• The Plant Pathology Journal
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• 제21권3호
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• pp.229-236
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• 2005

Molecular profIles of PCR-RFLP and sequence analysis of internal transcribed spacer (ITS) regions of rDNA were compared between morphologically distinguishable species of Trichoderma isolated from substrates of oyster mushroom in Korea, T. atroviride, T. citrinoviride, T. harzianum, T. longibrachiatum, T. virens, and two unidentified species, Trichoderma sp. 1 and 2. PCR­RFLP analysis divided the Trichoderma spp. into six RFLP groups, A, B, C, D, E, and F. The RFLP groups were generally agreed with described morphological species, except that the RFLP group A containing the two unidentified species. A neighbor-joining tree based on ITS sequences well supported RFLP groups observed by RFLP analysis of ITS regions of rDNA. Additionally, the two unidentified species, Trichoderma sp. 1 and 2, which could not be distinguished by PCR­RFLP analysis, were separated in sequence analysis of ITS regions of rDNA.

• Mycobiology
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• 제32권1호
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• pp.24-34
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• 2004

Molecular approaches, internal transcribed spacer(ITS) sequences of ribosomal DNA, and Universal Rice Primer Polymerase Chain Reaction(URP-PCR) were used to investigate the genetic diversity, taxonomic complexity, and relationships of Trichoderma species in mushroom farms. Forty-one isolates of 13 Trichoderma spp. were used in this study and clustered into eight groups. The DNA fingerprint patterns and ITS1 region sequence alignment data showed similar results, but not in some species, such as T. virens, T. atroviride, T. harzianum, and T. aureoviride. Results of this study have proven that the morphology-based taxonomic system has some limitations in terms of classification. The data obtained in this study would be a good index for classifying indistinguishable Trichoderma strains.

• Mycobiology
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• 제37권4호
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• pp.277-285
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• 2009

In this study, dual culture, poison agar, and direct methods were used to assess the ability of Trichoderma virens IMI-392430, T. pseudokoningii IMI-392431, T. harzianum IMI-392432, T. harzianum IMI-392433, and T. harzianum IMI-392434 to control Ceratocystis paradoxa, which causes the pineapple disease of sugarcane. The highest percentage inhibition of radial growth (PIRG) values were observed with T. harzianum IMI-392432 using two dual culture methods, 63.80% in Method I and 80.82% in Method II. The minimum colony overgrowth time was observed with T. harzianum IMI-392432 and the maximum was observed with T. pseudokoningii IMI-392431. Different concentrations of different day-old metabolites of Trichoderma isolates were tested against mycelial growth of C. paradoxa. The highest PIRG (84.685%) exhibited at 80% concentration of 30-day-old metabolites of T. harzianum IMI-392432 using the modified bilayer poison agar method. In the direct assay method the maximum mycelial growth weight (PIGW) was observed at the same concentration and the same day-old metabolites of T. harzianum IMI-392432. This study showed that Trichoderma isolates have a good antagonistic effect on C. paradoxa mycelial growth and T. harzianum IMI-392432 has the most potential to control the pineapple disease pathogen.

• Mycobiology
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• 제48권4호
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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).

• 한국식물병리학회지
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• 제11권4호
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• pp.292-297
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• 1995

We developed an in vitro assay method for evaluating plant growth promotion and providing an evidence that the growth promotion is rendered by growth enhancing factors. The amendment of culture filtrates of Trichoderma harzianum T95 and Gliocladium virens G872 and G872B in Murashige and Skoog (MS) agar medium enhanced the cotyledon growth of cucumber in terms of fresh weight and primary leaf area of cucumber cotyledon cuttings, of which the treatment of G. virens G872B was the most effective. The mycelial culture filtrate of G872B was more effective in the growth promotion than its conidial germling filtrate. The addition of 1% sucrose in MS mineral medium with 0.1% culture filtrates of the antagonists (T95 and G872B) was optimum for enhancing the effect of the filtrates on the growth of cotyledon cuttings in vitro. When cucumber seeds treated with G872B, Pseudomonas putida Pf3 or the G872B-Pf3 mixture were planted in a greenhouse, the rate of seed germination, biomass of shoot and root, and yield of cucumber fruits were increased, especially by G872B or the G872B-Pf3 mixture. Correspondingly, cucumber fruit yields in early to middle-cycles of harvest were significantly greater in the plots of G872B than the control and Pf3-treated plots, and the final yield was highest in the plots of the G872B-Pf3 mixture applications.

