• Title/Summary/Keyword: Bioassays

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Experimental Analysis of Interactions Among Saprotrophic Fungi from A Phosphorous-Poor Desert Oasis in the Chihuahuan Desert

  • Marini-Macouzet, Constanza;Munoz, Luis;Gonzalez-Rubio, Aldo;Eguiarte, Luis E.;Souza, Valeria;Velez, Patricia
    • Mycobiology
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    • v.48 no.5
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    • pp.410-417
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    • 2020
  • Fungal ecological interactions play a key role in structuring community assemblages. These associations may involve both antagonistic and synergistic relationships, which are commonly influenced by abiotic factors such as nutrient conditions. However, information for extreme, oligotrophic systems remain poor. Herein, interactions among key members of the aquatic transient fungal community (Aspergillus niger, Cladosporium sp., and Coprinellus micaceus) of a low-nutrient freshwater system in the Cuatro Ci enegas Basin, Mexico were studied. Pairwise interaction bioassays were explored in vitro under different nutrient conditions, including carbohydrates-rich, carbohydrates and amino peptides-rich, and low nutrients. Our results indicated that antagonistic patterns prevail among the studied taxa. However, nutrient-dependent changes were observed in Cladosporium sp. shifting to synergy under carbohydrates-rich conditions, suggesting changes in the fungal community composition as a result of nutrient enrichment. Remarkably, our findings contrast with previous work demonstrating mainly synergistic interactions between our tested fungal isolates and co-occurring autochthonous bacteria (Aeromonas spp. and Vibrio sp.) under low-nutrient conditions. This observation may indicate that bacteria and fungi exhibit distinct community-level responses, driven by nutrient conditions. This contributes to the knowledge of fungal community dynamics and interspecific interactions in an oligotrophic ecosystem, highlighting the relevance of nutrient-based shifts and antagonistic interactions in ecosystem dynamics.

Synergistic Phosphate Solubilization by Burkholderia anthina and Aspergillus awamori

  • Walpola, Buddhi Charana;Jang, Hyo-Ju;Yoon, Min-Ho
    • Korean Journal of Soil Science and Fertilizer
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    • v.46 no.2
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    • pp.117-121
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    • 2013
  • Single or co-inoculation of phosphate solubilizing bacterial and fungal strains (Burkholderia anthina and Aspergillus awamori respectively) was performed separately to assess their synergistic and antagonistic interactions and the potential to be used as bio-inoculants. Co-inoculation was found to release the highest content of soluble phosphorus (1253 ${\mu}g\;ml^{-1}$) into the medium, followed by single inoculation of fungal strain (1214 ${\mu}g\;ml^{-1}$) and bacterial strain (997 ${\mu}g\;ml^{-1}$). However, there was no significant difference between single inoculation of fungal strain and co-inoculation of fungal and bacterial strain in terms of the phosphorous release. The highest pH reduction, organic acid production and glucose consumption were observed in the sole A. awamori inoculated culture medium. According to the plant growth promotion bioassays, co-inoculation of the microbial strains resulted in 21% and 43% higher shoot and root growth of the mung bean seedlings respectively as compared to the respective controls. Therefore, co-inoculation of B. anthina and A. awamori showed better performance in stimulating plant growth than that in inoculation of each strain alone. However, assessment period of the present study being short, we recommend in engaging further experimentation under field conditions in order to test the suitability of the strains to be used as bio-inoculants.

Investigations on Bacteria as a Potential Biological Control Agent of Summer Chafer, Amphimallon solstitiale L. (Coleoptera: Scarabaeidae)

  • Sezen Kazlm;Demir Ismail;Katl Hatice;Demirbag Zihni
    • Journal of Microbiology
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    • v.43 no.5
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    • pp.463-468
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    • 2005
  • Studying the bacteria of hazardous insects allows the opportunity to find potentially better biological control agents. Therefore, in this study, bacteria from summer chafer (Amphimallon solstitiale L., Coleoptera: Scarabaeidae) we isolated and identified the insecticidal effects of bacteria isolated from A. solstitiale and Melolontha melolontha L. (common cockchafer, Coleoptera: Scarabaeidae) and the mixtures of these bacterial isolates were investigated on A. solstitiale larvae. Crystals from Bacillus sp. isolated from M. melolontha were also purified, and tested against the second and third-stage larvae of A. solstitiale. The bacterial isolates of A. solstitiale were identified as Pseudomonas sp., Pseudomonas sp., Bacillus cereus and Micrococcus luteus, based on their morphology, spore formation, nutritional features, and physiological and biochemical characteristics. The insecticidal effects of the bacterial isolates determined on the larvae of A. solstitiale were $90\%$ with B. cereus isolated from A. solstitiale, and $75\%$ with B. cereus, B. sphaericus and B. thuringiensis isolated from M. melolontha within ten days. The highest insecticidal effects of the mixed infections on the larvae of A. solstitiale were $100\%$ both with B. cereus+B. sphaericus and with B. cereus+B. thuringiensis. In the crystal protein bioassays, the highest insecticidal effect was $65\%$ with crystals of B. thuringiensis and B. sphaericus isolated from M. melolontha within seven days. Finally, our results showed that the mixed infections could be utilized as microbial control agents, as they have a $100\%$ insecticidal effect on the larvae of A. solstitiale.

