• Title/Summary/Keyword: petri-dish assay

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Effect of Alfalfa Plant Extracts on Germination and Early Seedling Growth of Forages

  • Chon, Sang-Uk;Choi, Seong-Kyu;Park, Sang-Won
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.45 no.5
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    • pp.310-315
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    • 2000
  • Alfalfa (Medicago sativa L.) plants have been reported to contain water-soluble substances that are autotoxic as well as allelopathic. Laboratory experiment through a petri-dish assay with imbibed seeds was conducted to evaluate both autotoxic and allelopathic effects of alfalfa leaf extracts on the germination and early seedling growth of alfalfa, red clover, crested wheatgrass, and Russian wildrye. Alfalfa seed germination was delayed dependent on extract concentration, with no difference in final germination at 72 hours. Root growth of alfalfa was stimulated up to 14% above control at very low concentrations of both leaf and stem extracts of alfalfa and was significantly reduced at extract concentration of more than 0.5g dry tissue/L (${gL}_{-1}$). Leaf extracts were generally more autotoxic for root growth than were stem extracts. Hypocotyl growth was not affected by all the concentrations of both leaf and stem extracts. Root length of legumes was more sensitive to the autotoxic chemicals from leaf extracts than was germination or shoot length. Hypocotyl growth of two legume plants and plant height of two grasses were not influenced by extracts. Seed germination and root growth of legumes were more inhibited by aqueous extracts of alfalfa leaf than were those of grasses. This result indicates autotoxic effect of alfalfa leaf extracts seems to be greater than allelopathic effect.

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Development of Seed Culture Using Soybean for Mass Production of Lovastatin with Aspergillus terreus ATCC 20542 Mutant (대두를 이용한 Lovastatin 대량생산용 Seed Culture의 제조기술)

  • Kim, Soo-Jung;Ko, Hee-Sun;Kim, Hyun-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.5
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    • pp.666-670
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    • 2008
  • Lovastatin (Mevinolin, Monacolin K) is a well-known drug for the therapy of hypercholesterolemia. It is an important fungal secondary metabolite as it inhibits 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase, EC 1.1.1.34) which catalyzes a major rate-limiting step in the biosynthesis of cholesterol. Both soybeans and black soybeans with Aspergillus terreus ATCC 20542 mutant were used as the seed culture for the mass production of lovastatin. The production of lovastatin in soybean seed culture of Asp. terreus was twofold compared to that of black soybean seed culture. The effect of two different vessels (petri dish and Erlenmeyer flask) on lovastatin production was also studied. The production of lovastatin on petri dish was tenfold to that of Erlenmeyer flask. Furthermore, the most lovastatin production on rice bran was achieved when the soybean seed culture was treated by heat shock at $30^{\circ}C$ for 1 hour, representing 82% of HMG-CoA reductase inhibition in the koji extract. We estimated that the heat treated soybean seed culture could be a new method for the mass production of lovastatin.

Evaluation of Alfalfa Autotoxicity on Germination and Early Seedling Growth of 3 Cultivars

  • Chon, Sang-Uk;Choi, Seong-Kyu
    • Plant Resources
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    • v.3 no.1
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    • pp.71-78
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    • 2000
  • Autotoxicity restricts reseeding of new alfalfa(Medicago sativa L.) after alfalfa until autotoxic chemical(s) breaks down or is dispersed into external environments, often requiring up to a year or more. One solution for reducing autotoxicity would be to select germplasms or cultivars with tolerance to the autotoxic chemical(5) and use genetically breeding program. Bioassay of seed germination and early seedling growth was conducted to evaluate autotoxic responses of 3 varieties of alfalfa to the water-soluble extracts(at 4 and 8g/L) from alfalfa ‘Cody’leaf by using agar and filter paper medium in a petri-dish assay. Root length at 5 days after seeding was more sensitive to the extract than was hypocotyl length or seed germination, and was a better parameter of autotoxic effects of alfalfa leaf extracts. Use of an agar medium gave better sensitivity of root length than did use of filter paper. Evaluating tolerance with percent of control was more important indicator than was mean of root length because of significant variation among varieties in root length of control treatment. Bioassay ranked varieties in the following order of tolerance on the basis of relative root length; “Cody” >“ Pioneer 5373” >“ Alfagraze”. Seedling growth from old “Cody” seed was more sensitive to the autotoxic chemical(5) than was that from newly produced seed.

