• Title/Summary/Keyword: Wilt pathogen

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Detection and Quantification of Fusarium oxysporum f. sp. niveum Race 1 in Plants and Soil by Real-time PCR

  • Zhong, Xin;Yang, Yang;Zhao, Jing;Gong, Binbin;Li, Jingrui;Wu, Xiaolei;Gao, Hongbo;Lu, Guiyun
    • The Plant Pathology Journal
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    • v.38 no.3
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    • pp.229-238
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    • 2022
  • Fusarium wilt caused by Fusarium oxysporum f. sp. niveum (Fon) is the most serious soil-borne disease in the world and has become the main limiting factor of watermelon production. Reliable and quick detection and quantification of Fon are essential in the early stages of infection for control of watermelon Fusarium wilt. Traditional detection and identification tests are laborious and cannot efficiently quantify Fon isolates. In this work, a real-time polymerase chain reaction (PCR) assay has been described to accurately identify and quantify Fon in watermelon plants and soil. The FONRT-18 specific primer set which was designed based on identified specific sequence amplified a specific 172 bp band from Fon and no amplification from the other formae speciales of Fusarium oxysporum tested. The detection limits with primers were 1.26 pg/µl genomic DNA of Fon, 0.2 pg/ng total plant DNA in inoculated plant, and 50 conidia/g soil. The PCR assay could also evaluate the relationships between the disease index and Fon DNA quantity in watermelon plants and soil. The assay was further used to estimate the Fon content in soil after disinfection with CaCN2. The real-time PCR method is rapid, accurate and reliable for monitoring and quantification analysis of Fon in watermelon plants and soil. It can be applied to the study of disease diagnosis, plant-pathogen interactions, and effective management.

First Report of Charcoal Rot Caused by Macrophomina phaseolina on Peanut Plants in Korea (땅콩에서 Macrophomina phaseolina에 의한 균핵마름병 발생 보고)

  • Soo Yeon Choi;You Kyoung Lee;Chang Ok Geum;Shinhwa Kim;Hyunjung Chung;Sang-Min Kim;Yong Hoon Lee
    • The Korean Journal of Mycology
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    • v.51 no.4
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    • pp.383-387
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    • 2023
  • Peanut plants showing mild wilt were found in fields of Iksan, Korea, in August 2021. The diseased peanut plants were collected, and the causal pathogens were isolated using potato dextrose agar (PDA) medium. The isolated IS-1 strain formed white mycelia on PDA, which turned black with age. Sclerotia were produced on the PDA and barley leaves laid on water agar 7 d after incubation at 30℃. The sequences of both the internal transcribed spacer (ITS) region and calmodulin gene of IS-1 showed a 100% similarity with that of Macrophomina phaseolina. A phylogenetic tree constructed using the ITS regions of fungal pathogens causing disease in peanut plants indicated that the IS-1 stain belongs to M. phaseolina. The inoculation of IS-1 sclerotia into peanut seedlings resulted in yellowing and wilt symptoms in aboveground plants and brown to dark rots in roots 35-40 d after inoculation. Overall, the morphological characteristics, molecular identification, and pathogenicity of IS-1 indicate that the causal pathogen is M. phaseolina. This is the first report of charcoal rot caused by M. phaseolina on peanut plants in Korea. Further study is needed to develop the control measures for charcoal rot in peanut plants.

Development of W/O/W Multiple Emulsion Formulation Containing Burkholderia gladioli

  • KIM, HWA-JIN;CHO, YOUNG-HEE;BAE, EUN-KYUNG;SHIN, TAEK-SU;CHOI, SUNG-WON;CHOI, KEE-HYUN;PARK, JI-YONG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.29-34
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    • 2005
  • W/O/W (water-in-oil-in-water) type multiple emulsion was applied to improve the storage stability of an antagonistic microorganism, Burkholderia gladioli. Encapsulation of microorganism into a W/O/W emulsion was conducted by using a two-step emulsification method. W/O/W emulsion was prepared by the incorporation of B. gladioli into rapeseed oil and the addition of polyglycerin polyriconolate (PGPR) and castor oil polyoxyethylene (COG 25) as the primary and secondary emulsifier, respectively. Microcrystalline cellulose was used as an emulsion stabilizer. To evaluate the usefulness of W/O/W emulsion formulation as a microbial pesticide for controlling the bacterial wilt pathogen (Ralstonia solanacearum), the storage stability and antagonistic activity of emulsion formulation were tested in vitro. The storage stability test revealed that the viability of formulated cells in emulsion was higher than that of unformulated cells in culture broth. At $4^{\circ}C$, the viabilities of formulated cells and unformulated cells at the end of 20 weeks decreased to about 2 and 5 log cycles, respectively. At $37^{\circ}C$, the viability of formulated cells decreased to only 2 log cycles at the end of storage. On the other hand, the viable cells in culture broth were not detected after 13 weeks. In activity test, formulated cells in emulsion were more effective in inhibiting the growth of pathogen than unformulated cells in culture broth. Unformulated cells completely lost their antagonistic activity during storage under similar conditions. The W/O/W multiple emulsion formulation was shown to be useful as the novel liquid formulation for biological control.

