• Title/Summary/Keyword: 생물검정법

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Monoclonal antibody production for CP4 EPSPS detection assays (CP4 EPSPS 검출을 위한 단클론 항체 생산)

  • A-Mi Yoon;Il Ryong Kim;Wonkyun Choi
    • Korean Journal of Environmental Biology
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    • v.39 no.4
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    • pp.445-451
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    • 2021
  • In this study, we described the production of an antibody to living modified organisms (LMOs) containing the gene encoding for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium tumefaciens strain CP4 EPSPS provides resistance to the herbicide glyphosate (N- (phosphonomethyl) glycine). These LMOs were approved and have recently been used in the feed, food production, and processing industries in South Korea. Highly efficient monoclonal antibody (mAb) production is crucial for developing assays that enable the proper detection and quantification of the CP4 EPSPS protein in LMOs. This study describes the purification and characterization of recombinant CP4 EPSPS protein in E. coli BL21 (DE3) based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrixassisted laser desorption/ionization time-of-flight mass spectrometry. The production of mAbs was undertaken based on the standard operating procedure of Abclon, Inc.(South Korea), and the purity of the mAbs was assessed using SDS-PAGE. The following five mAb clones were produced: 2F2, 4B9, 6C11, 10A9, and 10G9. To verify the efficiency and specificity of the five developed mAbs, we performed Western blotting analysis using the LM (living modified) cotton crude extracts. All mAbs could detect the CP4 EPSPS protein in the LM cotton traits MON1445 and MON88913 with high specificity, but not in any other LM cottons or non-LM cottons. These data indicate that these five mAbs to CP4 EPSPS could be successfully used for the further development of antibody-based detection methods to target CP4 EPSPS protein in LMOs.

Screening of Antifungal Activities of Medicinal Plants for the Control of Turfgrass Fungal Disease (잔디 병해 방제를 위한 약용식물의 항균작용 탐색)

  • Kang, Jae Young;Kim, Dae Ho;Lee, Dong Gu;Kim, In Seob;Jeon, Min Goo;Lee, Jae Deuk;Kim, Ik Hwi;Lee, Sanghyun
    • Weed & Turfgrass Science
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    • v.2 no.1
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    • pp.70-75
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    • 2013
  • Seven medicinal plant extracts were tested for antifungal activities against six species of the major turfgrass pathogenic fungi (Colletotrichum graminicola, Pythium spp., Rhizoctonia cerealis, Rhizoctonia solani AG1-1, Rhizoctonia solani AG2-2, and Sclerotinia homoeocarpa) using paper disk diffusion method. Three medicinal plant extracts, including Pinus densiflora showed antifungal activities. In suppression of mycelium growth test, on medium adding P. densiflora extract showed that inhibition rate of mycelium growth were above 80% in 10 mg/10 ml concentration of the extract. The inhibition rate of Pythium spp. was 100% and C. graminicola was 84.3% in 10 mg/10 ml concentrations of P. densiflora extract, respectively. In particularly, the inhibition rate of Pythium spp. was 89.5% in 2 mg/10 ml concentrations of P. densiflora extract. As a result, P. densiflora extract showed high antifungal activity to Pythium spp. and C. graminicola of the turfgrass pathogen in in vitro test.

Biological Activities of Essential Oil from Chamaecyparis obtusa (편백(Chamaecyparis obtusa) 정유의 항균, 항염, 항산화 효과)

  • Ahn Jeung-Youb;Lee Sung-suk;Kang Ha-young
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.4 s.48
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    • pp.503-507
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    • 2004
  • The essential oil from Chamaecyparis obtusa was investigated for biological activities in anti-oxidative, anti-inflammation and antibacterial method, respectively. The Growth inhibitory effect of C. obtusa oil on the bacteria was evaluated with MIC (minimum inhibitory concentration), $IC_{50}\;(50\%$ inhibitory concentration), and paper disc method. Two kinds of gram positive strains and two kinds of gram negative strains were used in this study. Gram positive strains were B. subtilis and S. aureus. and Gram negative strains were E. coli and P. aeruginosa. Gram positive strains showed much more intensive effect than gram negative strains. Anti-oxidative effect was investigated with DPPH (1,1-diphenyl-2-picrylhidrazyl) in methanol based and $IC_{50}\;was\;0.78\%.$ Our results suggest that the essential oil from Chamaecyparis obtusa has effects on anti-bacterial, anti- oxidative and anti-inflammation in in vitro and in uiuo. Then this material could be expect synergic effect with other candidated extracts and oils.