• The Plant Pathology Journal
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• 제26권1호
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• pp.32-36
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• 2010

Twelve fungal isolates out of 123 isolates obtained from cysts and soils of potato cyst nematode (PCN)-infested fields in Central Java, Indonesia had parasitic abilities of over 50% on PCN eggs or females (cysts) in vitro pathogenicity tests. Cultural and morphological characters and DNA sequences of ribosomal genes in ITS region revealed that they were four isolates of Gliocladium (Trichoderma) virens, three isolates of Fusarium oxysporum, one of F. lateritium, one of Penicillium tritinum and two of Taralomyces spp. A hundred percent infections occurred in eggs or cysts by three fungal isolates G. virens, F. oxysporum and P. oxalicum, suggesting that these fungi may have a good potential for the PCN biocontrol. Especially, G. virens isolates, which occurred most frequently in the PCN-infested potato fields and are known to be highly adaptable to varying habitats, may be developed as reliable agents for controlling PCN with both egg- and cyst-parasitic capabilities and with high ecological adaptabilities.

• Journal of Forest and Environmental Science
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• 제36권3호
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• pp.207-218
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• 2020

Ashwagandha is an important ancient medicinal crops, being affected with many diseases, among which leaf blight caused by Alternaria alternata has become the constraint resulting in huge yield losses. Continuous usage of chemical methods leads to environment, soil and water pollution. Whereas biological control of diseases is long lasting, inexpensive, eco-friendly and harmless to target organisms. In this context, it is aimed to evaluate five Trichoderma strains viz. Trichoderma virens IMI-392430, T. pseudokoningii IMI-392431, T. harzianum IMI-392432, T. harzianum IMI-392433 and T. harzianum IMI-392434 as bio-control efficacy against A. alternata and growth promoting effect in Ashwagandha. All the Trichoderma strains had varied antagonistic effects against the pathogen. In dual culture technique, the strain T. harzianum IMI-392433 showed maximum percentage inhibition of mycelial growth (54.89%) followed by T. harzianum IMI-392432 (53.83%), T. harzianum IMI-392434 (48.94%) and T. virens IMI-392430, (43.62%) against the pathogen, while the least inhibition percentage was observed with the T. pseudokoningii IMI-392431 (36.60%). The culture filtrate of the Trichoderma strain, T. harzianum IMI-392433 recorded highest inhibition on the mycelial growth (39.05%) and spore germination (80.77%) of pathogen and the lowest was recorded in T. pseudokoningii IMI-392431 (20.45 and 50%). Moreover, seeds treated with spore suspension of the strain T. harzianum IMI-392433 reduced the percentages of disease severity index significantly. The strain T. harzianum IMI-392433 also significantly increased seed germination %, seedling vigor and growth of Ashwagandha. The correlation matrix showed that root yield per plant of Ashwagandha had significant and positive correlation with plant height (r=0.726^⁎⁎), number of leaf (r=0.514^⁎⁎), number of primary branch (r=0.820^⁎⁎), number of secondary branch (r=0.829^⁎⁎), fresh plant weight (r=0.887^⁎⁎), plant dry weight (r=0.613^⁎⁎), root length (r=0.824^⁎⁎), root diameter (r=0.786^⁎⁎), root dry weight (r=0.739^⁎⁎) and fresh root weight (r=0.731^⁎⁎). The significant and negative correlation (r=-0.336^⁎⁎) was observed with the root yield and percentages of disease severity index. The study recognized that the T. harzianum IMI-392433 strain performed well in inhibiting the mycelial growth and reduced the percentages of disease severity index of pathogen as well as increased the plant growth in Ashwagandha.

