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Development of a Kit for Diagnosing AtCYP78A7 Protein in Abiotic-tolerant Transgenic Rice Overexpressing AtCYP78A7 (AtCYP78A7 과발현 환경스트레스 내성 형질전환 벼의 단백질 진단 키트 개발)

  • Nam, Kyong-Hee;Park, Jung-Ho;Pack, In-Soon;Kim, Ho Bang;Kim, Chang-Gi
    • Journal of Life Science
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    • v.28 no.7
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    • pp.835-840
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    • 2018
  • Quantitative determination of the protein expression levels is one of the most important parts in assessment of the safety of foods derived from genetically modified (GM) crops. Overexpression of AtCYP78A7, a gene encoding cytochrome P450 protein, has been reported to improve tolerance to abiotic stress, such as drought and salt stress, in transgenic rice (Oryza sativa L.). In the present study, an enzyme-linked immunosorbent assay (ELISA) kit for diagnosing AtCYP78A7 protein including AtCYP78A7-specific monoclonal antibody was developed. GST-AtCYP78A7 recombinant protein was induced and purified by affinity column. Four monoclonal antibodies (mAb 6A7, mAb 4C2, mAb 11H6, and mAb 7E8) against recombinant protein were also produced and biotinylated with avidin-HRP. After pairing test using GST-AtCYP78A7 protein and lysate of rice samples, mAb 4C2 and mAb 7E8 were selected as a capture antibody and a detecting antibody, respectively, for ELISA kit. Product test using rice samples indicated that percentages of detected protein in total protein were greater than 0.1% in AtCYP78A7-overexpressing transgenic rice (Line 10B-5 and 18A-4), whereas those in negative control non-transgenic rice (Ilpum and Hwayoung) were less than 0.1%. The ELISA kit developed in this study can be useful for the rapid detection and safety assessment of transgenic rice overexpressing AtCYP78A7.

Nutritional compositions and antioxidative activities of two blueberry varieties cultivated in South Korea (국내산 두 품종 블루베리의 영양성분 및 항산화 활성 비교)

  • Song, Hyo-Nam;Park, Myoung-Su;Youn, Ho-Sik;Park, Sung-Jin;Hogstrand, Christer
    • Food Science and Preservation
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    • v.21 no.6
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    • pp.790-798
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    • 2014
  • The nutritional compositions and antioxidative activities of the Kimcheon lowbush and Pyungtaek highbush blueberries cultivated in South Korea were investigated. The approximate compositions, pH, and soluble-solid and mineral contents of P, Ca, Mg, Na, and Zn were determined. Both blueberry cultivars had mainly fructose and glucose as free sugars while disaccharides such as sucrose, maltose, and lactose were not detected in both. Oleic, linoleic, and linolenic fatty acids were the major fatty acids in both types of blueberries. The total of 16 free amino acids, including seven essential and nine non-essential amino acids, were higher in the Pyungtaek highbush blueberry. Among the amino acids, arginine was especially much higher in the Pyungtaek highbush blueberry. The anthocyanin, resveratrol, and polyphenol compounds, which are the important biologically active compounds in blueberries, were found. The anthocyanin contents of the Kimcheon lowbush and Pyungtaek highbush blueberries were 22.0 and 18.1 mg/100 g, respectively; the resveratrol contents by HPLC, 0.12 and 0.11 mg/100 g; and the total polyphenol contents, 141.3 and 138.4 mg/100 g. The electron-donating ability determined based on the DPPH radical scavenging activity was increased in a concentration-dependent manner, and it was higher than that of the Pyungtaek highbush blueberries, which implies that it is highly correlated with the higher amounts of total polyphenol, anthocyanin, and resveratrol in it. In conclusion, the two varieties of Korean blueberries can be suggested as potential sources of high-value-added functional foods.

