• Title/Summary/Keyword: Bioassays

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Gibberellin A4 Producted by Fusarium solani Isolated from the Roots of Suaeda japonica Makino (칠면초의 뿌리로부터 분리된 Fusarium solani에 의해 생산된 지베렐린 A4)

  • Seo, Yeonggyo;You, Young-Hyun;Yoon, Hyeokjun;Kang, Sang-Mo;Kim, Hyun;Kim, Miae;Kim, Changmu;Lee, In-Jung;Kim, Jong-Guk
    • Journal of Life Science
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    • v.22 no.12
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    • pp.1718-1723
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    • 2012
  • Ten endophytic fungi with different colony morphologies were isolated from the roots of Suaeda japonica Makino growing naturally in Suncheon Bay. Plant growth promotion was verified by treating waito-c rice seedlings with culture filtrates from the endophytic fungi. The bioassays showed that the Sj/7/4 fungal strain induced effective growth promotion in the seedlings. The gibberellins (GA) produced by fungal strain Sj/7/4 were analyzed by gas chromatography /mass spectroscopy with selected ion monitoring (GC/MS SIM). The culture filtrate of Sj/7/4 fungal strain was confirmed to contain $GA_4$ through quantitative analysis. The Sj/7/4 fungal strain was identified to determine the internal transcribed spacer (ITS) regions with universal primers ITS-1 and ITS-4 by using polymerase chain reactions (PCR). Molecular analysis of the Sj/7/4 fungal strain showed high similarity to Fusarium solani. The Sj/7/4 fungal strain isolated from the root of S. japonica was therefore designated as F. solani Sj/7/4.

Transgenic Rice Expressing Snowdrop Lectin (Galanthus nivalis agglutinin; GNA) Shows Resistance to Rice Brown Planthopper (Nilaparvata lugens Stål) (Snowdrop lectin (Galanthus nivalis agglutinin: GNA) 유전자 도입에 의한 벼멸구 저항성 형질전환 벼 개발)

  • Lee, Soo In;Yoon, In Sun;Kim, Jin A;Hong, Joon Ki;Park, Beom-Seok;Lee, Yeon-Hee
    • Journal of Life Science
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    • v.22 no.12
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    • pp.1614-1620
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    • 2012
  • Transgenic rice plants with increased resistance to rice brown planthopper (Nilaparvata lugens St${\aa}$l) were generated by particle bombardment-mediated transformation of plants with a gene encoding snowdrop lectin (Galanthus nivalis agglutinin; GNA) under control of the rice Rubisco small subunit (rbcS) promoter.. A large number of transgenic rice plants containing the GNA gene were generated. The integration, expression, and inheritance of this gene in the $R_1$ and $R_2$ generations were demonstrated by Southern and western blot analyses. The plants contained one to five copies of the transgene. The GNA protein comprised approximately 0.01-2.0% of total soluble protein in the $R_1$ and $R_2$ transgenic plants. Insect bioassays and feeding studies showed that the GNA protein expressed in the $R_2$ transgenic rice plants reduced the survival of brown planthoppers. The introduction of GNA into rice plants therefore can help to control insect pests.

Replacement of the in vivo Bioassay for Erythropoietin with the in vitro Bioassay (Erythropoietin in vivo 시험법의 in vitro 대체 시험법 확립)

  • 백상훈;김진만;권기성;박송용;허재욱
    • KSBB Journal
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    • v.18 no.4
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    • pp.255-260
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    • 2003
  • In vivo bioassays for biological medicines have been considered final resort to unequivocally assess the biological activities for them because there are some cases in which the biological activities obtained from in vivo bioassay and in vitro bioassay quite differ each other. The in vivo biological activity of EPO depends on its sialic acid contents which confer microheterogeneity-isoforms to this protein. We have devise a method which consists of a in vitro bioassay using BaF3 cell line and a capillary zone electrophoresis (CZE) for the measurement of the EPO isoform distribution. The biological activity of EPO obtained using in vitro bioassay with BaF3 cell line showed good correlation (C.V.(%) 7.34, 5.85, 8,16, 8.08, 8.8) to EPO content measured either spectrophotometric assay (A280 0.1 % =0.743) or radio immunoassay. The assay validation results of in vitro bioassay with 3 lot of in house EPO showed good results to EPO content measured either in vivo assay or radio immunoassay. and also showed good results the robustness of our method in terms of precision, accuracy, repeatability. The isoform distribution for EPO-BRP (1 : 1 mixture of epoetin-${\alpha}$ and epoetin-${\beta}$, European Pharmacopoeia) by CZE method resulted in isoform 2 through isoform 8. The major peaks in electrophoregram were composed of isoform 3 through 7. Our recombinant EPO (epoetin-${\alpha}$) having equivalent in vivo biological activity showed the isoform distribution of isoform 3 through 9. The major peaks consisted of isoform 4 through 8. The peak area of isoform 4 was always smaller than that of isoform 5. The preparations of recombinant epoetin-${\alpha}$ with lower in vivo biological activity than EPO-BRP showed the isoform 2 through 8 in their electrophoregrams whose major peaks consisted of the isoform 3 through 7. The peak area of isoform 4 was larger than that of isoform 5.