Bean common mosaic virus and Peanut mottle virus isolated from Peanut in Korea (땅콩(Arachis hypogaea)에서 분리한 Bean common mosaic virus와 Peanut mottle virus)

  • Koo, Dong-Jin;Shin, Hye-Young;Sung, Jung-Hyun;Kang, Dong-Kyon;Chang, Moo-Ung
    • Research in Plant Disease
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    • v.8 no.2
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    • pp.92-100
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    • 2002
  • For the survey of viruses infected in peanut cultivated in Korea, peanut seeds and leaves showing viral symptoms were collected from their growing areas. Typical symptoms on virus infected peanut leaves including mosaic, mottle with necrosis, yellowing, stripe or vein banding and stunts were observed. Two viruses isolated from the naturally infected peanuts were identified as Bean common mosaic virus(BCMV-PSt) and Peanut mottle virus(PeMoV) by their host range, immunosorbent elcetron microscopy(ISEM), direct immuno staining assay(DISA), RT-PCR, and intracellural symptoms. Direct negative staining method by electron microscope showed filamentous particles of about 780 m in length as well as inclusion bodies. In ultrathin sections of BCMV-PSt and PeMoV infected tissues, cytoplasmic cylindrical inclusions as well as filamentous virus particles were observed in the cytoplasm of parenchyma cells. ISEM revealed filamentous particles strongly decorated with antiserums of BCMV-PSt and PeMoV Peanut seeds were stained with BCMV-PSt and PeMoV antisera indicating the possibility of seed transmission far these viruses. Seedlings germinated from peanut seeds which reacted with antiserums of BCMV-PSt by DISA showed mild mottle or stripe symptoms while mosaic and necrotic mottle symptoms were observed for PeMoV-positive seedlings. Filamentous particles were strongly decorated with each antiserum under ISEM observation. BCMV-PSt coat protein gene of about 1.2 Kbp was amplified by RT-PCR. Altogether these results indicate that BCMV-PSt is the most prevalent virus infecting peanut in Korea.

Insecticide Resistance Monitoring of Bemisia tabaci (Hemiptera: Aleyrodidae) in Korea (전국 담배가루이 약제 저항성 조사)

  • Kim, Sanghyeon;Kim, Sung Jin;Cho, Susie;Lee, Si Hyeock
    • Korean journal of applied entomology
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    • v.60 no.2
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    • pp.167-173
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    • 2021
  • Sweet potato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is an insect pest with sucking mouth part and globally attacks diverse crops and vegetables. Since first reported in 1998, it is now widely spread in many regions in Korea. Due to insecticide resistance, it is necessary for optimal control of the whiteflies to select effective insecticides with precise insecticide resistance monitoring. In this study, B. tabaci individuals were collected from 12 regions in 7 provinces of Korea from June to September in 2020. Using these field populations, insecticide resistance levels were monitored using a residual contact bioassay along with molecular markers. Bioassay results revealed that B. tabaci possessed high levels of resistance to five insecticides exhibiting different modes of action: dinotefuran, spinosad, emamectin benzoate, chlorfenapyr, and bifenthrin. In addition, quantitative sequencing in target sites of organophosphate and pyrethroid insecticides revealed that point mutations reached to saturated or near-saturated levels across the country. This suggests that insecticide resistance management is required for effective control of B. tabaci populations in Korea.

Review of the study on the surfactant-induced foliar uptake of pesticide (계면활성제에 의해 유도되는 농약의 엽면 침투성 연구 현황)

