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Transgenic tobacco culture cells expressing spike protein gene of porcine epidemic diarrhea virus (돼지 유행성 설사병 바이러스 스파크 단백질 유전자 발현 형질전환 담배 배양세포)

  • Yang, Kyoung-Sil;Kim, Hyeon-Soo;Kwon, Suk-Yoon;Kwak, Sang-Soo;Lee, Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.35 no.1
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    • pp.87-94
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    • 2008
  • Porcine epidemic diarrhea virus (PEDV) is an infectious and highly contagious virus of swine. In order to develop the transgenic tobacco culture cells producing PEDV antigen protein, four vectors expressing PEDV spike protein (SP) gene under the control of a CaMV 35S promoter were constructed. Four fragments of the SP region of PEDV, SP1 (444 bp, 1487-1930 bp), SP2 (1.7 kb, 2300-3987 bp), SP3 (1.4 kb, 1559-2950 bp), and SP4 (2.6 kb, 9-2643 bp) were amplified by PCR and then C-MYC tag was fused to the end of each SP gene, respectively. These cassettes are inserted into the pCAMBIA2300 (named as 35S::SP1-M, 35S::SP2-M 35S::SP3-M, and 35S::SP4-M, respectively). Tobacco (cv. BY-2) cultured cells were transformed by co-cultivation with Agrobacterium tumefaciens harboring expression vector. We selected kanamycin-resistant calli and checked for the presence of the introduced SP gene using PCR, resulting 70% of them showed the foreign gene. We selected the lines with high-level expression of PEDV antigen protein based on dot blot analysis. Southern blot analysis confirmed that the PEDV SP gene was integrated into the genome of the tobacco cultured cells. Northern blot analysis showed that the introduced gene was highly expressed in transgenic cultured cells. Transgenic tobacco cultured cells-derived antigen induced immunogenicity in mice as determined by a plaque reduction neutralization assay. These results suggest that the vectors expressing PEDV spike protein gene in this study will be useful for the development of transgenic plants and cultured cells producing PEDV antigene protein.

Usefulness of Low Dose Oral Contrast Media in $^{18}F-FDG$ PET/CT ($^{18}F-FDG$ PET/CT에서 저용량 경구용 조영제의 유용성)

  • An Young-Sil;Yoon Joon-Kee;Hong Seon Pyo;Joh Chul-Woo;Yoon Seok-Nam
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.5
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    • pp.257-262
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    • 2006
  • Purpose: The standard protocol using large volume of oral contrast media may cause gastrointestinal discomfort and contrast-related artifacts in PET/CT. The aim of this study was to evaluate the usefulness of low dose oral contrast in $^{18}F-FDG$ PET/CT. Materials and Methods: We retrospectively reviewed the whole-body PET/CT images in a total of 435 patients. About 200 ml of oval contrast agent (barium sulfate) was administered immediately before injection of $^{18}F-FDG$. The FDG uptake of intestines was analyzed by visual and semi- quantitative method on transaxial, coronal and saggital planes. Results: Seventy (16%, 113 sites) of 435 images showed high FDG uptake (peak SUV > 4); 50 (74%, 84 sites) with diffuse and 20 (15%, 29 sites) with focal uptake. The most commonly delivered site of oral contrast media was small bowel (n=27, 39%). On PET/CT images, FDG uptake coexisted with oral contrast media in 26 patients (54%, 38 sites) with diffuse pattern and 9 (45%, 9 sites) with focal pattern, and by sites, those were 38 (45%) and 9 (31%), respectively. In small bowel regions, the proportion of coexistence reached as high as 61% (29/47 sites). A visual analysis of available non-attenuation corrected PET images of 27 matched regions revealed no contrast-related artifact. Conclusion: We concluded that the application of low dose contrast media could be helpful in the evaluation of abdominal uptake in the FDG PET/CT image.