Antagonistic Activity of Bacteria Isolated from Apple in Different Fruit Development Stages against Blue Mold Caused by Penicillium expansum

  • Lopez-Gonzalez, Rocio Crystabel;Juarez-Campusano, Yara Suhan;Rodriguez-Chavez, Jose Luis;Delgado-Lamas, Guillermo;Medrano, Sofia Maria Arvizu;Martinez-Peniche, Ramon Alvar;Pacheco-Aguilar, Juan Ramiro
    • The Plant Pathology Journal
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    • v.37 no.1
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    • pp.24-35
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    • 2021
  • Blue mold caused by Penicillium expansum is one of the most significant postharvest diseases of apples. Some microorganisms associated with the surface of ripening apples possess the ability to inhibit the growth of P. expansum. However, the existing literature about their colonization in the stages before ripening is not explored in depth. This study aims to characterize the antagonistic capacity of bacterial populations from five fruit development stages of 'Royal Gala' apples. The results have shown that the density of the bacterial populations decreases throughout the ripening stages of fruit (from 1.0 × 105 to 1.1 × 101 cfu/㎠). A total of 25 bacterial morphotypes (corresponding to five genera identified by 16S RNA) were differentiated in which Bacillus stood out as a predominant genus. In the in vitro antagonism tests, 10 Bacillus strains (40%) inhibited the mycelial growth of P. expansum from 30.1% to 60.1%, while in fruit bioassays, the same strains reduced the fruit rot ranging from 12% to 66%. Moreover, the bacterial strains with antagonistic activity increased in the ripening fruit stage. B. subtilis subsp. spiziennii M24 obtained the highest antagonistic activity (66.9% of rot reduction). The matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis revealed that bacteria with antagonistic activity produce anti-fungal lipopeptides from iturin and fengycin families.

The Study on the Physiological Activities of Beta vulgaris such as Antioxidant and Anti-inflammatory in RAW 264.7 cells (RAW 264.7 세포에서 레드비트의 항산화 및 항염증 등의 생리활성 연구)

  • Ji, Joong-Gu
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.1
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    • pp.309-317
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    • 2021
  • This study was designed to examine the cell cytotoxic, anti-inflammatory, and antioxidant activity for raw material of Beta vulagaris tea. Antioxidative ability was evaluated by bioassays using DPPH and ABTS radical scavenging activity. Cell viability was assessed by MTT assay using RAW 264.7 cells, and investigated production levels of reactive oxide speies, and inflammatory meditors(i.e., nitric oxide, tumor necrosis factor-α, and interleukin-6) in LPS-induced RAW 264.7 cells. As a results, DPPH and ABTS raidcal scavenging activity were increased in a dose-dependent manner, and confirmed no cytotoxicity in all concentration. Also, it was significantly decreased level of ROS, NO, IL-6, and TNF-a in LPS-induced RAW 264.7 cells. Therefore these results suggest Beta vulagaris has considerable potential as a raw material of leached tea with safe anti-oxidative and anti-inflammatory effects.

Anti-Glyaction and Radical Scavenging Activities of Ligustri Fructus by Extraction Method (추출 방법에 따른 여정실의 최종당화산물 생성 저해 및 라디칼 소거 활성)