Changes of Physicochemical Properties of Salted-Fermented Anchovy Meat Engraulis japonica with Different Salt Content During Fermentation at 15℃ (식염첨가량이 다른 멸치(Engraulis japonica)육젓의 15℃ 숙성 중 이화학적 특성의 변화)

  • LEE, Jae-Dong;KANG, Kyung-Hun;KWON, Soon-Jae;YOON, Moon-Joo;PARK, Si-Young;PARK, Jin-Hyo;KIM, Jeong-Gyun
    • Journal of Fisheries and Marine Sciences Education
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    • v.27 no.5
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    • pp.1457-1469
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    • 2015
  • This study was undertaken to investigate the quality changes of salted-fermented anchovy meat made by varying the amount of salt during fermentation at $15^{\circ}C$. Anchovy (11.0-14.0 cm of length, 10.7-17.5 g of weight) added with 15-25% of salt was filled in a round form plastic container (i.d. $10.5{\times}11cm$), and then fermented at $15^{\circ}C$ for 110 days. The factors such as proximate composition, pH, color value (L, a, b), TBA value, amino-N content, salinity, hardness value, free amino acid content and sensory evaluation of salted-fermented anchovy meat were measured. Ash content, color value (redness), TBA value, amino-N content. salinity and hardness value of salted-fermented anchovy meat were increased, but color value (lightness), and moisture content were decreased during fermentation at $15^{\circ}C$. A salted-fermented anchovy meat added with 15% of salt was shown higher content of moisture, amino-N content and free amino acid, TBA value than those of 20 or 25% of salt. Ash content, salinity and hardness value were highest in a product added with 25% of salt. From the result of sensory evaluation, Addition amount of 15% salt and fermentation periods of 110 days were determined to be the most desirable palatability of salted-fermented anchovy meat.

Effects of Harvest Date and Cultivar on the Growth, Forage Yield and Quality of Spring Sown Oats at the Middle Mountain Area (중산간지에서 수확시기와 품종이 춘파 연맥의 생육특성, 사초수량 및 사료가치에 미치는 영향)

  • Kim, W.H.;Seo, S.;Jeong, K.H.;Kim, J.G.;Shin, D.E.;Shin, J.S.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.19 no.1
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    • pp.89-94
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    • 1999
  • This experiment was carried out to determine harvest date and cultivar effects on growth characteristics, forage yield and quality of spring sown oats at the middle mountain(450m) area at the forage experimental field, Namweon Branch, National Livestock Research Institute. The experiment was arranged in a split plot design with three replication. The main plot consisted of the harvest date(9 June, 18 June). The subplots consisted of different maturities of oat cultivars such as Cayuse, Swan, Foothill, Cashel, Martlock and Winjardie. The results obtained are summarized as follows; A period of 50 days was required to be first headed from seeding with early maturity oats(Swan), but that of 77 days was required with late maturity(Foothill). The dry matter content of early maturity(Swan) oats at 9 June and 18 June were 24.01% and 35.69%, but that of late maturity cultivars(Foothill) were 14.02% and 22.84%. The fresh yield of late maturity(Foothill) oats at 9 June and 18 June were 62,666kg and 59,666kg, but that of early maturity(Cashel) were 54,222kg and 45,493kg(P<0.05). The dry yield of early maturity (Cashel) oats at 9 June was 10,169kg, but that of early maturity (Martlock) was 6,272kg. But no significant difference was found among cultivars at June 18. Crude protein content of oats were decreased from 14.0% to 11.1% as the growing stage progressed, ADF, NDF and CF contents were increased. And in vitro dry matter digestibility was decreased as the harvest date delayed. The present experiment indicated that spring sown oats(Foothill) can be successfully produced as fresh forage by seeding in middle March and harvesting in 10 and 20 June at the middle mountain (450m) area.

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Sow Transfer of Cultured Freezing Embryos by Open Pulled Straw(OPS) Methods : Preliminary Results (Open Pulled Straw(OPS) 방법에 의한 체외배양 동결수정란의 경산돈 이식 : 예비실험 결과)

