• Title/Summary/Keyword: 생물검정법

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Improved Soil Application Bioassay for Efficient Development of Natural Pre-emergence Herbicides (토양처리용 천연물제초제 개발을 위한 생물검정법 개선)

  • Kim, Jae-Deog;Hwang, Hyun-Jin;Seo, Bo-Ram;Choi, Jung-Sup;Kim, Jin-Seog
    • Korean Journal of Weed Science
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    • v.31 no.3
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    • pp.229-239
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    • 2011
  • This study was carried out to establish an improved bioassay system on the side of practicality, pre-emergence bioassay which is more effective in developing soil application natural herbicides. A miniaturized method which have a 50 cm2 of soil surface area and was efficient by 7 times compared to the existing soil application assay ($350cm^2$ of soil surface area) was established, in which four weed species (Echinochloa crus-galli, Digitaria sanguinalis, Aeschynomene indica, and Abutilon theophrasti) were planted and grown in greenhouse. This would be applicable when the amount of screening compound is much more than 50 mg. The initial application rate was desirable at $10,000{\mu}g\;mL^{-1}$. On the other hand, the 6 well plate assay which has 4 weed species in each well containing upland soil and could be conducted in growth chamber, was established. This assay was resulted in minimizing in level of 1/14 test volume and 1/14 amounts of test compound to the conventional method that has been used for screening of synthetic compounds in KRICT, and applicable for the small amount of test compound (less than 10 mg). Therefore, the improved bioassays established in this study would be helpful for a rapid and efficient development of soil application natural herbicides.

Detection Methods for Genetically Modified Soybeans (유전자변형 콩의 검정법)

  • Sohn, Seong-Han;Jeong, Soon-Il;Yoon, Mun-Sup;Kim, Tae-San;Park, Yong-Hwan;Kim, Young-Mi
    • Applied Biological Chemistry
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    • v.45 no.4
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    • pp.185-189
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    • 2002
  • Along with the worldwide rapid increase of the cultivation area and commercial production of genetically modified (GM) crops, the amount of GM grains imported to Korea has also been increasing. Roundup-Ready soybean (RRS) was introduced with 5-enolpyruvyl shikimate-3-photphate synthase (EPSPS) gene derived from Agrobacterium CP4 to confer the resistance to herbicide, glyphosate. In this study, we tried to develop PCR-based analytical method to detection the presence of RRS among non-GM soybeans. In order to detect RRS specifically, oligonucleotide primers were specifically designed based on the nucleotide sequence of EPSPS transgene. Qualitative PCR method was established and its specificity and accuracy were confirmed by analysing the nucleotide sequence of PCR DNA fragments. Bioassay was also conducted by spraying glyphosate at seedling stage. Survived individuals showed obvious resistance to Roundup Ready, however all of non-GM seedlings died in two weeks after spray. Conclusively, the highly selective detection systems for RRS were successfully established by both PCR using specific primers to EPSPS transgene and bioassay using the herbicide resistance of RRS. In addition to, the imported soybean showed to be mixed to several varieties regarding to 100-seed weight and hilum color.

Development of simultaneous detection method for living modified cotton varieties MON757, MON88702, COT67B, and GHB811 (유전자변형 면화 MON757, MON88702, COT67B, GHB811의 동시검출법 개발)

  • Il Ryong Kim;Min-A Seol;A-Mi Yoon;Jung Ro Lee;Wonkyun Choi
    • Korean Journal of Environmental Biology
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    • v.39 no.4
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    • pp.415-422
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    • 2021
  • Cotton is an important fiber crop, and its seeds are used as feed for dairy cattle. Crop biotechnology has been used to improve agronomic traits and quality in the agricultural industry. The frequent unintentional release of LM cotton into the environment in South Korea is attributed to the increased application of living modified (LM) cotton in food, feed, and processing industries. To identify and monitor the LM cotton, a method for detecting the approved LM cotton in South Korea is required. In this study, we developed a method for the simultaneous detection of four LM cotton varieties, MON757, MON88702, COT67B, and GHB811. The genetic information of each LM event was obtained from the European Commission-Joint Research Centre and Animal and Plant Quarantine Agency. We designed event-specific primers to develop a multiplex PCR method for LM cotton and confirmed the specific amplification. Using specificity assay, random reference material(RM) mixture analysis and limit of detection(LOD), we verified the accuracy and specificity of the multiplex PCR method. Our results demonstrate that the method enabled the detection of each event and validation of the specificity using other LM RMs. The efficiency of multiplex PCR was further verified using a random RM mixture. Based on the LOD, the method identified 25 ng of template DNA in a single reaction. In summary, we developed a multiplex PCR method for simultaneous detection of four LM cotton varieties, for possible application in LM volunteer analysis.