A Study on Vegetation Structure and Soil Condition of $Bletilla$ $striata$ Population (자란($Bletilla$ $striata$) 개체군의 식생구조와 토양특성)

  • So, Soon-Ku;Song, Ho-Kyung
    • Korean Journal of Environment and Ecology
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    • v.26 no.2
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    • pp.210-218
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    • 2012
  • This study was carried out to analyze the vegetation properties, soil characteristics and ordination of Bletilla striata population in South Korea. The $Bletilla$ $striata$ population was classified into $Juniperus$ $rigida$ dominant population, $Rhododendron$ $yedoense$ var. $poukhanense$ dominant population, $Rhus$ $javanica$ dominant population, $Bletilla$ $striata$ typical population. $Bletilla$ $striata$ were mainly distributed along the coast of south-western regions of the Korean penninsula and it's population was located at an elevation of 4m to 40m. In the study sites, soil organic matter, total nitrogen, available phosphate, exchangeable potassium, exchangeable calcium, exchangeable magnesium, cation exchange capacity and soil pH were 2.38-6.70%, 0.09-0.27%, 1.81-6.85mg/kg, 1.08-1.72$cmol^+/kg$, 3.56-7.71$cmol^+/kg$, 1.52-3.21$cmol^+/kg$, 5.28-16.95$cmol^+/kg$ and 4.60-6.01 respectively. $Rhododendron$ $yedoense$ var. $poukhanense$ dominant population was found in the steep sloped area that has high percentage of organic matter, total nitrogen, cation exchange capacity and pH than other populations. $Rhus$ $javanica$ dominant population was found in the gentle sloped area that has less percentage of organic matter, total nitrogen, cation exchange capacity and pH. $Juniperus$ $rigida$ dominant population and $Bletilla$ $striata$ typical population were found in the medium sloped area that has medium percentage of organic matter, total nitrogen, cation exchange capacity and pH. Current status of $Bletilla$ $striata$ habitats is very vulnerable with local development constantly threatening the species' survival. Thus, concrete conservation plans to protect natural habitats should be set up as soon as possible.

Geochemical and Isotopic Study of the Onjeongri Granite in the Northern Gyeongsang Basin, Korea : Comparison with Cretaceous to Tertiary Granitic Rocks in the Other Part of the Gyeongsang Basin and the Inner Zone of Southwest Japan (경상분지 북부에 분포하는 온정리 화강암에 대한 암석화학적, 동위원소 지구화학적 연구 : 경상분지 다른 지역과 서남 일본 내대에 분포하는 백악기-제 3기 화강암류와의 비교 고찰)

  • 정창식;권성택;김정민;장병욱
    • The Journal of the Petrological Society of Korea
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    • v.7 no.2
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    • pp.77-97
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    • 1998
  • We analyzed geochemical and radiogenic isotope data to investigate the genesis and source characteristics of the Onjeongri granite in the northern part of the Gyeongsang Basin. Field observation and K-Ar ages confirm late Cretaceous intrusion (ca. 87 Ma) of the Onjeongri granite. The hornblende geobarometery gives less than 2 kbar for the emplacement pressure of the Onjeongri granite. Geochemical and isotopic compositions suggest that the Onjeongri granite was formed in a relatively immature arc system. $SiO_2$ contents show a negative linear relationship with initial $^{87}Sr/^{86}Sr$ ratios, and an apparent positive correlation with $^{207}Pb/^{204}Pb$ ratios, suggesting an incomplete mixing or assimilation. However, the isotopic data known for any exposed rocks of the study area do not fit as an endmember, implying that the contaminant might reside in the lower crust. A review of published isotopic ages, geochemical, and Sr and Nd isotopic data for the Cretaceous to Tertiary granites in the Gyeongsang Basin indicates the followings. 1) Granitic magmatism in the Gyeongsang Basin were episodic. 2) Granitic rocks in the basin were derived from young (< 0.9 Ga) lower crust, and their isotopic signatures reflect heterogeneous source region. Geochemical and isotopic signatures of granitic rocks in the basin are difficult to explain by upper crustal contamination. 3) Granites in the Gyeongsang Basin have closely related to those in the San in Belt of the Inner Zone of Southwest Japan in terms of age, petrography, and isotopic and geochemical composition. 4) Sr-Nd isotopic signatures of the Onjeongri granite are relatively primitive compared with granitic rocks in the other parts of the Gyeongsang Basin and in the Inner Zone of Southwest Japan.