  • Kim, I.-D.;Ahn, M.-H.;Hur, T.-Y.;Hong, M.-P.;Seok, H.-B.
    • Journal of Embryo Transfer
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    • v.19 no.2
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    • pp.155-163
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    • 2004
  • The aims of this study are 1) to test oocytes and embryos collected from in-vitro to achieving the valuable protocol by culturing, vitrifying and thawing of oocytes/embryos, and 2) to transfer them to recipient, and finally have resulted in pregnancies from recipient females after surgical or nonsurgical transfer. In vitro maturation and fertilization were performed according to Funahashi et al (1994). Glucose-free NCSU 23 supplemented with 5 mM sodium pyruvate, 0.5 mM sodium lactate and 4 mg/ml bovine serum albumin for 2 days at $39^{\circ}C$, and 10% fetal bovine serum albumin was added to the culture medium thereafter. Embryos were treated with 7.5 ${\mu}g/ml$ cytochalasin-B for 30 min, centrifuged at 13,000 rpm for 13 min and then exposed sequentially to an ethylene glycol(EG) vitrification solution, aspirated into OPS, and plunged/thawed into/from liquid nitrogen. In vivo embryos were surgically collected from three dornors after AI for control group. Forty-nine embryos were washed 3 times in mPBS + 10% FBS, followed treatments : cultured, centrifuged, vitrified, recovered and transferred to recipients as in vitro prepared embryos. Three recipients were transferred individually with 100, 100 frozen embryos derived from abattoir and 34 fresh embryos by surgically, and another three recipients were transferred individually with 150, 150 frozen embryos and 100 fresh embryos by nonsurgically, respectively. all recipient sows exhibited delayed returns to estrus. To our knowledge, theses results suggest that required an improved techniques, more vigorous embryos preparation and substitute to gilt with cleaner uterous condition.

Changes in Chemical and Microbiological Properties of Spring Waters in Tongyeoung Area (통영시내 약수의 화학적 및 세균학적 품질변화)

  • 최종덕;김정균
    • Journal of Food Hygiene and Safety
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    • v.15 no.4
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    • pp.328-333
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    • 2000
  • This study was carried out to changes in chemical and microbiological properties of spring waters in Tongyeoung area. In this paper, ninety spring water samples were collected from 9 station for 11 month to evaluated chemical and bacteriological water quality. Range and mean values of constituents of the samples are as followed; water temperature 5.2~25.8$^{\circ}C$, 16.3$^{\circ}C$, pH 6.0~7.2, 6.7, total residue 33.6~210 mg/1, 90.6 mg/1, turbidity 0.35~5.48, 1.45NTU, KMnO4 consumed 0.51~4.21 mg/1, 1.39 mg/1, chloride ion 6.23~42.5, 16.7 mg/l, phosphate-phosphorus ND-0.04, 0.02 mg/1, nitrite-nitrogen ND~0.02, 0.01 mg/1, nitrate-nitrogen ND~3.56, 1.42 mg/1, ammonia-nitrogen ND~0.20, 0.14 mg/1, dissolved total nitrogen ND~3.78, 1.57 mg/1, iron 0.04~0.28, 0.13ppm, zinc 0.03~0.66, 0.20ppm, mangan ND~0.01, allumium 0.14~0.58, 0.39ppm, copper ND~0.01, 0.01, lead ND~0.01, 0.01ppm, Arsenic ND~0.01, 0.01ppm, mercury ND~0.02, chrome not detected, cadmium not detetced respectively. The viable cell counts of the spring waters ranged 5.0~760/m1(means 130/m1). Range and mean value of total coliform and focal coliform MPN's of the spring waters were 0~2,400MPN/100 ml, 73MPN/100 ml and 0~540MPN/100 ml, 21MPN/100 ml. Spring water quality was usually poor with viable cell counts exceeding 130 CFU/liter and the coliform counts in spring waters of 73 MPN/liter. Composition of coliform by IMViC reaction was 33.3% E. coli, 15.6% Citrobacter freundii, 35.6% Klebsiella aerogenes and others.