  • Yu, Ju-Hyun;Cho, Kwang-Yun;Kim, Jeong-Han
    • The Korean Journal of Pesticide Science
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    • v.6 no.1
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    • pp.16-24
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    • 2002
  • Research trends in the measurement of foliar uptake of pesticides and the recently proposed action mechanism of the surfactant-induced uptake of pesticides were reviewed with the related reports and studies. Major techniques used in those fields are bioassay, radiotracer techniques with leaves or cuticular membrane. Recently, a new method using Congo Red as a tracer was proposed. The limiting factor in the pesticides uptake into leaves is the waxy layer which consists of the epicuticular and cuticular wax. Physico-chemical parameters such as molar volume, water solubility and partition coefficient of pesticides have limited influences on the pesticide uptake into leaves. Polydisperse ethoxylated fatty alcohol surfactants are well known as the good activator for many pesticides. It is now generally agreed that uptake activation is not related to the intrinsic surface active properties of surfactants such as surface activity, solvent property, humectancy and critical micelle concentration. Recent studies using ESR-spectroscopy revealed that the surfactants have an unspecific plasticising effect on the molecular structure of the wax and cuticular matrix, leading to increased mobilities of pesticides. Penetration of surfactants into waxy layer altered the pesticide mobility in wax and the partition coefficient of pesticide, and then the pesticides penetration into leaves was enhanced temporally. The enhancing effect of surfactant could be significantly different depending on the carbon number of aliphatic moiety and the number of ethoxy group in polyoxyethylene chain of surfactants. It is suggested that the rate of penetration of surfactants should have a significant relationship with the rate of penetration of pesticides.

Development of sequential sampling plan for Frankliniella occidentalis in greenhouse pepper (고추 온실에서 꽃노랑총채벌레의 축차표본조사법 개발)

  • SoEun Eom;Taechul Park;Kimoon Son;Jung-Joon Park
    • Korean Journal of Environmental Biology
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    • v.40 no.2
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    • pp.164-171
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    • 2022
  • Frankliniella occidentalis is an invasive pest insect, which affects over 500 different species of host plants and transmits viruses (tomato spotted wilt virus; TSWV). Despite their efficiency in controling insect pests, pesticides are limited by residence, cost and environmental burden. Therefore, a fixed-precision level sampling plan was developed. The sampling method for F. occidentalis adults in pepper greenhouses consists of spatial distribution analysis, sampling stop line, and control decision making. For sampling, the plant was divided into the upper part(180 cm above ground), middle part (120-160 cm above ground), and lower part (70-110 cm above ground). Through ANCOVA, the P values of intercept and slope were estimated to be 0.94 and 0.87, respectively, which meant there were no significant differences between values of all the levels of the pepper plant. In spatial distribution analysis, the coefficients were derived from Taylor's power law (TPL) at pooling data of each level in the plant, based on the 3-flowers sampling unit. F. occidentalis adults showed aggregated distribution in greenhouse peppers. TPL coefficients were used to develop a fixed-precision sampling stop line. For control decision making, the pre-referred action thresholds were set at 3 and 18. With two action thresholds, Nmax values were calculated at 97 and 1149, respectively. Using the Resampling Validation for Sampling Program (RVSP) and the results gained from the greenhouses, the simulated validation of our sampling method showed a reasonable level of precision.

Study on the Screening System of Organic Resources for Agricultural Utilization (유기성 자원의 농업적 활용을 위한 선별체계 연구)

  • Lim, Dong-Kyu;Lee, Seung-Hwan;Kwon, Soon-Ik;So, Kyu-Ho;Sung, Ki-Suk;Koh, Mun-Hwan;Lee, Jeong-Taek
    • Korean Journal of Soil Science and Fertilizer
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    • v.38 no.2
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    • pp.92-100
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    • 2005
  • This study was conducted to find suitable methods for screening organic resources useful for compost. Twenty-seven industrial and domestic sludges were collected from various cities and industrial areas. Contents of organic matters in the sludges were in the range of 79.3-98.0%, and the contents were much higher than the regulation level (60%) for raw materials of compost. Contents of total nitrogen were in the range of 0.8-2.6%. Contents of Fe and Al were very high. Content of HEM was highest in textile sludge ($257mg\;kg^{-1}$) and the contents in the others were in the range of $12.6-90.3mg\;kg^{-1}$. Content of PAHs was lowest in food sludge ($739.1{\mu}g\;kg^{-1}$ and pulp-mill sludge had the highest PAHs content ($3461.8{\mu}g\;kg^{-1}$). $Microtox^{(R)}$ $EC_{50}$ values were higher in the sludges which were classified as a possible material in composting after analysis and investigation. Lettuce root elongation and $EC_{50}$ values were relatively lower in pulp-mill sludge, sewage sludge 3 (Large city), food sludge and leather sludge. Therefore, mineral nutrients, heavy metals, organic compounds (HEM, PAHs, PCBs), and bioassay ($Microtox^{(R)}$ $EC_{50}$, Relative root elongation test) are recommended to be included in the screening system of raw material of compost in addition to the current regulation with organic matter and 8 heavy metals.