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Studies on the Indigenous Vesicular-Arbuscular Mycorrhizal Fungi(VAMF) in Horticultural Crops Grown Under Greenhouse -III. Effects of the Indigenous VAMF Inoculation on the Early Growth and the Subsquent Growth after Transplanting of Greenhouse Grown Crops (시설원예(施設園藝) 작물(作物)에서 토착(土着) VA균근균(菌根菌)에 관한 연구(硏究) -III. 토착(土着) VA 균근균(菌根菌) 접종효과)

  • Sohn, Bo-Kyoon;Yang, Won-Mo;Kim, Kwang-Sik
    • Korean Journal of Soil Science and Fertilizer
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    • v.25 no.1
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    • pp.99-107
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    • 1992
  • Effects of the indigenous Vesicular-arbuscular mycurrhizal fungi(VAMF) on early growth response of greenhouse grown crops were experimented. This study was done to evaluate the benefit of indigenous VAMF inoculation on the early growth and the subsequent growth after transplanting of some crops such as cucumber, tomato, hot pepper, eggplant, and melon. Leaf area, shoot dry weight, and plant length of mycorrhizal greenhouse crops showed the tendency of significant or no significant increase over control plants receiving no inoculation. The levels of VA mycorrhizal colonization were increased with plant growth, and infection rates of horticultural crop except hot pepper around one week after transplanting were decreased, while that of 8 weeks after emergence of mycorrhizal seedlings were increased again and infected by around 50% at harvesting time. In spore densities in the rhizosphere soil of craps experimented, the number of spore ranged from $72.7{\pm}26.3$ to $100{\pm}10.3g^1$ on dried soil basis and high density showed in both cucumber and tomato. Total nitrogen contents in shoots were lower in the mycorrhizal plants than non-mycorrhizal one, whereas P uptake in mycorrhizal hot pepper and tomato were highly ramarkable. The K contents in the shoots of mycorrhizal cucumber and eggplent were highly enhanced. Inoculation of the indigenous VAMF enhanced shoot Ca and Mg in both tomoto and melon. The contents of Fe, Zn, Mn and Cu in shoots of mycorrhizal crops were higher than non-mycorrhizal plants and vice versa in case of eggplent. Inoculation of the indigenous VAMF to horticultural crops were effective for alleviation of transplanting shock, and pretransplanting infection improved subsequent growth by reducing the time required for establishment of a functional mycorrhizal symbiosis following transplanting.

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Antimicrobial Activity of GC-l00X against Major Food-Borne Pathogens and Detaching Effects of It against Escherichia coli O157:H7 on the surface of Tomatoes (GC-100X의 주요 식품위해 미생물에 대한 항균효과와 토마토 표면에 부착된 Escherichia coli O157:H7에 대한 세척 효과)

