genera and 42 species. Among the identified 42 species of fungi, 26.2% belonged to Penicillium sp., 9.5% belonged to Trichoderma sp., and 64.3% belonged to others. Especially, Trichoderma harzianum, which is a causal agent of fungal disease in mushroom, was found on all the farms, and showed the highest frequency among the identified fungi. Community analysis showed that the fungal diversity patterns of the samples were similar to each farm and many kinds of fungi existed in the sawdust media at high levels. These results showed that the management of internal environments would be required for the stable cultivation of Lentinula edodes.