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Characterization of the Maize Stalk Rot Pathogens Fusarium subglutinans and F. temperatum and the Effect of Fungicides on Their Mycelial Growth and Colony Formation

  • Shin, Jong-Hwan;Han, Joon-Hee;Lee, Ju Kyong;Kim, Kyoung Su
    • The Plant Pathology Journal
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    • v.30 no.4
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    • pp.397-406
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    • 2014
  • Maize is a socioeconomically important crop in many countries. Recently, a high incidence of stalk rot disease has been reported in several maize fields in Gangwon province. In this report, we show that maize stalk rot is associated with the fungal pathogens Fusarium subglutinans and F. temperatum. Since no fungicides are available to control these pathogens on maize plants, we selected six fungicides (tebuconazole, difenoconazole, fluquinconazole, azoxystrobin, prochloraz and kresoxim-methyl) and examined their effectiveness against the two pathogens. The in vitro antifungal effects of the six fungicides on mycelial growth and colony formation were investigated. Based on the inhibition of mycelial growth, the most toxic fungicide was tebuconazole with 50% effective concentrations ($EC_{50}$) of < $0.1{\mu}g/ml$ and $EC_{90}$ values of $0.9{\mu}g/ml$ for both pathogens, while the least toxic fungicide was azoxystrobin with $EC_{50}$ values of 0.7 and $0.5{\mu}g/ml$ for F. subglutinans and F. temperatum, respectively, and $EC_{90}$ values of > $3,000{\mu}g/ml$ for both pathogens. Based on the inhibition of colony formation by the two pathogens, kresoxim-methyl was the most toxic fungicide with complete inhibition of colony formation at concentrations of 0.1 and $0.01{\mu}g/ml$ for F. subglutinans and F. temperatum, respectively, whereas azoxystrobin was the least toxic fungicide with complete inhibition of colony formation at concentrations > $3,000{\mu}g/ml$ for both pathogens.

Characterization of dihydroflavonol 4-reductase cDNA in tea [Camellia sinensis (L.) O. Kuntze]

  • Singh, Kashmir;Kumar, Sanjay;Yadav, Sudesh Kumar;Ahuja, Paramvir Singh
    • Plant Biotechnology Reports
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    • v.3 no.1
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    • pp.95-101
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    • 2009
  • Tea leaves are major source of catechins—antioxidant flavonoids. Dihydroflavonol 4-reductase (DFR, EC 1.1.1.219) is one of the important enzymes that catalyzes the reduction of dihydroflavonols to leucoanthocyanins, a key ''late'' step in the biosynthesis of catechins. This manuscript reports characterization of DFR from tea (CsDFR) that comprised 1,413 bp full-length cDNA with ORF of 1,044 bp (115-1,158) and encoding a protein of 347 amino acids. Sequence comparison of CsDFR with earlier reported DFR sequences in a database indicated conservation of 69-87% among amino acid residues. In silico analysis revealed CsDFR to be a membrane-localized protein with a domain (between 16 and 218 amino acids) resembling the NAD-dependent epimerase/dehydratase family. The theoretical molecular weight and isoelectric point of the deduced amino sequence of CsDFR were 38.67 kDa and 6.22, respectively. Upon expression of CsDFR in E. coli, recombinant protein was found to be functional and showed specific activity of 42.85 nmol $min^{-1}$ mg $protein^{-1}$. Expression of CsDFR was maximum in younger rather than older leaves. Expression was down-regulated in response to drought stress and abscisic acid, unaffected by gibberellic acid treatment, but up-regulated in response to wounding, with concomitant modulation of catechins content. This is the first report of functionality of recombinant CsDFR and its expression in tea.