  • 박용호;권남훈;김소현;김지연;임지연;김준만;정우경;박건택;배원기
    • Journal of Food Hygiene and Safety
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    • v.17 no.1
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    • pp.36-44
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    • 2002
  • GC-l00X is non-corrosive alkaline ionic water (pH 12). It is composed of hydroxyl radicals and supplemented with xylitol. Its antimicrobial activity was examined against 6 major food-borne pathogens; Staphylococcus aureus FRI 913, Salmonella enterica serova Enteritidis ATCC 13076, S. enterica serova Typhimurium Korean isolate, Vibrio parahaemolyticus ATCC 17803, Escherichia coli O157:H7 ATCC 43894 and Pseudomonas aeruginosa KCTC 1637 at three different temperatures (4$^{\circ}C$, $25^{\circ}C$ and 36$^{\circ}C$) with or without an organic material (2% yeast extract), respectively. The antimicrobial activities showed over 4 log-reductions (1.0$\times$10$^4$CFU/ml reduction) against all pathogens reacted at 37$^{\circ}C$ for 3 hours in the absence of the organic material. The activities showed same results when GC-l00X was diluted with same volume of distilled water or standard hard water (CaCO$_3$300 ppm). Its antimicrobial activity was more effective and quicker in Gram-negative bacteria than Gram-positive bacteria. Its washing efficacy against E. coli O157:H7 exposed to the surfaces of tomatoes (grapes) was compared with that of the other sanitizers such as other kitchen synthetic detergent and 100-ppm chlorine water. For the toxicological evaluation of the sanitizers, viable counts of E. coli O157:H7 penetrated into the core of tomatoes after washing products were also compared. The result revealed that GC-100X stock solution and its 5% diluted solution had similar washing effects to 100-ppm chlorine water and more effective than the other kitchen synthetic detergent. This result indicated that GC- l00X had antimicrobial activity and no toxicological side effects, therefore, could be useful for a new sanitizer to use in flood safety and kitchen hygiene.

SURFACE CHARACTERISTICS AND BIOACTIVITY OF ANODICALLY OXIDIZED TITANIUM SURFACES (양극산화에 의한 티타늄 산화막의 표면 특성 및 생체 활성에 관한 연구)

  • Lee, Sang-Han;Cho, In-Ho
    • The Journal of Korean Academy of Prosthodontics
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    • v.45 no.1
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    • pp.85-97
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    • 2007
  • Statement of problem: Recently, anodic oxidation of cp-titanium is a popular method for treatment of titanium implant surfaces. It is a relatively easy process, and the thickness, structure, composition, and the microstructure of the oxide layer can be variably modified. Moreover the biological properties of the oxide layer can be controlled. Purpose: In this study, the roughness, microstructure, crystal structure of the variously treated groups (current, voltage, frequency, electrolyte, thermal treatment) were evaluated. And the specimens were soaked in simulated body fluid (SBF) to evaluate the effects of the surface characteristics and the oxide layers on the bioactivity of the specimens which were directly related to bone formation and integration. Materials and methods: Surface treatments consisted of either anodization or anodization followed thermal treatment. Specimens were divided into seven groups, depending on their anodizing treatment conditions: constant current mode (350V for group 2), constant voltage mode (155V for group 3), 60 Hz pulse series (230V for group 4, 300V for group 5), and 1000 Hz pulse series (400V for group 6, 460V for group 7). Non-treated native surfaces were used as controls (group 1). In addition, for the purpose of evaluating the effects of thermal treatment, each group was heat treated by elevating the temperature by $5^{\circ}C$ per minute until $600^{\circ}C$ for 1 hour, and then bench cured. Using scanning electron microscope (SEM), porous oxide layers were observed on treated surfaces. The crystal structures and phases of titania were identified by thin-film x-ray diffractmeter (TF-XRD). Atomic force microscope (AFM) was used for roughness measurement (Sa, Sq). To evaluate bioactivity of modified titanium surfaces, each group was soaked in SBF for 168 hours (1 week), and then changed surface characteristics were analyzed by SEM and TF-XRD. Results: On basis of our findings, we concluded the following results. 1. Most groups showed morphologically porous structures. Except group 2, all groups showed fine to coarse convex structures, and the groups with superior quantity of oxide products showed superior morphology. 2. As a result of combined anodization and thermal treatment, there were no effects on composition of crystalline structure. But, heat treatment influenced the quantity of formation of the oxide products (rutile / anatase). 3. Roughness decreased in the order of groups 7,5,2,3,6,4,1 and there was statistical difference between group 7 and the others (p<0.05), but group 7 did not show any bioactivity within a week. 4. In groups that implanted ions (Ca/P) on the oxide layer through current and voltage control, showed superior morphology, and oxide products, but did not express any bioactivity within a week. 5. In group 3, the oxide layer was uniformly organized with rutile, with almost no titanium peak. And there were abnormally more [101] orientations of rutile crystalline structure, and bonelike apatite formation could be seen around these crystalline structures. Conclusion: As a result of control of various factors in anodization (current, voltage, frequency, electrolytes, thermal treatment), the surface morphology, micro-porosity, the 2nd phase formation, crystalline structure, thickness of the oxide layer could be modified. And even more, the bioactivity of the specimens in vitro could be induced. Thus anodic oxidation can be considered as an excellent surface treatment method that will able to not only control the physical properties but enhance the biological characteristics of the oxide layer. Furthermore, it is recommended in near future animal research to prove these results.

