• 미생물학회지
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• 제31권2호
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• pp.157-165
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• 1993

A 1, 4-.betha.-D-xylanase, designated as xylanase II, was purified from the culture filtrate of Trichoderma koningii ATCC 251131 by column chromatography on Sephadex G-75, SP-Sephadex C-50, DEAE-Sephadex A-50 and Sephadex G-50 with an overall yield of 6.97%. It has a molecular weight of 21.000 and an isoelectric point of 9.4. The enzyme activity is optimal at pH 5.0 and at a temperature of 50.deg.C. Xylanase II is stable up to 50.deg.C, while 40 and 90% of its activity are lost after the incubation for 30 and 60 min at 60.deg.C. The enzyme degrades xylan with relatively high activity, as well as carboxymethylcellulose and Avicel. Its $K_{m}$ values for oat-spelt xylan, larchwood xylan and Avicel are 7.48, 1.98 and 13.33 mg/ml, respectively. The hydrolysis products of oat-spelt xylan by xylanase II are xylose, xylobiose, xylotriose and arabinoxylotriose, while the reaction products of larchwood xylan are xylose, xylobiose, xylotriose and small amount of higher oligomers. The action paterns of the enzyme demonstrate that xylanase II is endo-enzyme.

• 미생물학회지
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• 제27권1호
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• pp.35-42
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• 1989

The mode of transglycosylation reaction observed during the action of low-molecular-weigh $\beta$-D-glucosidase ($\beta$-D-glucoside glucohydrolase, EC3.2.1.21) purified from Trichoderma koningii ATCC 26113 was investigated using $^{1}H$-NMR spectroscopy. The enzyme was purified by the series of procedures including ammonium sulfate precipitation, and fractionations by column chromatographies on Bio-Gel P-150, DEAE-Sephadex A-50, and SP-Sephadex C-50. The final purification was performed by the band eluation after preparative polyacrylamide gel electrophoresis. The enzyme showed its molecular size of 78,000 through the analysis of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and its isoelectric point of 5.80 through the analysis of analytical isoelectric focusing. The H-1 proton resonances were analyzed. After the reaction of the enzyme with cellobiose, the reaction products were separated by high performance liquid chromatography using refractive index detector. H-1 resonances of the products were consisted with those of gentiobiose [$\beta$-D-glucopyranosyl--(1,6)-D-glucopyranose], and cellotriose [$\beta$-D glucopyranosyl-(1,4)-$\beta$-D-glucopyranosyl]-(1,4)-D-glucopyranose] with minor resonances of sophorose [$\beta$-D-glucopyranosyl-(1,2)-D-glucopyranose], respectively.

• 식물병과 농업
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• 제1권1호
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• pp.19-29
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• 1995

A microbial product composed of three antagonistic fungal isolates (Aspergillus sp., Penicillium sp. and Trichoderma sp.) and three bacterial isolates (Arthrobacter sp., Bacillus sp., and Pseudomonas sp.) was tested for the control of Pythium blight caused by Pythium sp., brown patch by Rhizoctonia solani (anastomosis group(AG) 1-1) and large patch by R. solani (AG 2-2) of turfgrasses. Cultures of the antagonistic fungi and bacteria varied in the effectiveness in reducing disease severity of Pytium blight and brown patch on bentgrass. The antagonistic fungal and bacterial isolates were mixed and cultured at 20-$25^{\circ}C$ for 3 days in a growth medium, and the diluted solution of the microbial culture was applied under the field conditions after inoculation of the above turfgrass pathogens. The treated turfgrass was incubated at 28$^{\circ}C$ in a growth chamber. In this experiment, Pythium blight was almost completely controlled and brown patch was slightly decreased by the microbial product, while no control was observed in large patch of zoysiagrass. In zoysiagrass treated with the microbial culture, thatch accumulation was notably reduced.

