• Title/Summary/Keyword: In vitro inoculation

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Expression and Promoter Analyses of Pepper CaCDPK4 (Capsicum annuum calcium dependent protein kinase 4) during Plant Defense Response to Incompatible Pathogen

  • Chung, Eun-Sook;Oh, Sang-Keun;Park, Jeong-Mee;Choi, Do-Il
    • The Plant Pathology Journal
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    • v.23 no.2
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    • pp.76-89
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    • 2007
  • CaCDPK4, a full-length cDNA clone encoding Capsicum annuum calcium-dependent protein kinase 4, was isolated from chili pepper (Capsicum annuum L.). Deduced amino acid sequence of CaCDPK4 shares the highest homology with tobacco NpCDPK8 and chickpea CaCDPK2 with 79% identity. Genomic blot analyses revealed that CaCDPK4 is present as a single copy in pepper genome, but it belongs to a multigene family. CaCDPK4 was highly induced when pepper plants were inoculated with an incompatible bacterial pathogen. Induced levels of CaCDPK4 transcripts were also detected in pepper leaves by the treatment of ethephon, an ethylene-inducing agent, and high-salt stress condition. The bacterial-expressed GST-CaCDPK4 protein showed to retain the autophosphorylation activity in vitro. GUS expression driven by CaCDPK4 promoter was examined in transgenic Arabidopsis containing transcriptional fusion of CaCDPK4 promoter. GUS expression under CaCDPK4 promoter was strong in the root and veins of the seedlings. GW (-1965) and D3 (-1377) promoters conferred on GUS expression in response to inoculation of an incompatible bacterial pathogen, but D4-GUS (-913) and DS-GUS (-833) did not. Taken together, our results suggest that CaCDPK4 can be implicated on signal transduction pathway of defense response against an incompatible bacterial pathogen in pepper.

Inhibitory Effect of IFN-$\beta$, on the Antitumor Activity of Celecoxib in U87 Glioma Model

  • Kim, Eun-Kyoung;Chung, Dong-Sup;Shin, Hye-Jin;Hong, Yong-Kil
    • Journal of Korean Neurosurgical Society
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    • v.46 no.6
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    • pp.552-557
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    • 2009
  • Objective : Interferon-$\beta$, (IFN-$\beta$) has been used in the treatment of cancers. Inhibition of the enzyme cyclooxygenase (COX) with celecoxib had a significantly suppressive effect on tumor growth, angiogenesis, and metastasis in a variety of tumors. The aim of this study was to elucidate the antiglioma effect of combined treatment with IFN-$\beta$ and celecoxib in U87 glioma model. Methods : The in vitro effects of IFN-$\beta$ (50-1,000 IU/mL) and celecoxib ($50-250\;{\mu}M$) alone or combination of both on the proliferation and apoptosis of U87 cells were tested using MTT assay, FACS analysis and DNA condensation. To determine the in vivo effect, nude mice bearing intracerebral U87 xenograft inoculation were treated with IFN-$\beta$ intraperitoneally ($2{\times}10^5\;IU/day$ for 15 days), celecoxib orally (5, 10 mg/kg) or their combination. Results : IFN-$\beta$ or celecoxib showed an inhibitory effect on the proliferation of U87 cells. When U87 cells were treated with IFN-$\beta$ and celecoxib combination, it seemed that IFN-$\beta$ interrupted the antiproliferative and apoptotic activity of celecoxib. No additive effect was observed on the survival of the tumor bearing mice by the combination of IFN-$\beta$ and celecoxib. Conclusion : These results suggest that IFN-$\beta$ seems to inhibit the antiglioma effect of celecoxib, therefore combination of IFN-$\beta$ and celecoxib may be undesirable in the treatment of glioma.

Spontaneous Morphological Transformation in Adenovirus Type 12 Induced Tumor Cells of Armenian and Chinese Hamsters (Adenovirus Type 12로 誘發한 Armenian Hamster 및 Chinese Hamster의 腫瘍細胞에서 보이는 形質轉換)

  • Kang, Yung-Sun;Sahsook Hahn
    • The Korean Journal of Zoology
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    • v.17 no.2
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    • pp.51-56
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    • 1974
  • Newborn Armenian and Chinese hamsters inoculated with adenovirus type 12 developed undifferentiated small cell tumors as early as 27 days after inoculation in the Chinese hamsters and within 30$\\sim$45 days in the Armenian hamsters. These tumors were transplantable and epithelial-like cell in morphology. Cultures of 6 tumors underwent spontaneous reversion to fibroblast-like morphology during the 14$\\sim$20 in vitro passages in the absence of chromosomal disturbances. While epithelial-like tumor derivatives were oncogenic and positive for the T-antigen, fibroblast-like revertants were non-oncogenic and negative for the T-antigen. Two other tumor derivatives reverted to fibroblast-like forms, immediately following exposure to SV40. These lacked the adenovirus T-antigen but were positive for the SV40 T-antigen and formed sarcomas in animals.