  • Jeong, Yun Hee;Kim, Seo Yoon;Jeong, Gyeong Han;Kim, Tae Hoon
    • The Korea Journal of Herbology
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    • v.33 no.4
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    • pp.95-100
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    • 2018
  • Objectives : Ligustri Fructus has been used since ancient times as a medicinal usages in folk medicines against antitumor purpose. Many biological active constituents have been identified from this biomass such as several terpenoids and lignans. In current study, the properties of antioxidant and anti-diabetic complications using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)($ABTS^+$) radicals scavenging, and advanced glycation end products (AGEs) inhibition assays were evaluated by different extraction methods of Ligustri Fructus. Methods : In present continuing research for development of bioactive natural ingredients, antioxidant and AGEs formation inhibitory capacities of Ligustri Fructus extracts using different organic solvents were prepared and the biological potentials were investigated using in vitro bioassays. Antioxidant properties were evaluated employing radical scavenging assays using DPPH and $ABTS^+$ radicals. In addition, the anti-diabetic complications effects of Ligustri Fructus extracts were tested via AGEs formation inhibitory assay. The total phenolic contents were determined using a spectrophotometric method. Results : All the tested extracts exhibited dose-dependent radical scavenging and AGEs formation inhibitory activities. Among the tested samples, hot water extract of Ligustri Fructus was showed the most potent activity with $IC_{50}$ value of $494.8{\pm}6.7{\mu}g/m{\ell}$ against DPPH radical scavenging assay. Also, $ABTS^+$ radical scavenging activity of hot water extract was higher than those of other extracts. In addition, AGEs formation inhibitory effects of each extacts and total phenolic contents were evaluated. Conclusions : These results suggested that Ligustri Fructus can be considered as a new effective source of natural antioxidant and anti-diabetic complications resources.

Effects of Pahs and Pcbs and Their Toxic Metabolites on Inhibition of Gjic and Cell Proliferation in Rat Liver Epithelial Wb-F344 Cells

  • Miroslav, Machala;Jan, Vondracek;Katerina, Chramostova;Lenka, Sindlerova;Pavel, Krcmar;Martina, Pliskova;Katerina, Pencikova;Brad, Upham
    • Environmental Mutagens and Carcinogens
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    • v.23 no.2
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    • pp.56-62
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    • 2003
  • The liver progenitor cells could form a potential target cell population fore both tumor-initiating and -promoting chemicals. Induction of drug-metabolizing and antioxidant enzymes, including AhR-dependent CYP1A1, NQO-1 and AKR1C9, was detected in the rat liver epithelial WB-F344 "stem-like" cells. Additionally, WB-F344 cells express a functional, wild-type form of p53 protein, a biomarker of genotoxic events, and connexin 43, a basic structural unit of gap junctions forming an important type of intercellular communication. In this cellular model, two complementary assays have been established for detection of the modes of action associated with tumor promotion: inhibition of gap junctional intercellular communication (GJIC) and proliferative activity in confluent cells. We found that the PAHs and PCBs, which are AhR agonists, released WB-F344 cells from contact inhibition, increasing both DNA synthesis and cell numbers. Genotoxic effects of some PAHs that lead to apoptosis and cell cycle delay might interfere with the proliferative activity of PAHs. Contrary to that, the nongenotoxic low-molecular-weight PAHs and non-dioxin-like PCB congeners, abundant in the environment, did not significantly affect cell cycle and cell proliferation; however both groups of compounds inhibited GJIC in WB-F344 cells. The release from contact inhibiton by a mechanism that possibly involves the AhR activation, inhibition of GJIC and genotoxic events induced by environmental contaminants are three important modes of action that could play an important role in carcinogenic effects of toxic compounds. The relative potencies to inhibit GJIC, to induce AhR-mediated activity, and to release cells from contact inhibition were determined for a large series of PAHs and PCBs and their metabolites. In vitro bioassays based on detection of events on cellular level (deregulation of GJIC and/or proliferation) or determination of receptor-mediated activities in both ?$stem-like^{\circ}{\times}$ and hepatocyte-like liver cellular models are valuable tools for detection of modes of action of polyaromatic hydrocarbons. They may serve, together with concentration data, as a first step in their risk assessment.

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Interaction between Brassinolide and Auxins on Bioassays (Brassinolide와 Auxin과의 상호작용의 생물검정에 의한 평가)

  • Choi, Chung-Don;Kim, Soon-Chul;Lee, Soo-Kwan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.35 no.1
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    • pp.58-64
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    • 1990
  • This study was conducted to investigate the interaction effect of a new plant growth regulator brassinolide and auxins using several bioassay methods such as rice lamina inclination test, oat coleoptile segment growth test and radish hypocotyl elongation test. For rice lamina inclination test, the antagonistic response showed at low concentration combinations of homobrassinolide and auxins (IAA and 2,4-D), while the combinations of high concentration responsed mostly synergistic or additive effects, IAA was generally higher than 2,4-D for combination effect with homobrassinolide. For oat coleoptile segment growth test, the synergistic effect showed at IAA concentrations less than 0.1ppm while additive response exhibited above 0.3ppm regardless of homobrassinolide concentrations. In radish hypocotyl elongation test, the interaction response varied with cotyledon. The sections removed cotyledon showed mostly antagonistic effects, except for combination of homobrassinolide with IAA 3ppm. Interaction responses were Quite different from bioassay methods, particularly using experimental materials: antagonistic responses were usually shown at the section that attached growing point while these for sections that removed growing point were responded synergistic or additive effect due to unstable endogenous hormones.