• 보존과학연구
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• 통권5호
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• pp.148-166
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• 1984

The Investigation of organisms damaged to papers and cellulose materials of Cultural Property in the Ch'ang Dok Palace The investigation of the airborne fungi, the attached fungi to the papers and cellulose materials of Cultural Properties in addition to the insects inhabiting at the Ku Sonwon Jon, Shin Sonwon Jon and Yonwa ch'anggo in the Ch'ang Dok Palace carried out from Jul. 10 to Jul. 21,1984.The results are summarized as follows ;1. Isolation and identification of the airborne fungi from the three storages were Cladosporium sp., Alternaria alternata, As pergillus cervinus, A. flavus, A. nidulance, A. oryzae, A. terreus, A. versicolor, A. wentii, Penicillium adametzii, P.albicans, P.canescens, P. citreo-viride, P. citrinums, P. decumbens, P. frequentans, P. funiculosum, P.herquei, P.implicatum, P.multicolor, P.nigricans, P.nonatum,P.purpurogenum, P.roqueforti, P.viridicatum, Trichodema viride, Geotrichumcandidum, Curvvlaria lunata, Torula hebarum, T.thermophila, Itersoniliasalmonicolor, Drechsclera avenue, Candida sp., Acremonium sp., and Botrytis sp., It was found that thirty five species in thirteen genera was isolated. Among them, the dominant species was Cladosporium sp., and the order was Penicillium, Aspergillus, Alternaria and so on.2. The attached fungi directly isolated from the papers and cellulose materials of Cultural Properties were twenty-nine species in fourteen genera, namely, Acremonium sp., Albertiniella sp., Alternaria alternata, Aspergillus clavatus,A.niger, A.ornatus, A.versicolor, Botrytis sp., Bysochlamys sp., Carpenteles sp.,Chaetomium globosum, Cladosporium sp., Eurotium sp., Mucor sp., Penicilliumcanescens, P.chermesium, P.citrinum, P.frequentans, P.funiculosum, P.herqueiP .implicatum, P.javanicum, P.luteum, P.purpurogenum, P.thomii, P.viridicatum, Torula thermophila, Trichoderma koningi and T.viride. Among them, the mostfungi distributed on the surface of the papers and cellulose materials was Penicillium and the order was Aspergillus, Alternaria, Cladosporium, Trichodermaand so on.3. The insects collected the three storages were ten genera and ten species including 916 specimens. By classifying the insects collected, the most species of the insects was Stenoscelodes hayashii of 857 specimens occupied about 93% of the total insect. And the other insects were collected as Microgamme costipennisAnobium pertinax, Xenomimetes alni, Anthrenus verbasci, Holoparmecus signatus,Thermobia domestica, Halyomorpha brevis, Drosophila coracine and Brattaorientalis. As described above, it could be known that the most airborne fungi was Cladosporium and the order was Penicillium, Aspergillus, Alternaria in the three storages. And the most attached fungi distributed on the surface of papers and cellulose materials was Penicillium and the other fungi were Aspergillus, Alternaria Cladosporium, Trichoderma and so on. Accordingly, from the results, itwas assumed that the major part of airborne fungi were attached to the papers and cellulose materials of Cultural Properties. The paper and cellulose materials of Cultural Properties in Ch'ang Dok Palace were chiefly damaged by S.hayashii in Coleoptera.

• 미생물학회지
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• 제41권1호
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• pp.81-86
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• 2005

국내에서 분리된 우수섬유소분해 균주인 Trichodema sp. C-4가 생성하는 endoglucanase 중하나를 $(NH_4)_2SO_4$ 침전, Sephacryl S-200 gel filtration, DEAE-Sepharose A-50 ion exchange, Mono-P chromatofocusing (EPLC)의 단계로 정제하고 이를 F-I-III라 명명하였다. 분리된 효소 F-I-III는 분자량 56,000Da, 둥전점 4.9로 측정된 단일 단백질이었다. F-I-III는 $55^{\circ}C$에서 가장 높은 활성을 보였으며, pH 5.0이 반응 최적 조건이었다. $50^{\circ}C$에서 24시간 동안 안정하였으며, pH 4-7의 범위에서 안정하였다. CMC에 대한 비활성은 315.4U/mg 이었으며, PNPG2에 대한 Km 값은 2.69 mM이었다. 이 효소는 같은 균주에서 분리한 다른 endoglucanase와 exoglucanase를 섞었을 때 결정형 섬유소인 Avicel분해에 대한 상승효과를 보였다. $Mg^{2+},\;CO^{2+},\;Fe^{2+},\;Ca^{2+},\;CS^+,\;Li^+$ 등의 이온은 1 mM의 농도에서 효소의 활성에 큰 영향을 미치지 않았고, 1 mM의 환원제 (cystein, EDIA, \beta-mercaptoethanol, dithiothreitol(DTT), L-ascorbic acid)들은 효소의 활성을 증가시켰다. E-I-III의 N-말단 서열을 분석하여 QPGTSTPEVHPKKLTTYK의 서열을 얻었다. 이는 Trichodema reesei의 endoglucanase인 EGI과 $95\%$의 유사도를나타내었다. 분리된 효소 F-I-III는 높은 비활성을 가지고 있어서 활용가치가 높을 것으로 사료되었다.