Inhibitory Role of Polyamines in Dexamethasone-induced Apoptosis of Mouse Thymocytes (Dexamethasone에 의한 생쥐 흉선의 Apoptosis에서 Polyamine의 역할)

  • Choi, Sang-Hyun;Kim, Yong-Hoon;Hong, Gi-Hyun;Shin, Kyung-Ho;Chun, Yeon-Sook;Chun, Boe-Gwun
    • The Korean Journal of Pharmacology
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    • v.32 no.1
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    • pp.113-123
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    • 1996
  • It has been well known that polyamines ensure the stability of chromatin structure and the fidelity of DNA transcription. This study was carried out to evaluate the effect of polyamines on the apoptosis of mouse thymocytes induced by dexamethasone and polyamine synthesis inhibitors. 1) In the histological death findings of thymocytes double-stained with acridine orange and ethidium bromide, the apoptotic and the necrotic fractions (AF; NF) in the control group were $9.4{\pm}4.2%$ and $4.5{\pm}5.3%$, respectively. Dexamethasone $(3\;{\times}\;10^{-8}\;M:\;DX)$ in creased AF upto $52.0{\pm}8.1%$ and did not change NF, but A23187 $(5\;{\times}\;10^{-7}\;M:\;A2)$ increased AF and NF upto $45.0{\pm}8.9%$ and $20.5{\pm}10.6%$, respectively. 2) The thymocyte viability was significantly reduced by DX, DHEA $(1\;{\times}\;10^{-4}\;M)$, A2, DFMO $(1\;{\times}\;10^{-4}\;M)$, and $MGBG\;(1\;{\times}\;10^{-4}\;M)$, respectively. It was, however, little affected by $aminoguanidine\;(1\;{\times}\;10^{-4}\;M:\;AG)$, $putrescine\;(1\;{\times}\;10^{-5}\;M:\;PT)$, $spermidine\;(1\;{\times}\;10^{-5}\;M:\;SD)$, and $spermine\;(1\;{\times}\;10^{-5}\;M:\;SM)$. 3) The genomic DNA of mouse thymocyte was markedly fragmented by DX and A2, respectively, and to a lesser extent, by DHEA, but was little affected by MGBG, DFMO, AG, and each of polyamines. 4) The DX induced reduction of thymocyte viability was moderately attenuated by DHEA, but little affected by DFMO, MGBC, and AG. However, SM significantly attenuated the viability reduction induced by A2 as well as DX. 5) The thymocyte viability reduction by MGBG and DFMO was significantly attenuated by only SM among three polyamines applied in this study. 6) The thymocyte viability redution by combined treatments of DX with DFMO and MGBG, respectively, was significantly attenuated by SM, and moderately by PT. But the viability reduction by combined treatment of DX with AG or DHEA was not affected by polyamines. These results suggest that polyamines, particularly spermine, might play the inhibitory role in thymocyte apoptosis and the inhibitory effect can be ascribed in part to the increase of polyamine uptake by thymocytes pretreated with DFMO and MGBG.

