• Title/Summary/Keyword: Biological Assay

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Evaluation of Rhizobacterial Isolates for Their Antagonistic Effects against Various Phytopathogenic Fungi (식물 근권에서 분리한 미생물의 식물병원성 진균에 대한 길항효과 검정)

  • Kim, Yun Seok;Kim, Sang woo;Lamsal, Kabir;Lee, Youn Su
    • The Korean Journal of Mycology
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    • v.44 no.1
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    • pp.36-47
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    • 2016
  • This study was conducted to evaluate five different strains of rhizobacterial isolates viz. PA1, PA2, PA4, PA5 and PA12 for biological control against Colletotrichum acutatum, C. coccodes, C. gloeosporioides, C. dematium, Botrytis cinerea, Rhizoctonia solani, Sclerotinia minor and Fusarium sp. In vitro inhibition assay was performed on three different growth mediums, potato dextrose agar (PDA), tryptic soy agar (TSA), and PDA-TSA (1:1 v/v) for the selection of potential antagonistic isolates. According to the result, isolate PA2 showed the highest inhibitory effect with 65.5% against C. coccodes on PDA and with 96.5% against S. minor on TSA. However, the same isolate showed the highest inhibition with 58.5% against C. acutatum on PDA-TSA. In addition, an in vivo experiment was performed to evaluate these bacterial isolates for biological control against fungal pathogens. Plants treated with bacteria were analyzed with phytopathogens and plants inoculated with phytopathogens were treated with isolates to determine the biological control effect against fungi. According to the result, all five isolates tested showed inhibitory effects against phytopathogens at various levels. Mode of action of these rhizobacterial isolates was evaluated with siderophore production, protease assay, chitinase assay and phosphate solubilizing assay. Bacterial isolates were identified by 16S rDNA sequencing, which showed that isolates PA1 and PA2 belong to Bacillus subtilis, whereas, PA4, PA5, and PA12 were identified as Bacilus altitudinis, Paenibacillus polymyxa and Bacillus amyloliquefaciens, respectively. Results of the current study suggest that rhizobacterial isolates can be used for the plant growth promoting rhizobacteria (PGPR) effect as well as for biological control of various phytopathogens.

Determination of Carbon Source Utilization of Bacillus and Pythium Species by Biolog$^{(R)}$ Microplate Assay

  • Chun, Se-Chul;R.W. Schneider;Chung, Ill-Min
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.252-258
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    • 2003
  • The carbon utilizations of Bacillus species and Pythium species were investigated by using a Biolog$^{(R)}$ microplate assay to determine if there are differences in the carbon utilizations of selected strains of these species. It may be possible to afford a competitive advantage to bacterial biological control agents by providing them with a substrate that they can readily use as a carbon source, for example, in a seed coating formulation. Microplates, identified as SFP, SFN and YT were used to identify spore-forming bacteria, nonspore-forming bacteria, and yeast, respectively. Bacterial and mycelial suspensions were adjusted to turbidities of 0.10 to 0.11 at 600 nm. One hundred microliters of each of the bacterial and mycelial suspension were inoculated into each well of each of the three types of microplates. L-arabinose, D-galactose, D-melezitose and D-melibiose of the 147 carbohydrates tested were found to be utilized only by bacteria, and not by Pythium species, by Biolog$^{(R)}$ microplate assay, and this was confirmed by traditional shake flask culture. Thus, it indicated that the Biolog$^{(R)}$ microplate assay could be readily used to search for specific carbon sources that could be utilized to increase the abilities of bacterial biological control agents to adapt to contrived environments.

In vitro Biological Activity Assay of Ethanol Extract of Radish (무 에탄올 추출물의 in vitro 생리활성 분석)

  • Jung, Min-Suk;Lee, Gun-Soon;Chae, Hee-Jeong
    • Applied Biological Chemistry
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    • v.47 no.1
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    • pp.67-71
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    • 2004
  • In vitro biological activities of ethanol extract of radish including whitening, hangover removal, antimicrobial and antioxidant activities were analyzed. For whitening activity assay, tyrosinase inhibition rate was measured as $IC_{50}$ (50% inhibitory concentration). The $IC_{50}$ values of radish trunk and root extracts were estimated as 0,9 mg/ml and 2.1 mg/ml, respectively. Radish trunk extract showed 2.5-fold tyrosinase inhibition activity of radish root extract, however, there was no significant difference according to radish species. By alcohol dehydrogenase (ADH) activity assay as a hangover removal activity assay, radish trunk extract (2.5 mg/ml) and root extracts (8 mg/ml) showed ]50% activation of ADH. TBA values of radish trunk and root extracts (1% of each) were 43-61 % level of ${\alpha}-tochoperol$ (2.2%). From the analysis of in vitro biological activities of radish, it was suggested that radish could be used in functional food or cosmetics containing hangover removal, whitening and antioxidant activities.