• The Plant Pathology Journal
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• 제18권5호
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• pp.279-283
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• 2002

Biological control of Fusarium udum causing wilt disease of pigeonpea was studied in vitro, as well as, in vivo. Aspergilluspavus, Anergillus niger, Bacilius licheniformis (strain-2042), Gliocladium virens, Peniciliium citrimum, and Trichoderma harzianum, which were found to be the most potent ones in inhibiting the radial colony growth of the test pathogen, were used as biological control by amending their inocula at diffeyent concentrations in pots and in pathogen-infested soil in the fields. Maximum reduction of the wilt disease was observed with G. vireos both in pots and in the fields. The population of E. udum was found to be markedly reduced when the antagonists were applied in the soil. The study establishes that G. virens can be exploited for the biological control of wilt disease at field level.

• Mycobiology
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• 제38권2호
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• pp.113-117
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• 2010

Trichoderma strains were evaluated under field conditions to assay their efficacy in suppressing Alternaria fruit rot disease and promoting chili plant growth. The experiment was conducted at the Botanical Garden, Rajshahi University, Bangladesh from July 2006 to March 2007. Application of Trichoderma harzianum IMI 392432 significantly (p = 0.05) suppressed the disease compared to Alternaria tenuis (T2) treatment and improved both growth and yield. The treatment T4 (T. harzianum IMI-392432 + A. tenuis) was most effective in reducing disease percentage (72.27%) compared to A. tenuis (T1) treatment. The highest seed germination rate (85.56%) and the highest growth and yield (12.5 g/plant) was also recorded in the same treatment (T4), followed by T5 (T. harzianum IMI-392433 + A. tenuis), T6 (T. harzianum IMI-392434 + A. tenuis), T2 (T. virens IMI-392430 + A. tenuis), and T3 (T. pseudokoningii IMI-392431 + A. tenuis) treatment, while single treatment with A. tenuis significantly decreased these values.

• 한국버섯학회지
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• 제12권3호
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• pp.201-208
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• 2014

버섯배지의 발효를 효율적으로 행하면서 버섯의 재배 시 빈번하게 발생하는 푸른곰팡이 병의 원인 균주인 Trichoderma sp. 곰팡이 성장을 억제하는 세균의 분리를 행하였다. 균원시료로부터 1차 분리한 약 200여 균주 중에서 성장속도가 빠르고, SM, AM 및 CM의 평판배지 상에서 clear zone이 뚜렷한 6균주를 2차 분리하였다. 분리한 6균주 중 cellulase, amylase 및 protease의 효소활성이 높고 T. virens와 T. harzianum에 대하여 강한 항균활성을 나타낸 C-1균주를 최종 분리균주로 선정하였다. 분리한 C-1균주는 Bergey's Manual of Systematic Bacteriology에 의한 동정과 16S rDNA 염기서열 분석을 행한 결과 Paenibacillus polymyxa 밝혀져 P. polymyxa CK-1으로 명명하였다. P. polymyxa CK-1균주의 생육조건을 검토한 결과 최적배양온도는 $45^{\circ}C$, 생육을 위한 배지의 최적 pH는 6.0~7.0 범위로 나타났다. T. virens와 T. harzianum 곰팡이의 생육억제를 위한 P. polymyxa CK-1의 배양시간은 22~36시간이 적당하였다. 그리고 P. polymyxa CK-1균주의 24시간째 배양용액을 처리한 petri dish에 두 곰팡이를 각각 접종한 후 10일 동안 방치하여도 곰팡이의 생육은 관찰되지 않았다. P. polymyxa CK-1 균주가 생산한 길항물질의 열안정성을 검토한 결과 $60^{\circ}C$ 및 $100^{\circ}C$로 20분 동안 처리한 시험구에서는 두 곰팡이의 균사성장이 전혀 관찰되지 않았지만 $121^{\circ}C$에서 20분 동안 처리한 시험구에서는 약간의 균사성장이 관찰되었다. P. polymyxa CK-1배양액이 버섯균사 생육에 미치는 영향을 검토한 결과 팽이버섯, 표고버섯 등의 다양한 버섯균사 생육에 전혀 영향을 미치지 않는 것으로 나타났다.