• Title/Summary/Keyword: gummy stem blight

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Population Structure of Stagonosporopsis Species Associated with Cucurbit Gummy Stem Blight in Korea

  • Jeong, Yong-Jik;Kwon, Oh-Kyu;Jeong, A-Ram;Lee, Hyunji;Moon, Hyeran;Lee, O New;Hong, Jeum Kyu;Park, Chang-Jin
    • The Plant Pathology Journal
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    • v.38 no.5
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    • pp.522-532
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    • 2022
  • Gummy stem blight (GSB), a common and serious disease in cucurbits worldwide, is caused by three genetically distinct species: Stagonosporopsis cucurbitacearum (syn. Didymella bryoniae), S. citrulli, and S. caricae. In Korea, however, the three species of Stagonosporopsis have been barely characterized. In this study, 21 Stagonosporopsis isolates were recovered from watermelon (Citrullus lanatus) and muskmelon (Cucumis melo) leaves and stem showing blight symptoms collected from 43 fields in Korea. Sequence analysis performed with an internal transcribed spacer region was not competent to differentiate the Stagonosporopsis isolates. On the contrary, analysis of β-tubulin (TUB) genes and three microsatellite markers, Db01, Db05, and Db06, successfully differentiated Stagonosporopsis isolates. Further sequence analysis identified two Stagonosporopsis species, S. citrulli and S. caricae, and one previously unknown species of Stagonosporopsis. Representative isolates from three species caused dark water-soaked lesions on the detached watermelon and muskmelon leaves with no significant differences in the aggressiveness. Our results indicate that the S. citrulli, S. caricae, and unknown Stagonosporopsis sp. are all causal agents of GSB for both watermelon and muskmelon. This is the first report of a new species and the population structure of Stagonosporopsis species causing GSB in Korea.

Ultraviolet Wave Length Effective in the Sporulation of Didymella bryoniae, a Gummy Stem Blight Fungus in Cucurbits, and the Disease Control Effect by the Use of Ultraviolet Light-Absorbing Vinyl Film (박과작물 덩굴마름병권 Didymella bryoniae의 포자형성 유효 자외파장과 자외선 흡수필름을 이용한 병 방제효과)

  • 권미경;홍정래;기운계;조백호;김기청
    • Plant Disease and Agriculture
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    • v.5 no.1
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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.

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Standardization of a Mass-Production Technique for Pycnidiospores of Dydymella bryoniae, Gummy Stem Blight Fungus of Cucurbits (박과작물 덩굴마름병 Didymella bryoniae의 병포자 대량 생산 방법의 표준화)

  • 권미경;홍정래;선해정;성기영;조백호;김기청
    • Korean Journal Plant Pathology
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    • v.13 no.2
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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.

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Diseases Occurrence rind Fermented Fruit Development by Culture Methods and Rootstocks of Oriental Melon(Cucumis mezo L.) (참외 재배법 및 대목 종류에 따른 병해 및 발효과 발생)

  • 연일권;신용습;배수곤;도한우;박종욱;박소득
    • Journal of Bio-Environment Control
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    • v.11 no.1
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    • pp.18-22
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    • 2002
  • Diseases occurrence of hydroponic culture was less than that of soil culture, especially gummy stem blight, which was 39.9-53.3% of infection rate at soil culture, was not almost observed at hydroponic culture. Development of fermented fruit was higher at Shintozoa, Hongtozoa and self-rooting seedling in order at soil cultured but critically dropped at hydroponic culture without rootstock sort. Therefore proper moisture management at root zone can decrease the development of fermented fruit.

Identification and Characterization of the Causal Organism of Gummy Stem Blight in the Muskmelon (Cucumis melo L.)

  • Choi, In-Young;Choi, Jang-Nam;Choi, Dong-Chil;Sharma, Praveen Kumar;Lee, Wang-Hyu
    • Mycobiology
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    • v.38 no.3
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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.

Screening of melon genotypes identifies gummy stem blight resistance associated with Gsb1 resistant loci

  • Hassan, Md Zahid;Robin, Arif Hasan Khan;Rahim, Md Abdur;Natarajan, Sathishkumar;Kim, Hoy-Taek;Park, Jong-In;Nou, Ill-Sup
    • Journal of Plant Biotechnology
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    • v.45 no.3
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    • pp.217-227
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    • 2018
  • Gummy stem blight (GSB) is one of the most destructive and economically important, soil borne diseases of melon caused by the ascomycete fungus, Didymella bryoniae throughout the world. In Korea, however, no GSB resistant genotype has been reported yet. The study aimed to identify GSB resistant melon germplasm. We screened a total of 60 genotypes including 16 lines and 44 melon cultivars collected from USA and Korea. Among the 16 melon lines, four lines including 'PI482399', 'PI140471', 'PI136170' and 'PI420145', and two Korean cultivars viz. 'Asia Papaya' and 'Supra' showed complete resistance. We were aware that both genotypic and environmental variations could influence the phenotypic screening of resistance and susceptibility. We therefore, further assessed all genotypes using 20 SSR markers. The SSR marker 'CMCT505' linked to Gsb1 in chromosome 1 perfectly grouped resistant and susceptible lines indicating that resistance is probably due to the presence of Gsb1 gene. Cloning and sequencing of resistant and susceptible Gsb1 amplicons showed that there were 32-bp deletions in resistant line and 39-bp deletions in resistant cultivar compared to susceptible one. Thus, the resistant melon lines and cultivars identified in this study could be recommended for the melon breeding program. Furthermore, the SSR marker 'CMCT505' which is tightly linked with Gsb1 could be used for molecular screening of melon germplasm.