Using Chemical and Biological Approaches to Predict Energy Values of Selected Forages Affected by Variety and Maturity Stage: Comparison of Three Approaches

  • Yu, P.;Christensen, D.A.;McKinnon, J.J.;Soita, H.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.2
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    • pp.228-236
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    • 2004
  • Two varieties of alfalfa (Medicago sativa L cv. Pioneer and Beaver) and timothy (Phleum pratense L cv. Climax and Joliette), grown at different locations in Saskatchewan (Canada), were cut at three stages [1=one week before commercial cut (early bud for alfalfa; joint for timothy); 2=at commercial cut (late bud for alfalfa; pre-bloom head for timothy); 3=one week after commercial cut (early bloom for alfalfa; full head for timothy)]. The energy values of forages were determined using three approaches, including chemical (NRC 2001 formula) and biological approaches (standard in vitro and in situ assay). The objectives of this study were to determine the effects of forage variety and stage of maturity on energy values under the climate conditions of western Canada, and to investigate relationship between chemical (NRC 2001 formula) approach and biological approaches (in vitro and in situ assay) on prediction of energy values. The results showed that, in general, forage species (alfalfa vs. timothy) and cutting stage had profound impacts, but the varieties within each species (Pioneer vs. Beaver in alfalfa; Climax vs. Joliette in timothy) had minimal effects on energy values. As forage maturity increased, the energy contents behaved in a quadratic fashion, increasing at stage 2 and then significantly decreasing at stage 3. However, the prediction methods-chemical approach (NRC 2001 formula) and biological approaches (in vitro and in situ assay) had great influences on energy values. The highest predicted energy values were found by using the in situ approach, the lowest prediction value by using the NRC 2001 formula, and the intermediate values by the in vitro approach. The in situ results may be most accurate because it is closest to simulate animal condition. The energy values measured by biological approaches are not predictable by the chemical approach in this study, indicating that a refinement is needed in accurately predicting energy values.

Isolation, Identification and Biological Control Activity of SKU-78 Strain against Ralstonia solanacearum (풋마름병균, Ralstonia solanacearum의 길항세균 SKU-78 균주의 분리 동정 및 특성)

  • Sung, Pil-Je;Shin, Jeong-Kun;Cho, Hong-Bum;Kim, Shin-Duk
    • Applied Biological Chemistry
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    • v.48 no.1
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    • pp.48-52
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    • 2005
  • Six stains of plant growth promoting rhizobacteria were selected through germinating seed assay and root colonization assay. Among them, SKU-78 strain induced significant suppression of bacterial wilt disease in tomato and pepper plants. Seed treatment followed by soil drench application with this strain resulted in over 60% reduction of bacterial wilt disease compared with the control. It was suggested that SKU-78 strain activated the host defense systems in plants, based on lack of direct antibiosis against pathogen. According to Bergey's Manual of Systemic Bacteriology and 16S rDNA sequence data, SKU-78 stain was identified as Bacillus sp. SKU-78.

The N-terminal peptide of the main protease of SARS-CoV-2, targeting dimer interface, inhibits its proteolytic activity

  • Sunyu Song;Yeseul Kim;Kiwoong Kwak;Hyeonmin Lee;Hyunjae Park;Young Bong Kim;Hee-Jung Lee;Lin-Woo Kang
    • BMB Reports
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    • v.56 no.11
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    • pp.606-611
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    • 2023
  • The main protease (Mpro) of SARS-CoV-2 cleaves 11 sites of viral polypeptide chains and generates essential non-structural proteins for viral replication. Mpro is an important drug target against COVID-19. In this study, we developed a real-time fluorometric turn-on assay system to evaluate Mpro proteolytic activity for a substrate peptide between NSP4 and NSP5. It produced reproducible and reliable results suitable for HTS inhibitor assays. Thus far, most inhibitors against Mpro target the active site for substrate binding. Mpro exists as a dimer, which is essential for its activity. We investigated the potential of the Mpro dimer interface to act as a drug target. The dimer interface is formed of domain II and domain III of each protomer, in which N-terminal ten amino acids of the domain I are bound in the middle as a sandwich. The N-terminal part provides approximately 39% of the dimer interface between two protomers. In the real-time fluorometric turn-on assay system, peptides of the N-terminal ten amino acids, N10, can inhibit the Mpro activity. The dimer interface could be a prospective drug target against Mpro. The N-terminal sequence can help develop a potential inhibitor.

Nanobiocatalyst-Linked Immunosorbent Assay(NBC-LISA) (나노바이오촉매 기반 효소결합면역흡착검사)

  • Lee, Inseon;Hwang, Sang Youn;Kim, Jungbae
    • Korean Chemical Engineering Research
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    • v.49 no.4
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    • pp.387-392
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    • 2011
  • Enzymes are being used in various fields due to their unique property of substrate specificity. Enzyme-linked immunosorbent assay(ELISA) has enabled the detection of various antigens by reporting the binding event of antigen and antibody via enzyme-catalyzed reaction. However, the sensitivity improvement of conventional ELISA has been limited because only one enzyme molecule is conjugated to one molecule of antibody. To overcome this limitation and further improve the sensitivity of ELISA, there have been efforts to increase the number ratio of enzymes to antibody. Recently, the nanobiocatalytic approaches, with their successful enzyme stabilization, improved the performance stability as well as sensitivity in a modified protocol of ELISA. The present paper introduces the basic principle of ELISA, and the recent efforts to improve sensitivity and performance stability of ELISA by using the nanobiocatalytic approaches.