Stabilization of Metals-contaminated Farmland Soil using Limestone and Steel Refining Slag

  • Lim, Jeong-Muk;You, Youngnam;Kamala-Kannan, Seralathan;Oh, Sae-Gang;Oh, Byung-Taek
    • Journal of Soil and Groundwater Environment
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    • v.19 no.5
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    • pp.1-8
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    • 2014
  • The metals contamination of farmland soil nearby abandoned metal mine was serious problem in Korea. Stabilization of contaminated soil was reported using various stabilizers. Application of limestone and steel refining slag was reported as effective stabilizers in the stabilization of metals. The batch studies confirmed that the mixture of limestone and steel refining slag was suitable for stabilization of metals in contaminated soil. The limestone and steel refining slag mixture (2 : 1 and 3 : 2) were used in column studies and it was confirmed that the stabilizers effectively stabilized heavy metals in contaminated soil. The pH of the soil was increased with the addition of stabilizers. Total leached concentration of metals from the column study was reduced 44, 17, and 93% in comparison to the control at arsenic, cadmium and copper, respectively. The sequential extraction studies showed that the exchangeable fraction was changed into carbonate bound fraction (Cd and Cu) and Fe-Mn oxide bound fraction (As). Based on the results we confirmed that 2:1 ratio of limestone and steel refining slag effectively stabilizes the heavy metals. The mixed treatment of lime stone with steel refining slag would be an effective and feasible method for controlling metals leaching in contaminated soil.

Inhibitory Effect of Benzofuran Compound on Cyclooxygenase

  • Min, Kyung-Rak;Ahn, Ki-Young;Chung, Eun-Yong;Lee, Yong-Rok;Kim, Yeong-Shik;Kim, Young-Soo
    • Natural Product Sciences
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    • v.10 no.6
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    • pp.315-320
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    • 2004
  • Alpha-viniferin was previously isolated as a cyclooxygenase (COX)-2 inhibitor from Carex humilis (Cyperaceae) and is an oligomeric stilbene compound with benzofuran (BF) moieties in its chemical structure. In the present study, a chemically synthetic BF compound, named as 3,3-dimethyl-2,3,4,6,7,8,9,10,11,12,13,14,15,16,17,18-hexadecahydro-1H-benzo[b] cyclopentadeca[d]furan-1-one, was discovered to inhibit bacterial lipo polysaccharide (LPS)-induced prostaglandin $E_2$ $(PGE_2)$ production in macrophages RAW 264.7. The BF compound exhibited a selectively preferred inhibitory effect on COX-2 activity over COX-1 activity. Furthermore, BF compound inhibited LPS-induced COX-2 expression at transcription level. As a down-regulatory mechanism of COX-2 expression shown by BF compound, suppression of nuclear factor $(NF)-{\kappa}B$ activation has been demonstrated. BF compound inhibited LPS-induced $NF-{\kappa}B$ transcriptional activity and nuclear translocation of $NF-{\kappa}B$ p65, in parallel, but did not affect LPS-induced degradation of inhibitory ${\kappa}B{\alpha}$ protein $(I{\kappa}B{\alpha})$. Taken together, anti-inflammatory effect of BF compound on $PGE_2$ production was ascribed by its down-regulatory action on LPS-induced COX-2 synthesis in addition to inhibitory action on enzyme activity of COX-2.

Pseudomonas syringae pv. tomato DC3000 Improves Escherichia coli O157:H7 Survival in Tomato Plants

  • Namgung, Min;Lim, Yeon-Jeong;Kang, Min Kyu;Oh, Chang-Sik;Park, Duck Hwan
    • Journal of Microbiology and Biotechnology
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    • v.29 no.12
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    • pp.1975-1981
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    • 2019
  • Recently, outbreaks of food-borne diseases linked to fresh produce have been an emerging public health concern worldwide. Previous research has shown that when human pathogens co-exist with plant pathogens, they have improved growth and survival rates. In this study, we have assessed whether Escherichia coli O157:H7 benefits from the existence of a phytopathogenic bacterium and the underlying mechanisms were further investigated. When Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) and E. coli O157:H7 were co-inoculated by either dipping or infiltration methods, the populations of E. coli O157:H7 increased; however, no effect was observed when type three secretion system (T3SS) mutants were used instead, suggesting that E. coli O157:H7 benefits from the presence of Pst DC3000. In addition, this study confirmed that the E. coli O157:H7 populations increased when they occupied the tomato leaf intercellular space; this colonization of the interior of the leaves was possible due to the suppression of the PAMP-triggered immunity (PTI) by Pst DC3000, in particular with the AvrPto effector. In conclusion, our data support a plausible model that E. coli O157:H7 benefits from the presence of Pst DC3000 via AvrPto suppression of the PTI resistance.