Macromolecular Docking Simulation to Identify Binding Site of FGB1 for Antifungal Compounds

  • Soundararajan, Prabhakaran;Sakkiah, Sugunadevi;Sivanesan, Iyyakkannu;Lee, Keun-Woo;Jeong, Byoung-Ryong
    • Bulletin of the Korean Chemical Society
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    • v.32 no.10
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    • pp.3675-3681
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    • 2011
  • Fusarium oxysporum, an important pathogen that mainly causes vascular or fusarium wilt disease which leads to economic loss. Disruption of gene encoding a heterotrimeric G-protein-${\beta}$-subunit (FGB1), led to decreased intracellular cAMP levels, reduced pathogenicity, colony morphology, and germination. The plant defense protein, Nicotiana alata defensin (NaD1) displays potent antifungal activity against a variety of agronomically important filamentous fungi. In this paper, we performed a molecular modeling and docking studies to find vital amino acids which can interact with various antifungal compounds using Discovery Studio v2.5 and GRAMMX, respectively. The docking results from FGB1-NaD1 and FGB1-antifungal complexes, revealed the vital amino acids such as His64, Trp65, Ser194, Leu195, Gln237, Phe238, Val324 and Asn326, and suggested that the anidulafungin is a the good antifungal compound.The predicted interaction can greatly assist in understanding structural insights for studying the pathogen and host-component interactions.

Infection Route of Bacterial Wilt of Tomato Caused by Ralstonia. solanacearum in Hydroponic Culture (수경재배에서 토마토풋마름병의 전염경로)

  • Nam, Ki-Woong;Moon, Byung-Woo;Kim, Young-Ho;Lee, Chang-Hee
    • Journal of Bio-Environment Control
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    • v.18 no.2
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    • pp.171-176
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    • 2009
  • Hydroponic culture has been developed to control soilborn diseases, to increase yield and to enhance the quality of vegetable, The pathogen could be detected from infected plant materials, hydroponic tanks, culture solution and solid media of the severely infected greenhouse, The density of pathogen population was coincided with the severity of disease incidence, For example, 1,900cfu m$L^{-1}$ pf pathogens were counted from tomato plants sampled in a 20% diseased greenhouse. The pathogens may be introduced in the greenhouse through the contaminated soil surrounding the house and/or through the infected young seedlings grown on the nursery soil. Also, not detected to Ralstonia solanacearum from tomato seeds (House Momotaro, Bbaebbae, Ggoggo, and Minicarol cultivar) selling at a market.

Evaluation of genetic affinities among Fusarium oxysporum f. sp. fragariae by RAPD and rDNA RFLP

  • Gopal Nagarajan;Nam, Myeong-Hyeon;Lee, Yun-Soo;Lee, Woo-Chung;Yoo, Sung-Joon;Song, Jeong-Young;Kim, Hong-Gi
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.116.1-116
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    • 2003
  • Fusarium oxysporum f. sp. fragariae is a fungal pathogen causing wilt disease on strawberry. The RAPD and RFLP of IGS region of rDNA were used to identify genetic affinity among 22 isolates of F. oxysporum f. sp. fagariae obtained from various location of major strawberry cultivating areas in Korea. Approximately 2.6kb DNA fragment was amplified with primer CNS1 and CNL12, and polymorphisms were observed with Avail and HinfI. A dendrogram was constructed using the UPGMA for cluster analysis. Eight distinct groups were clustered based on the banding pattern obtained from RAPD and rDNA RFLP. There was high level genetic variation among Korean isolates of Fusarium of sporum f. sp. fragariae.