Decomposition Characteristics of Fungicides(Benomyl) using a Design of Experiment(DOE) in an E-beam Process and Acute Toxicity Assessment (전자빔 공정에서 실험계획법을 이용한 살균제 Benomyl의 제거특성 및 독성평가)

  • Yu, Seung-Ho;Cho, Il-Hyoung;Chang, Soon-Woong;Lee, Si-Jin;Chun, Suk-Young;Kim, Han-Lae
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.9
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    • pp.955-960
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    • 2008
  • We investigated and estimated at the characteristics of decomposition and mineralization of benomyl using a design of experiment(DOE) based on the general factorial design in an E-beam process, and also the main factors(variables) with benomyl concentration(X$_1$) and E-beam irradiation(X$_2$) which consisted of 5 levels in each factor was set up to estimate the prediction model and the optimization conditions. At frist, the benomyl in all treatment combinations except 17 and 18 trials was almost degraded and the difference in the decomposition of benomyl in the 3 blocks was not significant(p > 0.05, one-way ANOVA). However, the % of benomyl mineralization was 46%(block 1), 36.7%(block 2) and 22%(block 3) and showed the significant difference of the % that between each block(p < 0.05). The linear regression equations of benomyl mineralization in each block were also estimated as followed; block 1(Y$_1$ = 0.024X$_1$ + 34.1(R$^2$ = 0.929)), block 2(Y$_2$ = 0.026X$_2$ + 23.1(R$^2$ = 0.976)) and block 3(Y$_3$ = 0.034X$_3$ + 6.2(R$^2$ = 0.98)). The normality of benomyl mineralization obtained from Anderson-Darling test in all treatment conditions was satisfied(p > 0.05). The results of prediction model and optimization point using the canonical analysis in order to obtain the optimal operation conditions were Y = 39.96 - 9.36X$_1$ + 0.03X$_2$ - 10.67X$_1{^2}$ - 0.001X$_2{^2}$ + 0.011X$_1$X$_2$(R$^2$ = 96.3%, Adjusted R$^2$ = 94.8%) and 57.3% at 0.55 mg/L and 950 Gy, respectively. A Microtox test using V. fischeri showed that the toxicity, expressed as the inhibition(%), was reduced almost completely after an E-beam irradiation, whereas the inhibition(%) for 0.5 mg/L, 1 mg/L and 1.5 mg/L was 10.25%, 20.14% and 26.2% in the initial reactions in the absence of an E-beam illumination.

Mapping and Race Specific Reaction of the Resistance Gene Pi45(t) in Rice (벼 도열병 저항성 유전자 Pi45(t)의 균계 특이적 반응과 고밀도지도 작성)

  • Kim, Dong-Min;Ju, Hong-Guang;Yang, Paul;Han, Seong-Sook;Roh, Jae-Hwan;Ahn, Sang-Nag
    • Korean Journal of Breeding Science
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    • v.43 no.1
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    • pp.42-49
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    • 2011
  • QTL analysis for blast resistance was carried out using 140 $BC_3F_3$ lines derived from a cross between Ilpum as a recurrent parent and Moroberekan as a donor parent. 140 $BC_3F_3$ lines with the parents were inoculated with nine blast isolates. To identify QTLs for resistance to nine blast isolates, 134 SSR markers showing polymorphisms between the parents were genotyped for the 140 $BC_3F_3$ lines. A total of 17 resistance QTLs to nine isolates were detected on chromosomes 2, 3, 4, 6, 7, 9 and 10. The phenotypic variance explained by each QTL ranged from 8.2% to 26.4%. The Moroberekan alleles contributed the positive effect at these 17 QTL loci. In a previous study, the QTL, Pi45(t) for durable resistance to blast was identified using a sequential planting method. To know the relationship between Pi45(t) and the isolate-specific resistance gene, an $F_2$ population was developed from a cross between Ilpum and an introgression line harboring Pi45(t). $F_3$ lines segregating for the Pi45(t) were inoculated to three isolates. $F_3$ lines from the $F_2$ plants with the Moroberekan segment at the target region showed resistance to two isolates. This result seems to indicate that the Pi45(t) and the isolate-specific resistance gene are tightly linked or the resistance is controlled by the same gene(s). The markers linked to genes controlling blast resistance would be useful in developing blast resistance lines in the breeding program.