• 미생물학회지
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• 제42권2호
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• pp.135-141
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• 2006

덩굴마름병균인 Didymella bryoniae KACC 40669에 대하여 높은 항진균 활성을 나타내는 Brevibacillus sp. KMU-391은 Russia의 Sanktpeterburg 지역의 토양 시료로부터 분리하였다. Brevibacillus sp. KMU-391이 생산하는 항진균 물질은 D. bryoniae의 생육을 강하게 억제할 수 있었으며 TSB를 기본배지로 하여 탄소원으로 1.0% sucrose와 1.0%의 polypeptone을 첨가한 후 pH를 7.0으로 조정하여 $30^{\circ}C$에서 180 rpm으로 2일간 배양하였을 때 가장 높은 항진균 활성을 나타내었다. 배양액에 $30{\sim}60%$의 ammonium sulfate를 처리하여 Brevibacillus sp. KMU-391이 생산하는 항진균 물질을 회수한 후 D. bryoniae를 액체배양을 통한 항진균 활성을 조사한 결과 대조구와 비교하여 41%의 생육을 저해하는 것으로 확인되었다. 또한 butanol을 이용하여 항잔균 물질을 회수한 후, 다양한 작물병원성 곰팡이를 대상으로 항진균 활성 spectrum을 조사한 결과 조사, 장미 잿빛곰팡이병, 잠두 붉은점무늬병, 고추 탄저병, 참외 탄저병, 수박 등의 박과작물의 덩굴마름병, 보리 질병, 토마토 시들음병, 글라디올러스 마른썩음병, 토마토 질병, 수박 덩굴쪼김병, 수박 질병, 사과나무 역병, 벼 잎집무늬마름병, 참외 줄기썩음병, 그리고 고추균핵병에 대하여 높은 항진균 활성을 나타내었다. 이러한 결과를 바탕으로 Brevibacillus sp. KMU-391을 이용하여 작물병원성 곰팡이에 대한 생물농약으로서의 응용 가능성을 확인 할 수 있었다.

• 한국식물병리학회지
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• 제13권2호
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• pp.105-112
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• 1997

Didymella bryoniae, gummy stem blight fungus of cucurbits, has been known not to produce its pycnidium in vitro without irradiation. Various methods for producing pycnidiospores of the fungus as an inoculum have been used. However, those methods have not been verified in terms of efficiency of the productivity, activity and synchronous maturation of the inoculum. Therefore, a pycnidiospore production method in vitro that is highly reliable and reproducible has to be developed to obtain a large amount of inoculum for screening disease resistant varieties or effective fungicides. Here we standardized a mass-production technique for pycnidiospores of D. bryoniae in vitro by comprehensively finding the optimal conditions such as kinds and thickness of cultural medium, growing temperature, and quality and duration of irradiation as well as examining the activity and pathogenicity of the pycnidiospores reproduced. In brief, mycelial colony on the PDA plate was cultured at 26$^{\circ}C$ for 2 days under the darkness, and then the plate was irradiated under the UV light (12 hr/a day) for 2~3 days at the same temperature(26$^{\circ}C$). Two days after UV irradiation, a great number of pycnidia was simultaneously formed. This plate was subjected to darkness again for 4~5 days to mature pycnidiospores. We could obtain a large amount of inoculum that is synchronously matured in a short period of time through the above procedures.

• Mycobiology
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• 제38권3호
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• pp.166-170
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• 2010

Gummy stem blight is a major foliar disease of muskmelon (Cucumis melo L.). In this study, morphological characteristics and rDNA internal transcribed spacer (ITS) sequences were analyzed to identify the causal organism of this disease. Morphological examination of the Jeonbuk isolate revealed that the percentage of monoseptal conidia ranged from 0% to 10%, and the average length $\times$ width of the conidia was 70 ($\pm$ 0.96) $\times$ 32.0 ($\pm$ 0.15) ${\mu}m$ on potato dextrose agar. The BLAST analysis showed nucleotide gaps of 1/494, 2/492, and 1/478 with identities of 485/492 (98%), 492/494 (99%), 491/494 (99%), and 476/478 (99%). The similarity in sequence identity between the rDNA ITS region of the Jeonbuk isolate and other Didymella bryoniae from BLAST searches of GenBank was 100% and was 95.0% within the group. Nucleotide sequences of the rDNA ITS region from pure culture ranged from 98.2% to 99.8%. Phylogenetic analysis with related species of D. bryoniae revealed that D. bryoniae is a monophyletic group distinguishable from other Didymella spp., including Ascochyta pinodes, Mycosphaerella pinodes, M. zeae-maydis, D. pinodes, D. applanata, D. exigua, D. rabiei, D. lentis, D. fabae, and D. vitalbina. Phylogenetic analysis, based on rDNA ITS sequence, clearly distinguished D. bryoniae and Didymella spp. from the 10 other species studied. This study identified the Jeonbuk isolate to be D. bryoniae.