Isolation of Polyene Antifungal Antibiotics Against Gummy Stem Light Caused by Didymella bryoniae (Streptomyces sp. 유래 Polyene 계 항만고병 항생물질의 분리)

  • 김광석;서영배
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.238-242
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    • 2004
  • Antifungal agents, flavofungin and fungichromin were isolated from the fermentation culture broth of a Streptomyces sp. SKM338. Biological evaluation of these antibiotics indicated that the compounds possesses broad spectrum antifungal activity against various pathogens. Especially, these compounds inhibited throughly growth of Didymella bryoniae, caused Gummy stem blight of melons, occurs in the southeastern Korea. Inhibition of this pathogen may be prevented from directly reducing both pre- and post-harvest yields.

Screening of Resistant Watermelon Cultivars Against Gummy Stem Blight Fungus, Didymella bryoniae, and Comparison of Protein Expression Between Cultivars After Infection (수박 덩굴마름병의 품종 저항성 검정과 감염 후 품종간 단백질 발현의 비교)

  • 홍정래;임양주;권미경;조백호;김기청
    • Korean Journal Plant Pathology
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    • v.14 no.4
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    • pp.339-344
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    • 1998
  • Since the leaf inoculation procedures are time-consuming and require considerable growth chamber space, a rapid dioassay method for screening of pathogenicity of Didymella bryoniae, a casual agent of gummy stem blight in watermelon, was established in this paper. The method produced reliable results within 8 days ( 5 days for growing seedlings and 3 days for rapid disease response in the seedlings). After contaminants in the root of 4~5 day-old seedlings had been washed using sterilized water, 5 seedlings were dipped into a vial containing 12 ml of conidial suspension (106 cells/ml). After the vials were placed in a growth chamber (22$^{\circ}C$, RH 50%, 14hr light/10hr darkness) for 3 days, susceptibility and resistance of cultivars were determined by the degree of disease response on cotyledon. The result of obtained by the dip-inoculation method was well coincided with the results by the leaf inoculation procedures and the result that had been observed for several years in the field. Screening of collected watermelon cultivars by the dip-inoculation method revealed that all the 21 domestic cultivars collected were susceptible and only 3 foreign cultivars (PI 189225, PI 482322 and IT 188207) were resistant among 18 cultivars A cucumber cultivar (Marketer) and bitter cucumber were proven to be resistant against the D. bryoniae among 8 other different cucurbits tested. The SDS-PAGE patterns of total proteins from a susceptible (Keumcheon) and a resistant (PI 189225) watermelon cultivars were compared 0, 12, 24 and 36 hrs after inoculation. The amounts of two distinct protein bands (24 kDa and 70 kDa) were gradually increased after inoculation in both cultivars.

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Monoclonal Antibody-Based Indirect-ELISA for Early Detection and Diagnosis of Epiphytic Didymella bryoniae in Cucurbits

  • Lee, Sun-Cheol;Han, Ki-Soo;Lee, Jung-Han;Kim, Dong-Kil;Kim, Hee-Kyu
    • The Plant Pathology Journal
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    • v.19 no.5
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    • pp.260-265
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    • 2003
  • Gummy stem blight caused by Didymella bryoniae occurs exclusively in cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. In this study, cultural conditions for the mass-production of pycnidiospore by Metal Halide (MH) lamp irradiation were maximized. The mycelia were cultured at $26^{\circ}C$ on PDA for 2 days under dark condition, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$. Results show that a great number of pycnidia were simultaneously formed. The pycnidiospores produced were then used as immunogen. Fusions of myeloma cell (v-653) with splenocytes from immunized mice were carried out. Two hybridoma cell lines that recognized the immunogen D. bryoniae were obtained. One monoclonal antibody (MAb), Dbl, recognized the supernatant while another MAb, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid of the two MAb, Dbl and Db15, the immunotypes of which were identified as IgG1 and IgG2b, respectively. Titers of MAb Dbl and MAb Db15 were measured and the absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with D. bryoniae but none reacted with other viral isolates, Cucumber mosaic virus and Cucumber green mottle mosaic virus. Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{-3}$/well by indirect ELISA. Characterization of the MAbs Dbl, Db15 antigen by heat and protease treatments, which suggests that the epitope recognized by these two MAbs was glycoprotein.