Antioxidant and anti-inflammatory activity of extracts from red beet (Beta vulagaris) root (레드 비트 뿌리 추출물의 항산화 및 항염증 효과)

  • Yi, Mi-Ran;Kang, Chang-Hee;Bu, Hee-Jung
    • Food Science and Preservation
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    • v.24 no.3
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    • pp.413-420
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    • 2017
  • This study was designed to examine the in vitro antioxidant and anti-inflammatory effects of red beet (Beta vulagaris) root. Red beet root was extracted using 70% ethanol and then fractionated sequentially with n-hexane, ethyl acetate and butanol. Antioxidative ability was evaluated by bioassays using total polyphenol contents and ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid diammonium salt) radical scavenging activity. Ethyl acetate fraction of red beet root was best on total polyphenol contents ($37.02{\pm}0.37mg\;GAE/g$) and ABTS radical scavenging effects ($IC_{50}$ $42.9{\pm}9.5{\mu}g/mL$). For the anti-inflammatory activity in RAW264.7 cells, the hexane fraction showed the highest inflammatory effect. Dose response studies were performed to determine the inhibitory effect of hexane fraction of red beet root on pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. The hexane fraction of red beet root inhibited the NO and $PGE_2$ production and the protein level of iNOS and COX-2, and protein expression of pro-inflammatory cytokines ($TNF-{\alpha}$, IL-6 and $IL-1{\beta}$), in a dose-dependent manner. These results suggest that red beet root has considerable potential as a functional food ingredient with antioxidative and anti-inflammatory effects.

Antioxidant Activities of Korean Perilla Leaves (Perilla frutescens) by Various Cultivars (잎들깨 품종에 따른 추출물의 항산화 활성 및 생리활성 평가)

  • Hyun Young, Kim;HanGyeol, Lee;Hye Young, Seo;Woo Duck, Seo;Mi Ja, Lee;Seung-Yeob, Song;Jung In, Kim;June-Yeol, Choi
    • The Korean Journal of Food And Nutrition
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    • v.35 no.6
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    • pp.453-463
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    • 2022
  • This study aimed to investigate the biological activities, including polyphenolic content, flavonoid content, and antioxidant activities, of various cultivars of Korean perilla leaves. The results indicated that among nine cultivars (Namcheon dlggae, Saedora, Nulbora, Donggel 1, Donggell 2, Soim, Sangyeop, Somirang, and Saebom) of perilla leaves, the total polyphenolic content (gallic acid equivalent mg/g, GAE) was the highest in "Nulbora," while it was lowest in Namcheon dlggae. Moreover, flavonoid content in the extracts of nine cultivars leaves was in the range of 132.93~268.50 mg catechin equivalent/g sample. The antioxidant effects of the perilla leaves were determined using two different in vitro bioassays measuring DPPH and ABTS radical-scavenging activities. The results revealed that antioxidant activity was also higher in "Nulbora" compared with other cultivars. Xanthin-oxidase-inhibition activity ranged from 65.65% to 80.58%, with "Nulbora" exhibiting the highest activity, although the difference with other cultivars was not significant. "Nulbora" extracts reduced the expression of pro-inflammatory genes and several cytokines, including IL-6 activation induced by LPS in macrophages in the range of 100-50 ㎍/mL. These results suggest that extracts from perilla leaves can be used as bioactive and functional materials that could be important in industrial applications in the future.

An Optimal Standardized in vitro Bioassay to Evaluate Susceptibility of Green Peach Aphid, Myzus persicae (Sulzer)(Insecta: Hemoptera: Aphididae), to Aphicides (복숭아혹진딧물, Myzus persicae (Sulzer)(Insecta: Hemoptera: Aphididae), 살진딧물 최적 in vitro 살충력 검정 방법 확립)