• 보존과학연구
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• 통권17호
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• pp.48-64
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• 1996

We compared the degree of color difference formed by environmental factor(temperature, relative humidity) with fungal growth in order to know how to change the color difference of cellulolutic cultural properties such as Korean papers, cotton, jute and hemp. We concluded, from the result, that the action of fungal growth on celluloytic cultural properties was more hamful than environmental factor. We considered the secretion produced by fungi as the causative agent for stained formation on cellulolytic cultural properties. Alternaria sp. colored allmaterials greyish black, Chaetomium sp. colored cotton and hemp orange, and Penicllium sp. colored cotton, jute and hemp yellowish green. But Trichoderma sp. and Aspergillus sp. didn't show a clear color against each material. It was observed that thymol(120g/$m^3$) was the most effective fungicide to prevent fungal growth.

• 환경생물
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• 제20권2호
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• pp.130-135
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• 2002

PDA배지에서 $25^{\circ}C$로 배양할 때 초기 균사의 색은 흰색을 띄고 있었으며 배양 후기에 생성되는 포자의 색은 짙은 푸른색을 나타내고 있었다. 시간당 성장률은 1.1mm보다 빠른 것으로 나타났으며 성숙한 포자의 모양은 부드럽고 구형에 가까운 모양이었다. 이러한 형태학적 결과는 시간당 성장률이 0.5-0.7mm 이하인 Tricho-derma viride 및 T. hamatum의 특징과 성숙 포자의 모양이 실린더 모양인 T. koningii, T longibrachiatum과 구별되는 결과였다(Raifai 1969; Seaby 1996; Gams and Bissett 1998). 또한 Trichoderma harzianum의 분류에 있어 가장 중요한 구조는 phialospore의 형태가 구형이어야 하며, 포자의 겉은 부드럽고 크기는 $2.5-3.0\times{3.4-4.0}\mu{m}$ 정도인 것이 특징이다(Morris and Doyle 1995a, b). 이러한 결과를 종합하여 분리 균주를 동정한 결과 Trichoderma harzianum으로 확인할 수 있었다. Phytophthora capsici에 대한 분리 균주의 성장저해정도를 측정한 결과 $27^\circ{C}$의 조건에서 가장 높은 길항력을 나타내었으며 다른 곰팡이에 대한 성장억제도 이 온도에서 가장 높을 것으로 판단되었다. 또한 $31^\circ{C}$ 이상에서는 T. harzianum SJG-99721의 성장 속도가 급속히 떨어짐을 확인한 바 이러한 성질을 이용하여 느타리버섯의 균사 배양시 고온 배양을 실시할 경우 푸른곰팡이 병 오염을 극히 제한할 수 있을 것으로 판단되었다. 또한 이러한 고온에서의 성장 속도 제한은 식물 병원성 미생물과의 길항 작용에서도 영향을 미치는 만큼 생물농약으로서의 사용은 토양 온도가 $30^\circ{C}$ 넘지 않는 기간에 최대한 토양 내에서 증식이 이루어지도록 조처해야 한다. 진균성 식물 병원균의 세포벽 분해효소인 extracellular chitinase의 높은 활성도를 확인함으로서 Trichoderma harzianum SJG-99721의 강력한 식물 병원균 제어능력은 이들의 높은 chitinase의 활성도에 의한 것으로 유추할 수 있었다.