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.

Use of comet assay as a bioassay in marine organisms exposed to genotoxicants (유전독성물질로 오염된 해양생물의 생물검정법으로서 comet assay 이용)

  • Kim Gi-Beum;An Joon-Gun;Kim Jae-Won
    • Journal of Environmental Science International
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    • v.14 no.11
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    • pp.1071-1079
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    • 2005
  • Using single cell gel electrophoresis, DNA single strand breaks were determined in various marine organisms. DNA damage on fish blood cells was detected to know whether there was a difference between Incheon, Pohang, Masan, and Tongyeong as a control site. Tongyeong showed the lowest DNA damage among the study areas. Mussels, transplanted to Masan Bay for one month, revealed high DNA damage at sites with high economical activity. In two weeks exposure of polychaete to Incheon sediments, higher DNA damage was detected in the sediment adjacent to Incheon harbor than open sea. These results suggested that the marine organism from the polluted area revealed a relatively high DNA damage. In addition, these areas might be contaminated with genotoxic compounds and comet assay was useful as a bioassay to detect DNA damage in marine organisms.

Individual Bioequivalence Tests under 3 X 2 Design (3 X 2 교차설계법에서 개인 생물학적 동등성 검정)

  • Jung, Gyu-Jin;Lim, Nam-Kyoo;Park, Sang-Gue
    • The Korean Journal of Applied Statistics
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    • v.23 no.1
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    • pp.139-150
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    • 2010
  • In recent years, more generic drug products became available. The current regulation for assessing the bioequivalence of two drug formulations is based on the concept of average bioequivalence. This approach has been indicated to be insufficient for assessing switchability between two drug formulations and US FDA has adopted individual bioequivalence as one of the bioequivalence criterion since 2001. The US FDA recommends that individual bioequivalence be assessed based on $2\;{\times}\;4$ crossover design, while a $2\;{\times}\;3$ crossover design may be used as an alternative design to reduce the length and cost of the study. In this paper, a statistical procedure for assessment of individual bioequivalence under $3\;{\times}\;2$ crossover designs is proposed and some statistical points are discussed with $2\;{\times}\;3$ crossover design and $2\;{\times}\;3$ extra-reference design through simulation studies.

Examining the Interrelation of Total, Soluble, and Bioavailable Metals in the Sediments of Urban Artificial Lakes (도심인공호 퇴적물의 총중금속, 용존중금속, 생물이용성 중금속의 연관성 규명)

  • Baek, Yong-Wook;An, Youn-Joo
    • Korean Journal of Ecology and Environment
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    • v.41 no.1
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    • pp.66-72
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    • 2008
  • Total metals, soluble metals, and bioavailable metals were monitored at the sediments of urban lakes located in Seoul, Korea during spring season 2006. The metals measured were zinc, arsenic, chromium, copper, nickel, and cadmium, which are known to be toxic to human health and ecosystems. The main sources of heavy metals in the lakes were urban runoff and atmospheric deposition associated with air pollution in urban areas. Extraction by using a weak electrolyte solution (0.1 M $Ca(NO_3)_2$) was used to predict bioavailability of the metals. Among the six heavy metals studied, copper was the most bioavailable, based the weak electrolyte extraction techniques. Since metal toxicity is related to metal bioavailability, the results were consistent with the high ecotoxicity of copper, compared to other heavy metals. Overall results suggest that there was no direct relationship between total and bioavailable metal concentration, although zinc, copper and cadmium show some relationships.