  • Ka Hee Cho;Hyo Jung Kim;Young Cheol Kim
    • Korean journal of applied entomology
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    • v.62 no.3
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    • pp.139-147
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    • 2023
  • Leaf-spray in vitro bioassays appraise new aphicidal formulations for managing deleterious plant-feeding aphids. The formulation may utilize alternative and integrated strategies. However, leaf spraying even under controlled conditions may affect aphid reproduction and mortality. This study examines leaf spray applications for optimum and reproducible aphicidal results using tobacco leaves overlaid on cotton fabric or water agar surfaces. Infestation of the undersides of tobacco leaves with nymphs of green peach aphids was used in the assays. Spray distance and volume were optimized using water-sensitive paper to ascertain the best surface coverage. Overlays of the leaves on water agar caused less mortality and greater reproduction than the use of cotton fabric. The relative humidity of the insect-rearing chambers changed with the watering regime for the insect - rearing chambers with cotton fabric; 60% relative humidity was optimal. Relative humidity was not affected by the concentration of agar in the water agar chambers. Applications of the chemical aphicidal standard, Sulfoxaflor, under the optimized conditions exhibited similar times for lethality although the rate was faster with leaves on the cotton fabric than on water agar. These studies establish reproducible and sensitive techniques for assessing the lethality and effects on reproduction of potential aphicidal products.

Evaluation of Insecticidal and Antifeeding Activities of Eco-friendly Organic Insecticides Against Agricultural Insect Pests (농업해충에 대한 친환경유기농자재들의 살충력 및 섭식저해력 평가)

  • Kim, Yoo Hwa;Na, Young-Eun;Kim, Min Joon;Choi, Byung Ryul;Jo, Hyeong-Chan;Kim, Soon-Il
    • Korean journal of applied entomology
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    • v.54 no.2
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    • pp.99-109
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    • 2015
  • Insecticidal and antifeeding activities of 29 commercialized eco-friendly organic products for managing plant diseases and insect pests against Plutella xylostella larvae, Spodoptera exigua larvae, Frankliniella occidentalis adults, and Myzus persicae adults were tested using spraying and leaf dipping bioassays under laboratory conditions. Products containing 60% Sophora extract (EOIS) and mixtures (EOISm) with Sophora extract, Stemona japonica extract, Melia azedarach extract, and Nepeta cataria extract as well as mixtures (EOISc) with Sophora extract, Chenopodium ambrosioides extract, and Melia azedarach extract as active ingredients showed strong insecticidal activity at recommended concentration against P. xylostella larvae. At half concentration, their insecticidal activities were decreased under 50%. The EOIS gave good insecticidal activity against S. exigua larvae and also showed 85% and 95% insecticidal activity at 24 and 48 hours after treatment to F. occidentalis adults, respectively. For M. persicae adults, EOISm and mixtures (EOIR) containing rape seed extract, neem extract, and castar oil produced 93% and 68% insecticidal activity, but their activities did not be increased at double concentration. EOISm only showed 100% contact toxicity against M. persicae adults exposed to dipping leaves. Interestingly, the insecticidal activity of EOIR and EOICi (citronella oil and derris extract) against M. persicae adults was increased with exposed time and concentration. In addition, EOICe (cedar oil), EOIS, EOISm, EOISc, EOIM (microorganism), EOIR, EOIPe (plant extract), and EOIT (tea tree extract) gave strong antifeeding activity against S. exigua and P. xylostella larvae. EOIB, EOIBs, EOIM, EOICi, and EOIMc showed above 70% antifeeding activity to the lepidopteran larvae. These results indicate that mixtures containing 2 to 3 plant extracts with Sophora extract show good activities against insect pests, although the difference of insecticidal and antifeeding activities was produced depending on both a tested insect species and an active ingredient or concentration.

An Improved Method to Determine Corn (Zea mays L.) Plant Response to Glyphosate (Glyphosate에 대한 옥수수 반응의 개선된 검정방법)

  • Kim, Jin-Seog;Lee, Byung-Hoi;Kim, So-Hee;Min, Suk-Ki;Choi, Jung-Sup
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.57-62
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    • 2006
  • Several methods for determining the response of corn to glyphosate were investigated to provide a fast and reliable method for identifying glyphosate-resistant corn in vivo. Two bioassays were developed. One assay is named 'whole plant / leaf growth assay', in which the herbicide glyphosate is applied on the upper part of 3rd leaf and the growth of herbicide-untreated 4th leaf is measured at 3 day after treatment. in this assay, the leaf growth of conventional corn was inhibited in a dose dependent from 50 to $1600{\mu}g/mL$ of glyphosate and growth inhibition at $1600{\mu}g/mL$ was 55% of untreated control. The assay has the potential to be used especially in the case that the primary cause of glyphosate resistance is related with a reduction of the herbicide translocation. Another assay is named 'leaf segment / shikimate accumulation assay', in which the four excised leaf segments ($4{\times}4mm$) are placed in each well of a 48-well microtiter plate containing $200{\mu}L$ test solution and the amount of shikimate is determined after incubation for 24 h in continuous light at $25^{\circ}C$. In this assay, 0.33% sucrose added to basic test solution enhanced a shikimate accumulation by 3 to 4 times and the shikimate accumulation was linearly occurred from 2 to $8{\mu}g/mL$ of glyphosate, showing an improved response to the method described by Shaner et al. (2005). The leaf segment / shikimate accumulation assay is simple and robust and has the potential to be used as a high throughput assay in the case that the primary cause of glyphosate resistance is related with EPSPS, target site of the herbicide. Taken together, these two assays would be highly useful to initially select the lines obtained after transformation, to investigate the migration of glyphosate-resistant gene into other weeds and to detect a weedy glyphosate-resistant corn in field.