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Identification and Characterization of Pseudomonas syringae pv. syringae, a Causative Bacterium of Apple Canker in Korea

  • Seunghee, Lee;Wonsu, Cheon;Hyeok Tae, Kwon;Younmi, Lee;Jungyeon, Kim;Kotnala, Balaraju;Yongho, Jeon
    • The Plant Pathology Journal
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    • v.39 no.1
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    • pp.88-107
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    • 2023
  • In the present investigation, bacterial isolates from infected apple trees causing apple canker during winter were studied in the northern Gyeongbuk Province, Korea. The pathogen was identified as Pseudomonas syringae pv. syringae (Pss) through various physiological and biochemical characterization assays such as BIOLOG, gas chromatography of fatty acid methyl esters, and 16S rRNA. Bioassays for the production of phytotoxins were positive for syringopeptin and syringomycin against Bacillus megaterium and Geotrichum candidum, respectively. The polymerase chain reaction (PCR) method enabled the detection of toxin-producing genes, syrB1, and sypB in Pss. The differentiation of strains was performed using LOPAT and GATTa tests. Pss further exhibited ice nucleation activity (INA) at a temperature of -0.7℃, indicating an INA+ bacterium. The ice-nucleating temperature was -4.7℃ for a non-treated control (sterilized distilled water), whereas it was -9.6℃ for an INA- bacterium Escherichia coli TOP10. These methods detected pathogenic strains from apple orchards. Pss might exist in an apple tree during ice injury, and it secretes a toxin that makes leaves yellow and cause canker symptoms. Until now, Korea has not developed antibiotics targeting Pss. Therefore, it is necessary to develop effective disease control to combat Pss in apple orchards. Pathogenicity test on apple leaves and stems showed canker symptoms. The pathogenic bacterium was re-isolated from symptomatic plant tissue and confirmed as original isolates by 16S rRNA. Repetitive element sequence-based PCR and enterobacterial repetitive intergenic consensus PCR primers revealed different genetic profiles within P. syringae pathovars. High antibiotic susceptibility results showed the misreading of mRNA caused by streptomycin and oxytetracycline.

Analysis of Algal Response Effects Using Nutrient Enrichment Bioassays in Shingu Reservoir (신구저수지 식물플랑크톤성장에 대한 제한 영양염 분석)

  • Lee, Jae-Yon;Lee, Sang-Jae;Lee, Jae-Hoon;Bae, Dae-Yeul;Choi, Ji-Woong;Hwang, Soon-Jin;An, Kwang-Guk
    • Korean Journal of Ecology and Environment
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    • v.41 no.spc
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    • pp.35-41
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    • 2008
  • Nutrient Enrichment Bioassays (NEBs) were conducted in the laboratory during June $22{\sim}28$, 2006 in order to determine primary limiting factor on the phytoplankton growth. For the NEBs, the water was sampled using a 10L polyethylene-lined container and dispensed into 2.5L container in the laboratory. The algal growths response in the control (C) and three treatments of phosphorus (P), 2-fold phosphorus (2P), and nitrate nitrogen $(NO_3-N)$ were monitored during 7 days. In the cubitainers which were spiked as P (T1) and 2P (T2) Chl-${\alpha}$ concentrations were decreased during the test period and the final concentrations was low than initial values. However, Chl-${\alpha}$ in the cubitainers which were spiked as $NO_3$(T3) and $P+NO_3$(T4) showed significant increases compared to the initial values, indicating that in the short-term experiments, nitrogen seemed to be a primary limiting nutrient during the periods of NEBs experiment. Long-term ambient nutrient data of TP and TN, and TN:TP mass ratios, however, showed a potential phosphorus limitation on phytoplankton growth and previous other researchers showed a variations of limiting nutrients by nitrogen or phosphorus depending on the seasons sampled and locations. In this study nitrogen as primary limiting nutrient in the NEBs seem to be an seasonal effect rather than the consistent nitrogen limitation.