Investigation of brassinosteroid substances in the shoots of japonica type Korean rice (일반계 벼의 brassinosteroid활성물질 검색)

  • Hyun, Kyu-Hawn;Park, Keun-Hyung
    • Applied Biological Chemistry
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    • v.33 no.4
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    • pp.307-314
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    • 1990
  • Brassinosteroid-like substances in two japonica type Korean rices were investigated. The extracts from the shoots at the maximum tillering stage were purified by solvent fractionation, silica gel adsorption chromatography, Sephadex LH-20 chromatography, charcoal adsorption chromatography, preparative TLC, Bondesil chromatography and HPLC of normal phase and reverse phase, successively. Biological activity of each purification step were monitored by the rice lamina inclination test. Two cultivars tested in this experiment produced brassinosteroids and endogenous brassinosteroids showing similiarity between two cultivars.

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A Study on the Determination of Residual Antibiotics and synthetic antibacterial agents in Meat(I) -Microbiological Assay- (식육중의 잔류 항생.항균제의 검정에 관한 연구(I) -생물학적 검정법-)

  • 류재천;송윤선;박종세;장준식;신보승언
    • Journal of Food Hygiene and Safety
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    • v.8 no.1
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    • pp.1-8
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    • 1993
  • 육류중에 잔류하는 항생물질 및 항균제를 검출하기 위해 본 실험에서는 3종의 균주 Bacillus subtilis ATCC 6633, Micrococcus luteus ATCC 9341, BAcillus cereus var. mycoides ATCC 11778을 사용하여 실험하였다. 시료의 clean-up은 항생·항균제의 물리화학적 성질을 고려하여 우선 McIlvaine buffer를 가하여 homogenize하고 hexane으로 defatting 시킨 후, chloroform으로 추출한 액과 Sep-Pak C18과 Bakerbond SPE carboxylic acid column에 흡착시킨 후 추출한 액을 각각 시험용액 A, B, C,로 하였다. 각 test solution을 paper disk를 사용하여 함균 배지에 올려놓고 overnight culture 후 inhibition pattern을 통해 여러 종류의 항생·항균제를 계통적으로 검출하였다. 본 실험에서 macrolide계와 tetracycline계 등은 0.1ppm 이하의 detection limit을 보였으며, penicillinrP는 0.001ppm이하의 높은 detection limit을 나타내므로서 시료중에 잔류하는 극미량까지도 검출할 수 있었다. 본 방법은 식육중의 잔류 항생·항균제를 동시에 간단하게 계통적으로 분류하는데 있어 좋은 방법이라고 생각되며 항생·항균제의 체계적인 1차 screening 수단으로서 유용한 방법이라 사료된다.

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Isolating Entomopathogenic Nematode in South Korea (남한 토양에서 곤충병원성 선충의 분리)

  • 한상미;한명세
    • The Korean Journal of Ecology
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    • v.22 no.5
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    • pp.255-263
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    • 1999
  • Entomopathogenic nematodes were isolated through the investigation of soil samples from various biotopes in south Korea, the efficiency of isolation for highly pathogenic nematodes to silkworms (Bombyx mori) was as high as 28 %. Twenty-eight strains of nematodes, selected among 100 samples by silkworms were confirmed the pathogenicity, multiplicity, and tolerance against various condition of preservation. Pathogenicity of the nematode isolates to agricultural and environmental pests such as Calliphora vomitoria, Pseufaletia separata, Palomena angulosa, and Melolontha incana were high. Mortality was varied from 20 to 100% by the pest insects and nematode strains. The high detectablity of entomopathogenic nematodes resulted from the methods of collection for soil samples within 10 cm depth after eliminating dried soil surface and the use of silkworm trap. High population of entomopathogenic nematodes represented the strong activity and broad action radius in the environment. Most of the nematode isolates were successfully cultured on the silkworm host as well as on artificial media, and proved their potential for the use of biological control agent.

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