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Effects of sodium diacetate and microbial inoculants on fermentation of forage rye

  • Yan Fen Li;Eun Chan Jeong;Li Li Wang;Hak Jin Kim;Farhad Ahmadi;Jong Geun Kim
    • Journal of Animal Science and Technology
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    • v.65 no.1
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    • pp.96-112
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    • 2023
  • Rye (Secale cereale L.) is a valuable annual forage crop in Korea but there is limited information about the impact of chemical and biological additives on fermentation characteristics of the crop. This experiment was conducted to investigate fermentation dynamics of wilted forage rye treated with the following six additives; control (no additive), sodium diacetate applied at 3 g/kg wilted forage weight (SDA3), 6 g/kg wilted forage weight (SDA6), inoculations (106 CFU/g wilted forage) of Lactobacillus plantarum (LP), L. buchneri (LB), or LP+LB. The ensiled rye sampled at 1, 2, 3, 5, 10, 20, 30, and 45 days indicated that the acidification occurred fast within five days of storage than the rest of the storage period. The microbial inoculants decline the pH of ensiled forage, more rapidly than the control or SDA treated, which accompanied by the decrease of water-soluble carbohydrates and increase of lactic acid. Compared with the control silage, all treatments suppressed ammonia-nitrogen formation below to 35 g/kg DM throughout the sampling period. Suppression of total microbial counting occurred in SDA6, LP, and LP + LB. The lactic acid production rates were generally higher in microbial inoculation treatments. Acetic acid concentration was lowest in the LP-treated silage and highest in the SDA- and LB-treated silages. The in vitro dry matter (DM) digestibility and total digestible nutrients were the highest in the silage treated with SDA (6 g/kg) at day 45 of ensiling. Based on lower ammonia-nitrogen concentrations and higher feed value, ensiling forage rye treated with SDA at 6 g/kg is promising through enhanced silage quality.

Identification of Host-Resistant and Susceptible Varieties of Korean Grapes to Plasmopara viticola, a Pathogen Causing Grapevine Downy Mildew

  • Marc Semunyana;Sun Ha Kim;Jiyoung Min;Soo-Min Lee;Sang-Keun Oh
    • The Korean Journal of Mycology
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    • v.51 no.3
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    • pp.179-190
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    • 2023
  • Grapevine downy mildew, caused by Plasmopara viticola, significantly damages vineyards and is one of the most devastating diseases affecting cultivated grapes worldwide. In this study, we characterized the phenotypic and molecular traits of 11 P. viticola isolates from four grape-growing regions in South Korea. Additionally, we investigated the diversity of pathogenicity among these isolates and conducted an assay to evaluate the response of grape cultivars to P. viticola infection. Lemon-shaped sporangia were identified in the collected isolates, which released zoospores into the suspension at room temperature. Within a few hours of inoculation, the zoospores developed germ tubes. We tested 11 P. viticola isolates for pathogenicity in 845 grape cultivars to screen for grape host resistance to downy mildew infection. Among the tested isolates, JN-9 showed the highest virulence. Grape cultivars displayed varying phenotypic reactions to P. viticola infection: approximately 7% were highly susceptible, 41% were susceptible, 20% were moderately susceptible, 8% were resistant, and 24% exhibited extreme resistance. Phylogenetic analysis based on four genomic regions (internal transcribed spacer 1 [ITS1], actin, beta-tubulin, and cytochrome c oxidase II) revealed a close evolutionary relationship among all the Korean isolates, forming a single monophyletic lineage. Notably, these isolates showed greater similarity to European isolates than to American isolates. This comprehensive study contributes to a deeper understanding of the identity and behavior of P. viticola, which is crucial for developing effective resistance strategies against this pathogen in grape cultivars cultivated in South Korea.