Phytotoxicity of Endophytic Fungi and Characterization of a Phytotoxin Isolated from Gliocladium catenulatum from Pinus densiflora (식물내생곰팡이의 제초활성 검정 및 소나무에서 분리한 Gliocladium catenulatum이 생산하는 제초활성 물질의 특성 규명)

  • Choi, Gyung-Ja;Park, Joong-Hyeop;Kim, Heung-Tae;Lee, Seon-Woo;Choi, Jung-Sup;Hong, Kyung-Sik;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Korean Journal of Mycology
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    • v.32 no.1
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    • pp.8-15
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    • 2004
  • This study was conducted to discover new phytotoxins which may be used as lead molecules for the development of new herbicides. A total of 187 endophytic fungi were isolated from 11 plant species, which were collected from 8 locations in Korea. Their herbicidal activities were screened in vivo by herbicidal and duckweed bioassays after they were cultured in potato dextrose broth and rice solid media. Both fermentation broth and solid culture extract of Gliocladium catenulatum F0006 isolated from red pine (Pinus densiflora) showed 70% herbicidal activity only against cocklebur (Xanthium strumarium) out of the 10 weeds tested. Solid culture extract of F0034 isolated from arrowroot (Pueraria thunbergiana) exhibited 20 to 100% herbicidal activities against all of the weeds. Especially, shattercane (Sorghum bicolor), barnyardgrass (Echinochloa crus-galli), large crabgrass (Digitaria sanguinalis), and fall pauicum (Panicum dichtomiflorum) were sensitive to the solid culture extract of F0034. In addition, solid culture extract of F0043 isolated from red pine displayed 20% to 70% herbicidal activities only against 5 grass species, but not against 5 broad-leaf plant species. On the other hand, as the results of duckweed assay, 8 fermentation broths showed 100% growth inhibitory activity at concentrations less than 5.0% of culture supernatants and 12 solid cultures had a potent inhibitory activity against duckweed growth. A toxic metabolite was purified from the solid cultures of G. catenulatum F0006 by repeated column chromatography and bioassay. It caused a phytotoxic syndrome only on cocklebur out of the 10 weeds tested; it completely killed cocklebur seedlings at $500\;{\mu}g/ml$ and showed 85% herbicidal activity against cocklebur at $100\;{\mu}g/ml$. The molecular weight of the toxic metabolite is 238 daltons and its structure determination is underway.

Using Trophic State Index (TSI) Values to Draw Inferences Regarding Phytoplankton Limiting Factors and Seston Composition from Routine Water Quality Monitoring Data (영양상태지수 (trophic state index)를 이용한 수체 내 식물플랑크톤 제한요인 및 seston조성의 유추)

  • Havens, Karl E
    • Korean Journal of Ecology and Environment
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    • v.33 no.3 s.91
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    • pp.187-196
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    • 2000
  • This paper describes a simple method that uses differences among Carlson's (1977) trophic state index (TSI) values based on total phosphorus (TP), chlorophyll a (CHL) and Secchi depth (SD) to draw inferences regarding the factors that are limiting to phytoplankton growth and the composition of lake seston. Examples are provided regarding seasonal and spatial patterns in a large subtropical lake (Lake Okeechobee, Florida, USA) and inter- and intra-lake variations from a multilake data set developed from published studies. Once an investigator has collected routine water quality data and established TSI values based on TP, CHL, and SD, a number of inferences can be made. Additional information can be provided where it also is possible to calculate a TSI based on total nitrogen (TN). Where TSI (CHL)<>TSI (SD), light attenuating particles are large (large filaments or colonies of algae), and the phytoplankton may be limited by zooplankton grazing. Other limiting conditions are inferred by different relationships between the TSI values. Results of this study indicate that the analysis is quite robust, and that it generally gives good agreement with conclusions based on more direct methods (e.g., nutrientaddition bioassays, zooplankton size data, zooplankton removal experiments). The TSI approach, when validated periodically with these more costly and time-intensive methods, provides an effective, low cost method for tracking long-term changes in pelagic structure and function with potential value in monitoring lake ecology and responses to management.

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