• 한국버섯학회지
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• 제9권3호
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• pp.110-115
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• 2011

느타리버섯 푸른곰팡이병에 대한 품종 저항성은 포장에서 효과적인 검토가 불가능하여 한천 배지상에서 푸른곰팡이균이 균을 저해하는 특징 fungistatic, lysis, toxin enzyme 등에 대한 각각의 특성을 실내에 검정할 수있는 방법을 검토하기 위하여 균사생장과 특이 현상을 조사하였다. 대치배양에서는 느타리버섯 균주에 따라 대치선 형성위치, 푸른곰팡이균의 느타리균사 생장부분 위로 생장(overgrowth), 대치선 이후의 버섯균의 용균(lysis)현상이 발생하여 효과적인 저항성 검정이 가능하였다. 배양여액을 여지에 적용하는 방법은 버섯균과 병원균의 종류에 따른 균사생장 및 특이적 현상이 타나나지 않았으나, well plate를 이용한 배양여액 희석방법으로 균사생장의 억제정도의 확인이 가능하였다. 분활 petri dish를 활용하여 동시배양의 경우 약간의 버섯균이 억제되는 현상은 보였으나, 푸른곰팡이균이 버섯균 생장 부분 위로 덮어 효과검정이 불가능 하였다. 버섯균을 petri dish 전체에 배양한 후 그 위에 병원균의 균총을 접종하는 방법은 overgrowth, lysis등의 현상이 발생하여 병원성의 검정이 용이하였다. 실내에서의 느타리버섯균의 푸른곰팡이병에 대한 저항성 검정은 대치배양, 배양여액법, 생장후 접종법에 의한 방법으로 가능한 것으로 판단되었다.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2009년도 춘계학술대회 및 임시총회
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• pp.38-40
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• 2009

• 한국균학회소식:학술대회논문집
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• 한국균학회 2014년도 춘계학술대회 및 임시총회
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• pp.41-41
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• 2014

Pinewood nematode (PWN, Bursaphelenchus xylophilus) is a serious pathogenic worm that quickly dry pine trees to death. Recently, PWN has been devastating huge amounts of conifer trees in Korea. As a first step to explore the association and ecological roles of fungi in PWN life cycle in Korea, in this study we first isolated and indentified fungi from PWN-infested Korean pine and Japanese black pine wood sampled in Jinju, Sacheon, Pocheon, Chuncheon, Gwangju, and Hoengseong in Korea. A total of 144 fungal isolates were obtained from Japanese black pine wood and 264 fungal isolates from Korean pine wood. Their morphology and nucleotide sequences of the ITS rDNA and ♌-tubulin gene were examined for species identification. Ophiostoma ips, Botrytis anthophila, Penicillium sp., Hypocrea lixii, Trichoderma atroviride, O. galeiforme, Fusarium proliferatum were identified from Japanese black pine wood. Leptographium koreanum, L. pini-densiflorae, Ophiostoma ips, Penicillium raistrick, Trichoderma sp. were isolated from Korean pine wood. O. ips and L. koreanum were the major species on the two different PWN-infected pine tree. The cultivation of PWN on fungal mat of the identified species did some enhance PWN reproduction. The ambrosia beetle, Platypus koryoensis, is a serious pest of oak trees in Korea. In this study we investigated filamentous fungi present in the body of the beetle. Fourteen genera of filamentous fungi belonging to Ascomycota and Basidiomycota were isolated. All the obtained genera were isolated in the mitosporic state. The identified fungi were classified in 11 distinct orders including the Ascomycota (Eurotiales, Hypocreales, Microascales, Ophiostomatales, Pleosporales, and Sordiales) and Basidiomycota (Agaricales, Corticiales, Polyporales, and Russulales Xylariales). Within Ascomycota, 13 species were found. Meanwhile five species were found within Basidiomycota. The results showed the presence of diverse fungi in P. koryoensis. Among the isolated fungi, some were able to produce wood degrading enzymes. Further fungal isolation was performed with P. koryoensis infested Quercus mongolica trees sampled at Kumdan mountain in Hanam-Si, Gyeonggi province from June of 2009 to June of 2010. Penicillin spp. and Trichoderma spp. were the major species of mold fungi group. Pichia guilliermondii was the major species of mold yeast group. Raffaelea quercus-mongolicae was also isolated, but its isolation frequency was not high. Other species identified were Ambrosiella xylebori, Fusarium solani, Cryphonectria nitschke, Chaetomium globosum, and Gliocladium viride, Candida kashinagacola, C. maritima, C. vanderkliftii, Saccharomycopsis crataegensis.