Biological Control of Sclerotinia Rot Caused by Sclerotinia sclerotiorum in Lettuce Using Bacillus amyloliquefaciens HY (Bacillus amyloliquefaciens HY를 이용한 상추 균핵병의 생물학적 방제)

  • Mi-Ra Son;Jin-Won Kim
    • Research in Plant Disease
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    • v.30 no.3
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    • pp.247-255
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    • 2024
  • To test the antagonistic ability of Bacillus amyloliquefaciens HY against lettuce sclerotinia rot caused by Sclerotinia sclerotiorum, an in vitro test, greenhouse test and growth chamber test were conducted. As a result of in vitro dual culture test for sclerotia of B. amyloliquefaciens HY, the mycelial growth inhibition rate was 70.7%. Mycelial tip was swollen, black, and had an irregular and deformed shape. In I-plate test, the antagonistic volatile compounds produced by B. amyloliquefaciens HY, the mycelial growth inhibition rate was 57.7% and the sclerotia germination inhibition rate was 46.7%. In addition, B. amyloliquefaciens HY had sideropore production capacity, phosphate solubillibty, and protease production capability, but B. amyloliquefaciens HY did not produce chitinase. In growth chamber test, the lettuce survival rate was 0% in the control group treated with only the pathogen, and 80% in the treatment group treated with of S. sclerotiorum and B. amyloliquefaciens HY by diluting 10 times of the lettuce survival rate. In addition, the lettuce survival rate of 80% was shown in the treatment group treated with the culture medium of B. amyloliquefaciens HY 50 times 1 week later after inoculation with S. sclerotiorum.

Biological Management of Virulent Fusarium Species on Asparagus with Avirulent Fusarium Species In Vitro (비병원성(非病原性) Fusarium균(菌)을 이용(利用)한 아스파라거스의 병원성(病原性) Fusarium균(菌)의 생물적(生物的) 방제(防除))

  • Lee, Youn-Su
    • Korean Journal of Environmental Agriculture
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    • v.13 no.3
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    • pp.288-300
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    • 1994
  • Fusarium oxysporum was isolated most frequently, followed by F. moniliforme, and F. solani from infected asparagus plants grown in the field. In pathogenicity tests both with seedlings and plantlets, F. moniliforme showed higher virulence than Fusarium oxysporum did in general. Fusarium moniliforme showed more consistent virulence on both seedlings and plantlets than F. oxysporum did. Fusarium oxysporum showed higher virulence on plantlets than on seedlings. Fusarium solani showed very weak or no sign of virulence on seedlings and plantlets, respectively, in both tests. In protection tests with plantlets, most protection of asparagus against virulent fusarial infections occurred when challenge isolates were inoculated five or seven days after inoculation of protective fusarial species. Avirulent F. oxysporum was a more effective protective agent against infection of F. moniliforme than it was against F. oxysporum. Fusarium solani was more effective against infection of F. oxysporum than it was against F. moniliforme.

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Enhancement of Drought-Stress Tolerance of Brassica oleracea var. italica L. by Newly Isolated Variovorax sp. YNA59

  • Kim, Yu-Na;Khan, Muhammad Aaqil;Kang, Sang-Mo;Hamayun, Muhammad;Lee, In-Jung
    • Journal of Microbiology and Biotechnology
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    • v.30 no.10
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    • pp.1500-1509
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    • 2020
  • Drought is a major abiotic factor and has drastically reduced crop yield globally, thus damaging the agricultural industry. Drought stress decreases crop productivity by negatively affecting crop morphological, physiological, and biochemical factors. The use of drought tolerant bacteria improves agricultural productivity by counteracting the negative effects of drought stress on crops. In this study, we isolated bacteria from the rhizosphere of broccoli field located in Daehaw-myeon, Republic of Korea. Sixty bacterial isolates were screened for their growth-promoting capacity, in vitro abscisic acid (ABA), and sugar production activities. Among these, bacterial isolates YNA59 was selected based on their plant growth-promoting bacteria traits, ABA, and sugar production activities. Isolate YNA59 highly tolerated oxidative stress, including hydrogen peroxide (H2O2) and produces superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) activities in the culture broth. YNA59 treatment on broccoli significantly enhanced plant growth attributes, chlorophyll content, and moisture content under drought stress conditions. Under drought stress, the endogenous levels of ABA, jasmonic acid (JA), and salicylic acid (SA) increased; however, inoculation of YNA59 markedly reduced ABA (877 ± 22 ng/g) and JA (169.36 ± 20.74 ng/g) content, while it enhanced SA levels (176.55 ± 9.58 ng/g). Antioxidant analysis showed that the bacterial isolate YNA59 inoculated into broccoli plants contained significantly higher levels of SOD, CAT, and APX, with a decrease in GPX levels. The bacterial isolate YNA59 was therefore identified as Variovorax sp. YNA59. Our current findings suggest that newly isolated drought tolerant rhizospheric Variovorax sp. YNA59 is a useful stress-evading rhizobacterium that improved drought-stress tolerance of broccoli and could be used as a bio-fertilizer under drought conditions.