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Epstein-Barr Virus Antibodies in Korean Mothers and Their Neonates (산모와 신생아의 Epstein Barr Virus 항체가에 관한 연구)

  • Shin, Young Kyoo;Eun, Baik Lin;Park, Sang Hee;Lim, Chae Seung;Kim, Young Sik
    • Pediatric Infection and Vaccine
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    • v.5 no.1
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    • pp.121-127
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    • 1998
  • Purposes : This study was performed to evaluate the seropositivities and levels of term pregnant women and their neonates, and the transplacental transfer rate of maternal Epstein-Barr Virus-specific IgG(VCA IgG and EBNA IgG) from term pregnant women to their neonates. Subjects and Methods : During Jan. 1, 1997 to Mar. 31. 1997, we collected the 42 pairs of sera from pregnant women and umbilical cord of their neonates in Korea University Ansan Hospital. The serum levels of VCA IgG and EBNA IgG were measured by the ELISA method. Results : 1) The seropositivities of VCA IgG were 100% in mothers and neonates. There was no statistical difference of mean VCA IgG levels between mothers and neonates. There was significant correlation of VCA IgG levels between maternal sera and neonatal umbilical cord sera(correlation coefficient r=0.5214, P<0.001). 2) The seropositivities of EBNA IgG were 100% in mothers and neonates. There was no significant difference of the mean EBNA IgG levels between mothers and neonates. There was significant correlation of EBNA IgG levels between maternal sera and neonatal umbilical cord sera (correlation coefficient r=0.7244, P<0.001). 3) There was no correlation between VCA IgG and EBNA IgG levels of maternal sera. Conclusion : Seropositivities of EBV CA IgG and EBNA IgG of term-pregnant women and their neonates were 100% and no significant differences of antibody levels were found in two groups. It seems that EBV Antibody levels in Korean mothers and neonates were high enough to protect primary EBV infection during early infancy.

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Uptake of Butachlor by Rice Seedlings and Its Phytotoxic Action to the Physiological Activities (수도묘(水稻苗)의 Butachlor 흡수(吸收) 및 약해발생(藥害發生) 특성(特性)에 관한 생리적(生理的) 연구(硏究))

  • Chung, Bong-Jin;Kwon, Yong-Woong
    • Korean Journal of Weed Science
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    • v.1 no.1
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    • pp.57-68
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    • 1981
  • To clarify the mode of uptake of butachlor (2-chloro-2', 6'-diethyl-N-(butoxymethyl) acetanilide) by rice seedlings, its phytotoxic action to growth and physiological activities, studies were conducted with rice seedlings, at the 6th or 7th leaf-stage, which were treated with nutrient solution containing butachlor 0, 1.8, 3.6, 7.2, 10.8 or 14.4 ppm for 1, 2 or 4 days, in other case, the solutions were thereafter renewed with the untreated nutrient solution for further growth. Uptake of butachlor by rice seedlings increased linearly with increase of its concentration and duration of uptake. Butachlor inhibited root growth more than shoot growth, furthermore, the inhibitory effect on the shoot growth was greater in height than in weight or leafing rate. After 4 day-treatment, the rates of shoot growth in weight were delayed for 4 days. Butachlor inhibited water uptake rapidly and linearly with increase of its external concentration. The reduced uptake of water was followed by slow increase in the stomatal resistance of leaves. Upon completion of butachlor treatment, rate of water uptake was recovered rapidly, but the stomatal resistance with lag in time. Butachlor did not affect the uptake of cation such as ammonium, potassium and calcium, but inhibited substantially uptake of nitrate in proportion to its concentration. Especially, butachlor did not affect synthesis and degradation of nitrate reductase. In addition, butachlor has shown much greater binding to the lipidic substances from rice roots than the proteinous material. The primary mechanism of phytotoxic action of butachlor does not seem to be its effect on the protein synthesis, but great affinity to membranes. The inhibition of water uptake, and its subsequent closure of stomates is thought very important for reduced growth under mild phytotoxicity.

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