• 식물병과 농업
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• 제5권1호
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• pp.20-26
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• 1999

Ultraviolet light is required for the sporulation of Didymella bryoniae, a gummy stem blight fungus in cucurbits such as watermelon, melon, oriental melon, cucumber and pumpkin. In this experiment, the upper limit of wave length for the production of pycnidia of D. bryoniae was 365 nm - 375 nm. Two plastic houses were covered with either common transparent film (wave length longer than 225 nm is transmitted) or UV-absorbing film ( wave lenght shorter than 388 nm is absorbed). In both houses, seedlings inoculated with D. bryoniae were placed in the center of the house at 30 days after transplantation of watermelon (cv. Whanhoseong), and the disease incidences between the houses were compared until 80 days after transplantation. The number of disease lesions and incidence of pycnidia-producing lesions under the UV-absorbing film were reduced by 90% and 80%, respectively, compared to the common transparent film. The internode lengths of plants grown in the two houses were not significantly different, but the plants grown under the UV-absorbing film had longer vines and more leaves than plants under the common transparent film. However, fruit characters such as weight, length, width, rind thick and brix, were not different between the two houses. Occurrence of aphids was reduced in the UV-absorbing film, but those of mites or diseases (powdery mildew and sooty mold) were not different between the houses. These results suggest that disease incidence of gummy stem blight of watermelon in the greenhouse can be controlled by the use of UV-absorbing film.

• 식물병연구
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• 제10권4호
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• pp.313-318
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• 2004

우리나라에서 오이 속썩음은 대전을 비롯한 여러 오이 재배지역에서 발생하였다. 본 병의 발생은 조사포장에서 평균 4.2%, 최대 21.5%까지 발생하였다. 병 발생은 오이 꽃이 붙어 있는 부위부터 시작되었다. 병원균을 접종하여 감염된 오이의 내부조직은 길이 2cm, 폭이 2mm 이상으로 갈변되었다. 후에 갈변은 진전되어 과일의 심피에 이르렀으며, 감염된 오이의 끝부분이 울퉁불퉁하게 되었다. 병반부위에서 분리된 병원균은 형태적 특징에 의하여 Didymella bryoniae으로 동정되었다. 병원균의 균사생장 온도범위는 $5{\sim}32^{\circ}C$이고, 최적 균사생장 온도는 $26{\sim}28^{\circ}C$이었다. 포자현탁액을 오이 꽃에 접종하여 오이 과일의 내부에서 자연감염된 것과 유사한 병징이 나타났다. 오이, 수박, 참외, 멜론 및 호박에서 분리된 D. bryoniae균주들은 접종실험에 의하여 오이 속썩음이 나타났다. 오이 속썩음의 발생 온도범위는 $10{\sim}32^{\circ}C$이며, 최적 발생온도는 $25{\sim}28^{\circ}C$이었다.

• 한국미생물·생명공학회지
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• 제32권3호
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• pp.238-242
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• 2004

D. bryoniae를 원인균으로 하는 참외 만고병에 대해서 항만고병 활성물질을 생산하는 미생물을 방선균으로부터 선별 분리한 후 배양액으로부터 항균활성물질의 규명을 시도하였다. 항진균 물질과 같은 이차대사산물의 생산에 증가시키는 $K_2$$HPO_4$와 칼슘이온이 포함된 GSS배지에서 방선균 SKM338 균주를 180 rpm, $30^{\circ}C$, 5일 동안 배양하여 얻어진 배양 상등액으로부터 물리화학적인 방법으로 항진균 활성이 있는 물질을 분리 정제한 결과 참외의 만고병에 대한 생물농약으로 개발 가능한 방선균 유래의 항진균성 물질은 NMR, IR, UV 및 Mass spectral data 분석 등을 통해 polyene macrolide계에 속하는 항생물질인 Flavofungin, Fungichromin, Filipins로 밝혀졌으며 이들의 응용을 기대해 본다.