Laboratory passage and characterization of an isolate of Toxoplasma gondii from an ocular patient in Korea

  • Chai, Jong-Yil;Lin, Aifen;Shin, Eun-Hee;Oh, Myoung-Don;Han, Eun-Taek;Nam, Ho-Woo;Lee, Soon-Hyung
    • Parasites, Hosts and Diseases
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    • v.41 no.3
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    • pp.147-154
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    • 2003
  • Toxoplasma gondii tachyzoites were isolated from the blood of an ocular patient, and have been successfully passaged in the laboratory, for over a year, by peritoneal inoculation in mice. The isolated parasite was designated the Korean Isolate-1 (KI-1) and its characteristics were compared with those of the RH strain, a well-known virulent strain originating from a child who suffered from encephalitis. The morphology, pathogenicity, infectivity and cell culture characteristics of the KI-1 were similar to those of the RH strain. Both RH and KI-1 antigens were detected by an anti-T gondii monoclonal antibody (mAb), Tg563, against the major surface protein SAG1 (30 kDa), whereas no reaction was observed against an anti-Neospora caninum mAb, 12B4. The KI-1 was confirmed as an isolate of T. gondii. A long-term laboratory maintenance and characterization of a local T gondii isolate is reported for the first time in the Republic of Korea.

N-acetylcysteine and the human serum components that inhibit bacterial invasion of gingival epithelial cells prevent experimental periodontitis in mice

  • Alam, Jehan;Baek, Keum Jin;Choi, Yun Sik;Kim, Yong Cheol;Choi, Youngnim
    • Journal of Periodontal and Implant Science
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    • v.44 no.6
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    • pp.266-273
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    • 2014
  • Purpose: We previously reported that human serum significantly reduces the invasion of various oral bacterial species into gingival epithelial cells in vitro. The aims of the present study were to characterize the serum component(s) responsible for the inhibition of bacterial invasion of epithelial cells and to examine their effect on periodontitis induced in mice. Methods: Immortalized human gingival epithelial (HOK-16B) cells were infected with various 5- (and 6-) carboxy-fluorescein diacetate succinimidyl ester-labeled oral bacteria, including Fusobacterium nucleatum, Provetella intermedia, Porphyromonas gingivalis, and Treponiema denticola, in the absence or presence of three major serum components (human serum albumin [HSA], pooled human IgG [phIgG] and ${\alpha}1$-antitrypsin). Bacterial adhesion and invasion were determined by flow cytometry. The levels of intracellular reactive oxygen species (ROS) and activation of small GTPases were examined. Experimental periodontitis was induced by oral inoculation of P. gingivalis and T. denticola in Balb/c mice. Results: HSA and phIgG, but not ${\alpha}1$-antitrypsin, efficiently inhibited the invasion of various oral bacterial species into HOK-16B cells. HSA but not phIgG decreased the adhesion of F. nucleatum onto host cells and the levels of intracellular ROS in HOK-16B cells. N-acetyl-cysteine (NAC), a ROS scavenger, decreased both the levels of intracellular ROS and invasion of F. nucleatum into HOK-16B cells, confirming the role of ROS in bacterial invasion. Infection with F. nucleatum activated Rac1, a regulator of actin cytoskeleton dynamics. Not only HSA and NAC but also phIgG decreased the F. nucleatum-induced activation of Rac1. Furthermore, both HSA plus phIgG and NAC significantly reduced the alveolar bone loss in the experimental periodontitis induced by P. gingivalis and T. denticola in mice. Conclusions: NAC and the serum components HSA and phIgG, which inhibit bacterial invasion of oral epithelial cells in vitro, can successfully prevent experimental periodontitis.