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Phytophthora Root Rot of Chinese Cabbage and Spinach Caused by P. drechsleri in Korea

  • Jee, Hyeong-Jin;Kim, Wan-Gyn;Cho, Weon-Dae
    • The Plant Pathology Journal
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    • v.15 no.1
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    • pp.28-33
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    • 1999
  • Phytophthora root rot of Chniese cabbage and spinach is reported for the first time in Korea. The diseases ocurred at Yangju, Seosan and Yeocheon in Korea from 1995 through 1998, mainly in lowland and submerged areas. Symptoms consisted of stunt, yellows, wilt and eventual death due to root rot. Fourteen isolates collected from naturally infected plants were all identified as P. drechsleri based on mycological characteristics. PCR-RFLP analysis of rDNA of the isolates confirmed the above result, since the restriction band patterns of the small subunit and internal transcribed spacers were identical to P. drechsleri and P. cryptogea, but distinct from closely related species of P. erythroseptica, P. cambivora, P. sojae and P. megasperma. The pathogen showed strong pathogenicity to Chinese cabbage, moderate to spinach, radish, cabbage and tomato, and weak or none to brown mustard, kale, chicory and pepper in pathogenicity tests.

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Antifungal Activity in Cell-Free Culture Fluid of Pseudomons solanacearum Strains Collected from Severe Provinces in the North of Vietnam.

  • Cuong, Nguyen-Ngoc;Kieu, Le-Nhu;Hang, Dao thi-Thu;Long, Hoang-Hoa;Ha, Nguyen-Hong;Nhung, Vu-Thi;Minh, Le-Thi;Thanh
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.172-173
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    • 1998
  • A research collection of Pseudomons solanacearum bacteria, a pathogen causing ‘bacteria wilt’ disease of more than 265 plant species, represented for northern provinces of Vietnam has recently been established and was saved for examination of antifungal activity in their culture fluids. All strains used in this work have been isolated from infected tomato, potato, and groundnut collected from production fields and they express different levels of virulence on their host plants. Cell-free culture fluids of these strains were tested for antifungal activity (to inhibit growth of mycelium and to destroy germination tube of fungal spores) on a number of fungi that either infect or associate with vegetable crops of Solanaceae family (tomato, potato, pepers...), fruit plants (banana), and even well-known by Vietnamese traditional medicine herbal plants belonging to Trifoliatus, Schefflera, Homalomena and Panax genera (Araliaceae family) of which roots are used as a resource of the herbal material. The antifungal activity was found in nearly all strains tested. Result of study on chitin, CMC, tween 80 and casein degradation abilities of the latter suggested that antifungal activity of positively-found strains may be due to their ability of extracelluar chitinase's excretion that destroy fungal cell wall.

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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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Assessment of Virulence of Raffaelea quercus-mongolicae and Raffaelea spp. Isolates by Artificial Inoculation of Quercus mongolica Logs (Raffaelea quercus-mongolicae와 Raffaelea spp. 인공접종에 의한 신갈나무 줄기에서의 병원성 평가)

  • Son, Su-Yeon;Seo, Sang-Tae;Park, Hyung-Cheol
    • Research in Plant Disease
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    • v.20 no.4
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    • pp.270-274
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    • 2014
  • Division of Forest Diseases & Insect Pests, Korea Forest Research Institute, Seoul 130-712, Korea The objective of this study was to evaluate the virulence of geographically different isolates of oak wilt pathogen, Raffaelea quercus-mongolicae and other Raffaelea species. In this study, mature trees of Quercus mongolica were inoculated with the various isolates of Raffaelea spp. and their virulence was evaluated by measuring the extent of sapwood discoloration resulting from the inoculation. The average length of discolored sapwood in a lateral direction was longest in the trees inoculated with the isolates from Korea (8.69 cm) followed by R. quercivora (7.51 cm) and the other Raffaelea spp. (3.35 cm). The lateral length of discolored sapwood caused by the inoculation with Korean strains varied from 4.71 to 14.90 cm indicating their differences in virulence. The area of discolored sapwood caused by the inoculation with Raffaelea spp. varied from 1.57 to $8.42cm^2$ indicating their differences in virulence. Based on the length and area of the discolored sapwoods, isolated YY and wj43 appeared to have the highest virulence among all the Raffaelea isolates tested. Each of the two isolates was obtained from Gangwon Province and Jeonbuk Province, respectively.