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Some effects of rice weevil (Sitophilus oryzae L.) on the moisture contents of polished rice (쌀바구미(Sitophilus oryzae Linne)의 현미함수량에 미치는 영향)

  • HYUN Jai Sun
    • Korean journal of applied entomology
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    • v.1
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    • pp.20-25
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    • 1962
  • A. The increases moisture contents of grain at R.H $75\%,\; and\; 28^{\circ}C$ 1) The moisture contents of polished rice were $0.4\%,\; 1 \%,\; 3.3\%$ higher than control after 4weeks in which had been infested with 100, 200, 500 individuals of weevil at the beginning. respectively. 2) The greatest moisture contents of polished rice was $16.26\%$ for 500 individual infested grain and the lowest one was $14.86\%$ for the 100 individual weevil infested grain after 4 weeks. 3) The increases in moisture contents of grain were proportionally greater with the size of the weevil population. B. Changes in moisture contents of polished rice at the three levels of cylinder which contain 3kgs of grain and 1000 weevils were infested at the beginning of the experiment. 1) The moisture contents of the grain at all the levels of the cylinder which was not infested with weevil, were consistantly decreased in 12 weeks. 2) The moisture contents of the grain at all the levels of the cylenders which had been infested with 1000 weevils at the beginning of the experiment, were increased except top lovel, that were $15.6\%$ for middle, $41.55\%$ for the bottom levels after 12 weeks. 3) The moisture contents of the grain were greater toward bottom in both weevil infested grain and in control. 4) The moisture content increase might be caused by the respiration of the weevils and deterioration of the grain by microorganisms. 5) There were no tendency to confine the weevils at any definite levels, but as the deterioration of the grain goes on, the weevil moved to top levels of cylinders. 5) The decreases in moisture contents of the grain at the top levels of the cylinders might be caused by diffusion of water to the environment. 7) The differences in temperature at three levels of the cylinder from enviroment were little or negative in control but were greater and positive. and the bottom were higher than top in experimental cylinders. 8) The increases of temperature might be caused by the respiration of the weevil and microorganisms.

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Symbiotic effectiveness and intrinsic antibiotic resistance of Rhizobium meliloti populated in Korean pasture soils (국내(國內) 초지토양(草地土壤)에 분포(分布)한 Rhizobium meliloti의 질소고정력(窒素固定力)과 항균제반응(抗菌劑反應) 특성(特性))

  • Kang, Ui-Gum;Ha, Ho-Sung;Jung, Yeun-Tae
    • Applied Biological Chemistry
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    • v.35 no.3
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    • pp.179-185
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    • 1992
  • Rhizobium meliloti populated in five Korean pasture soils were characterized by symbiotic effectiveness and intrinsic antibiotic resistance using whole-soil inoculum and 11 antibiotics, respectively. Most probable number (MPN) of naturalized rhizobia counted with alfalfa Vernal[Medicago sativa (L.)] as a host ranged $1.7{\times}10^2\;cells/g$. soil(Chunghyo, Kyeongiu)-$1.0{\times}10^5\;cells/g$. soil(Gampo, Kyeongiu) and ended to be positively associated with soil pH. On the whole, the effectiveness of population as compared to TAL mix inoculum (TAL 380+TAL 1372+TAL 1373) was very low. Nevertheless, there were two highly effective strains, YCK 539 and YCK 542, which were not inferior to TAL 1372, from Ogpo, Dalseong among the total of 30 of 6 isolates per each soil. As long as mean $N_2$ fixing ability of each soil isolate, the isolates from Hyeongog, Kyeonju were outstanding and the rest were in order of Ogpo, Dalseong>Chunghyo, Kyeongju>Hwaweon, Dalseong>Gampo, Kyeongiu. Isolates as a whole were resistant to erythromycin(67崙), nalidixic acid(77%), and streptomycin sulfate(8051), which had the concentration of $100\;{\mu}g/ml$, $160\;{\mu}g/ml$, and $10\;{\mu}g/ml$, respectively and divided into 14 patterns of resistance. Association between resistances in each soil was not clear. And there was no relationship of resistance pattern to effectiveness. The best effective strain YCKa 542 exclusively fell into No. X pattern having resistance to erythromycin, nalidixic acid, and neomycin sulfate.