A Real-Time PCR Assay for the Quantitative Detection of Ralstonia solanacearum in Horticultural Soil and Plant Tissues

  • Chen, Yun;Zhang, Wen-Zhi;Liu, Xin;Ma, Zhong-Hua;Li, Bo;Allen, Caitilyn;Guo, Jian-Hua
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.193-201
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    • 2010
  • A specific and rapid real-time PCR assay for detecting Ralstonia solanacearum in horticultural soil and plant tissues was developed in this study. The specific primers RSF/RSR were designed based on the upstream region of the UDP-3-O-acyl-GlcNAc deacetylase gene from R. solanacearum, and a PCR product of 159 bp was amplified specifically from 28 strains of R. solanacearum, which represent all genetically diverse AluI types and all 6 biovars, but not from any other nontarget species. The detection limit of $10^2\;CFU/g$ tomato stem and horticultural soil was achieved in this real-time PCR assay. The high sensitivity and specificity observed with field samples as well as with artificially infected samples suggested that this method might be a useful tool for detection and quantification of R. solanacearum in precise forecast and diagnosis.

Effects of Adding Urea and Molasses on Napiergrass Silage Quality

  • Yunus, M.;Ohba, N.;Shimojo, M.;Furuse, M.;Masuda, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.11
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    • pp.1542-1547
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    • 2000
  • To standardize proper formulation of urea and molasses, the former to increase crude protein content of tropical grass and the latter for improving its silage quality, we examined the fermentation quality of silage of fresh and wilted napiergrass (Pennisetum purpureum Schumach) with different levels of urea and molasses with or without lactic acid bacteria (LAB). Silage was made of napiergrass with conditions of fresh young (Exp. 1),young wilted for half day (Exp. 2) and fresh mature (Exp. 3). Chopped plant materials of about 1cm length were ensiled into a laboratory silo and incubated for one month at $25^{\circ}C$. The treatments were the combination of 0, 0.2 and 0.6% of urea and 0, 2 and 5% of molasses (fresh material basis) with or without LAB inoculation. After opening the silo, pH, organic acids, volatile basic nitrogen (VBN) and total nitrogen (TN) were determined. Addition of molasses significantly (p<0.01) lowered pH values in three experiments. Though molasses addition increased lactic acid production even at a higher level of urea, pH values at 0 and 2% molasses were significantly increased by urea in fresh and wilted young silages, but in fresh mature silage it occurred only when molasses was not added. VBN/TN at 0.6% urea were decreased significantly by the highest molasses in three experiments. Significant increases in TN by the increasing of urea addition were observed at all levels of molasses in wilted young and fresh mature silages. In conclusion, a combination of 5% molasses and 0.6% urea could improve the nutritive and fermentation qualities of napiergrass silage under young, wilting and mature conditions.

Meta-analysis Reveals That the Genus Pseudomonas Can Be a Better Choice of Biological Control Agent against Bacterial Wilt Disease Caused by Ralstonia solanacearum

  • Chandrasekaran, Murugesan;Subramanian, Dharaneedharan;Yoon, Ee;Kwon, Taehoon;Chun, Se-Chul
    • The Plant Pathology Journal
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    • v.32 no.3
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    • pp.216-227
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    • 2016
  • Biological control agents (BCAs) from different microbial taxa are increasingly used to control bacterial wilt caused by Ralstonia solanacearum. However, a quantitative research synthesis has not been conducted on the role of BCAs in disease suppression. Therefore, the present study aimed to meta-analyze the impacts of BCAs on both Ralstonia wilt disease suppression and plant (host) growth promotion. The analysis showed that the extent of disease suppression by BCAs varied widely among studies, with effect size (log response ratio) ranging from -2.84 to 2.13. The disease incidence and severity were significantly decreased on average by 53.7% and 49.3%, respectively. BCAs inoculation also significantly increased fresh and dry weight by 34.4% and 36.1%, respectively on average. Also, BCAs inoculation significantly increased plant yield by 66%. Mean effect sizes for genus Pseudomonas sp. as BCAs were higher than for genus Bacillus spp. Among antagonists tested, P. fluorescens, P. putida, B. cereus, B. subtilis and B. amyloliquefaciens were found to be more effective in general for disease reduction. Across studies, highest disease control was found for P. fluorescens, annual plants, co-inoculation with more than one BCA, soil drench and greenhouse condition were found to be essential in understanding plant responses to R. solanacearum. Our results suggest that more efforts should be devoted to harnessing the potential beneficial effects of these antagonists, not just for plant growth promoting traits but also in mode of applications, BCAs formulations and their field studies should be considered in the future for R. solanacearum wilt disease suppression.