• 한국식물병리학회지
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• 제1권3호
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• pp.173-177
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• 1985

본 연구는 자연이병된 오이와 호박 종자에서 분리한 덩굴마름병균(만고병균)의 병원성의 차이를 조사한 것이다. 오이, 참외, 호박 및 수박의 유묘에 대한 일차 감염은 유근$\cdot$배축$\cdot$자엽에 나타났으며 병징은 작물에 관계없이 비슷하였다. 유근이 병들었을때는 발아전 부패현상으로 나타났고 배축과 자엽의 감염은 제1엽과 줄기로 이어지는 접종원이었다. 교호접종시험 결과 공시된 덩굴마름병균의 모든 분리균은 오이, 참외, 호박, 수박의 종자 및 유묘에 대하여 병원성이 있었으나 분리균이나 공시작물간에는 차이가 별로 나타나지 않았다. 오이와 호박의 감수성은 다습조건에 의해서 크게 영향을 받았다.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.133.1-133
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• 2003

Gummy stem blight, caused by Didymella bryoniae occurs exclusively on cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. Through this study, we optimized cultural conditions for mass-production of pycnidiospore by Metal Halide Lamp irradiation. In brief, the mycelial was cultured at $26^{\circ}C$ on PDA, for 2 days under the darkness, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$, a great number of pycnidia was simultaneously formed. Thus produced pycnidiospores were used as immunogen. From fusions of myeloma cell (v-653) with splenocytes from immunifed mice were car ried out. And, two hybridoma cell lines that recognized the immunogen Didymella bryoniae were obtained. One Monoclonal Antibody, Db1, recognized the supernatant and the other monoclonal antibody, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid two MAb Db1 and Db15, were immunotyped and identified as IgG1 and IgG2b, respectively. Titer of MAb Db1 and MAb Db15 was measured absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with Didymella bryoniae but none reacted with other of fungi and CMV, CGMMV Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{3}$ well by indirect ELISA characterization of the MAb Db1, Db15 antigen by heat and protease treatments show that the epitope recognized by the MAb Bb1, Db15 were a glycoprotein.

• 한국응용곤충학회지
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• 제16권4호
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• pp.211-215
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• 1977

오이류 덩굴마름병균(Didymella bryoniae)의 포자형성에 미치는 배지, 근자외선광과 화광색형광등 12시간 교호조사와 계속암흑처리의 영향을 조사하였다. Didymella bryoniae를 감자한천과 V-8 juice 한천에 배양하였을 때 광선은 병자각 및 자양각의 형성을 촉진하였으나 계속 암흑처리구의 포자형성은 낮았다. 그러나 균주에 따라서는 무광하에서도 병자각과 자양각을 형성할 수 있었다. V-8 juice 한천에 배양했을 때 자양각의 형성은 근자외선 보다는 화광색광선에서 촉진되었다. 일반적으로 광원에 관계없이 V-8 juice 한천에 배양한 것이 감자한천에 배양한 것보다 포자의 형성이 양호하였다. 감자한천에서 배양된 C. bryoniae의 각균주는 병포자의 대부분이 무격막의 소형 병포자였음에 비하여 V-8 juice 한천에 배양한 것은 무격막의 대형 병포자 또는 1개의 격막을 가진 병포자로써 병든 종자의 배축상에 형성된 배포자와 흡사하였다. 자양각의 외관은 병자각과는 판이하였으며 유두상 각공위에 백색의 포자괴가 형성되었다.

• The Plant Pathology Journal
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• 제22권1호
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• pp.36-45
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• 2006

Twenty isolates of Didymella bryoniae were isolated from infected cucurbit plants in various growing areas of southern Korea in 2001 and 2002. Random Amplified Polymorphic DNA (RAPD) group [RG] I of D. bryoniae was more virulent than RG IV to watermelon. Virulence of the RG I isolate was strong to moderate to cucumber, whereas that of the RG IV varied from strong, moderate to weak. Two hundred seventy-three amplified fragments were produced with 40 primers, and were analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYSPC. At the distance level of 0.7, two major genomic DNA RAPD groups were differentiated among 20 isolates. The RG I included 7 isolates from watermelon and one isolate from melon, whereas the RG IV included 12 isolates from squash, cucumber, watermelon and melon. Amplification of internal transcribed spacer (ITS) region and small subunit rRNA region from the 20 isolates yielded respectively a single fragment. Restriction pattern with 12 restriction enzymes was identical for all isolates tested, suggesting that variation in the ITS and small subunit within the D. bryoniae were low. Amplification of the genomic DNAs of the tested isolates with the sequence characterized amplified regions (SCAR) primer RG IF-RG IR specific for RG I group resulted in a single band of 650bp fragment for 8 isolates out of the 20 isolates. Therefore, these 8 isolates could be assigned into RG I. The same experiments done with RG IIF-RG IIR resulted in no amplified PCR product for the 20 isolates tested. An about 1.4 kb-fragment amplified from the RG IV isolates was specifically hybridized with PCR fragments amplified from genomic DNAs of the RG IV isolates only, suggesting that this PCR product could be used for discriminating the RG IV isolates from the RG I isolates as well other fungal species.