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Biological Activities and Bioactive Compounds in the Extract of Acer tegmentosum Maxim. Stem (산겨릅나무 줄기추출물의 생리활성 및 유효성분 분리)

  • Hong, Bo-Kyong;Eom, Seok-Hyun;Lee, Chan-Ok;Lee, Ji-Won;Jeong, Jong-Hyun;Kim, Jae-Kwang;Cho, Dong-Ha;Yu, Chang-Yeon;Kwon, Yong-Soo;Kim, Myong-Jo
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.4
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    • pp.296-303
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    • 2007
  • Acer tegmentosum (Acereaceae) has been used a source of traditional medicines for the treatment of hepatic disorders in Korea. This research was conducted to determine biofunctional activities of A. tegmentosum stem extract and to identify its bioactive components. Methanolic extract from A. tegmentosum stem was partitioned by using organic solvents, including n-hexane, ethyl acetate, n-butanol, and water. Two compounds were isolated by using an ODS column chromatography from ethyl acetate soluble fraction shown to the strongest antioxidant activity ($RC_{50}=3.15\;{\mu}g/m{\ell}$) among the fractions. The isolated compounds were analyzed by $^1H$ and $^{13}C$ NMR, IR, UV/VIS, MS spectrum data and identified as catechin, ${\rho}-Hydroxyphenethyl$ alcohol $1-O-{\beta}-_D-(6'-O-galloyl)-glucopyranoside$. The compounds have shown strong antioxidant activity, with similar activity to BHA ($RC_{50}=2\;{\mu}g/m{\ell}$). Especially, ${\rho}-Hydroxyphenethyl$ alcohol 1-O-{\beta}-_D-(6'-O-galloyl)-glucopyranoside$ was shown strong anti-lipid peroxidative activity. However, the compounds were not shown antimicrobial activities. In antimicrobial activity assays, ethyl acetate soluble fraction was effective to bacterial inhibition, such as Escherichia coli and Klebsiella pneumonia, with minimum inhibitory concentrations in $125\;{\mu}g/m{\ell}$. Otherwise, antifungal activity against Candida albicans was shown in n-hexane soluble fraction exhibiting $63\;{\mu}g/m{\ell}$ of minimum inhibitory concentration. In anticomplementary activity assays, water soluble fraction was the most effective exhibiting 24% inhibitory activity.

The Responses of Antioxidative Enzymes and Salt Tolerance of Atriplex gmelini (Atriplex gmelini(가는갯능쟁이)의 내염성과 항산화 효소 반응)

  • 배정진;윤호성;추연식;송승달
    • The Korean Journal of Ecology
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    • v.26 no.5
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    • pp.273-280
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    • 2003
  • Saline conditions invoke oxidative stress attributed to the overproduction of reactive oxygen species (ROS). Changes in quantum efficiency and antioxidative enzyme activity upon salt treatment were examined in a salt-tolerant plant, Atriplex gmelini, to test the hypothesis that salt tolerance of A. gmelini is due to the increased activity of antioxidative enzymes. A. gmelini showed optimum growth at 100 mM NaCl producing 116% of the shoot dry weight over control plants in 0 mM NaCl treatment. Healthy growth persisted up to 300 mM NaCl treatment maintaining normal internal water content and dry weight. No photochemical stress or damages on antioxidative defense system was obvious in plants of 2 and 4 day salt treatment which was indicated by increased quantum efficiency (Fv/Fm value), decreased stress index (Fo/Fm value), and increased activity of antioxidative enzymes such as SOD, APX, GR. However, the plants treated with 400 mM NaCl showed decrease in growth and in antioxidative enzyme activity although the enzyme activity was still higher than that of the 0 mM NaCl treated plants (l31%, 114%, and 134% of the SOD, APX, and GR activity, respectively). Interestingly, another important antioridative enzyme that scavenges H₂O₂ in plant cells, CAT, showed rapid decrease in its activity as salt concentration increased; 38%, 22%, 15% of the 0 mM NaCl treated plants at 200, 300, 400 mM NaCl treatments, respectively. It appears that the enzymes in ascorbate-glutathione cycle such as APX and GR play the major roles in scavenging ROS produced by salt stress in A. gmelini. After 6 days of salt treatment, the damage in photochemical and antioxidative defense system was indicated by decreased Fv/Fm value and increased Fo/Fm value. A. gmelini appears to cope with short term salt treatment by enhanced activity of the antioxidative defense system, whereas long term stress invoke oxidative stress by increased ROS due to the damages in photochemical and antioxidative system.