Anti-Inflammatory Mode of Isoflavone Glycoside Sophoricoside by Inhibition of Interleukin-6 and Cyclooxygenase-2 in Inflammatory Response

  • Kim, Byung-Hak;Chung, Eun-Yong;Ryu, Jae-Chun;Jung, Sang-Hun;Min, Kyung-Rak;Kim, Youngsoo
    • Archives of Pharmacal Research
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    • v.26 no.4
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    • pp.306-311
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    • 2003
  • Soy, high dietary intake for the oriental population, is a main source of isoflavonoids. Sophoricoside (SOP) an isoflavone glycoside was isolated from immature fruits of Sophora japonica (Leguminosae family) and its inhibitory effect on chemical mediators involved in inflammatory response was investigated in this study. SOP inhibited the interleukin (IL)-6 bioactivity with an $IC_{50}$ value of 6.1 $\mu$M whereas it had no effects on IL-1$\beta$ and TNF-a bioactivities. SOP was identified as a selective inhibitor of cyclooxygenase (COX)-2 activity with an $IC_{50}$ value of 4.4 $\mu$ M, but did not show inhibitory effect on the synthesis of COX-2. However, SOP had no effect on the production of reactive oxygen species including superoxide anions and nitric oxide. These results revealed that in vitro anti-inflammatory action of SOP is significantly different from that of genistein known as a phytoestrogen of soy products. This experimental study has documented an importance of dietary soy isoflavonoids as multifunctional agents beneficial to human health, and will help to clarify protective mechanisms of SOP against inflammatory conditions.

Effects of Preheated Additives on the Fermentation Quality of Napiergrass Silage

  • Yunus, M.;Ohba, N.;Tobisa, M.;Shimojo, M.;Masuda, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.11
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    • pp.1564-1567
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    • 2001
  • Hydrolysis of plant protein to non-protein nitrogen (N) or ammonia can reduce quality of silage crops. Heating or non-enzymatic browning is a treatment to inhibit this hydrolysis. This experiment was conducted to examine the effects of pre-heated soybean meal and molasses on the fermentation quality of napiergrass silage. The initial growth of napiergrass was harvested at 85 days of age and immediately chopped into about 1 cm length. About 700 g of the grass was ensiled into a laboratory silo (1.0 liter polyethylene container) and incubated for 30 days at room temperature ($28^{\circ}C$). No additives (control), molasses, soybean meal and molasses + soybean meal treatments were prepared. All additives were non-heated or heated in an oven at $150^{\circ}C$ for 30 minutes before ensiling. Molasses was added at 3% on the fresh weight basis and soybean meal was added at 0.5% N, respectively. After opening the silo, pH, total nitrogen (TN), volatile basic nitrogen (VBN), lactic acid (LA), acetic acid (AA), butyric acid (BA) and dry matter (DM) contents were determined. The data were analyzed statistically by analysis of variance. Compared with control, molasses addition significantly decreased pH value, VBN/TN, AA and BA and increased LA production. Soybean meal addition significantly increased TN and VBN/TN of silage. Both molasses and soybean meal addition significantly reduced pH value, AA, and BA and increased DM and LA contents of silage. The heating of additives was only effective to reduce VBN/TN production compared with non-heated additives in soybean meal and soybean meal with molasses addition.