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Wound healing effect of AHAs and moisturizing compounds with dermal equivalent (Dermal equivalent를 이용한 AHA류 및 보습 성분의 wound healing 효과에 관한 연구)

  • 유영근;정민석;최종완
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.26 no.1
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    • pp.59-80
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    • 2000
  • 본 연구는 dermal equivalent를 이용하여 AHA류(lactic acid, glycolic acid, citric acid 및 malic acid) 및 보습 원료(sodium hyaluronate, glycerine, natural carbohydrate complex)에 대한 wound healing 효과 및 그 기작을 알아보기 위한 것이다. AHA류 및 보습 원료의 독성은 dermal equivalent를 이용하여 m assay를 실시한 후 적정 농도를 petri dish위에 조성한 dermal equivalent에 처리하였다. 그리고 wound healing 효과를 알아보기 위하여 collagen 수축률을 측정하였다. 또한 collagen 합성을 촉진하는 것으로 잘알려진 vitamin C도 함께 처리하였다. AHA류의 경우 평균 pH 2-3으로 매우 낮기 때문에 4N NaOH를 이용하여 pH를 6-7사이로 조절한 sample를 함께 실험하였다. 보습 원료의 경우 전반적으로 10% 이상의 높은 NR$_{50}$보여 주었다. 특히 sodium hyaluronate(1% stock solution)의 경우에는 16% NR$_{50}$를 보여 주었다. pH를 6-7로 조절한 AHA류의 경우에는 전반적으로 0.6% 내외의 NR$_{50}$를 보이는 가운데 lactic acid는 상대적으로 높은2% NR$_{50}$를 보여 주었다. Collagen 수축률 측정 실험 결과에서는 2% sodium hyaluronate(1% stock solution)가 대조군에 비하여 처리 후 2일째 25%내외의 향상된 수축률을 보여 주었다. pH를6-7로 조절한 AHA류 중 0.1%의 malic acid의 경우에서는 대조군에 비하여 처리 후 1일째 및 2일째 각각 28% 및 35%의 수축률을 보여 주었으며 pH를 6-7로 조절한 0.1% vitamin C에서도 유사한 결과를 보여 주었다. 반면에 pH를 6-7로 조절한 0.1% citric acid의 경우에는 10-20%의 낮은 수축률을 보여 주었다. MTT assay를 이용한 UV 조사 후 pH를 6-7로 조절한 AHA류의 repairing UV damage 효과에 대한 실험에서 0.1% 및 0.01%의 malic acid와 0.01% citric acid은 irradiation control에 비하여 약 10% 이상 세포수를 증가시켰다. 그러나 예외적으로 citric acid의 경우 0.1% 농도에서 오히려 20%내외로 세포수가 감소되는 경향을 보여주었다. 그리고 lactic acid 및 glycolic acid는 두드러지는 효과를 나타내지 않았다. Collagen 합성을 측정 실험에서는 pH를 6-7로 조절한 AHA류에서는 대조군에 비하여 상대적으로 12-19% 더 합성을 촉진하였다. 반면에 pH가 2-3인 AHA류의 경우에는 대조 군과 유사하거나 조금 낮은 합성율을 보여 주었다.

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Biological Control Efficacy of an Entomopathogenic Nematode, Heterorhabditis megidis, Against Housefly, Musca domestica, and Flower Beetle, Gametis jucunda (메기디스 곤충병원선충(Heterorhabditis megidis)을 이용한 집파리와 풀색꽃무지의 생물적 방제 효과)

  • Kang Sangjin;Han Sang-Chan;Choi Kyunghee;Lee Soonwon;Kim Yonggyun
    • The Korean Journal of Soil Zoology
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    • v.8 no.1_2
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    • pp.17-22
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    • 2003
  • An endemic entomopathogenic nematode, Heterorhabditis megidis, was evaluated by its control efficacies against housefly, Musca domestica, and flower beetle, Gametis jucunda. In Petri-dish assay, the pathogenicity of H. megidis showed 456.4 infective juveniles/larva (IJs/larva) in median lethality (LC$_{50}$) against the second instar larvae of M. domestica and 238.9 IJs/larva against the second instar larvae of G. jucunda. This was contrasted with those of the other well-known entomopathogenic nematode, Steinernema carpocapsae, which showed 115.9 IJs/larva against M. domestica and 388.6 IJs/larva against G. jucunda. In field experiment, H. megidis were applied per square meter of pork farm with 1,000,000 IJs of H. megidis or apple orchard with 370,000 IJs, which were infested with M. domestica or G. jucunda, respectively. H. megidis showed 56.9% and 57.3% of control efficacies against M. domestica and G. jucunda, respectively. These results suggest a promising control technique in the field using H. megidis against M. domestica and G. jucunda.a.

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Susceptibility of the Alfalfa Weevil, Hypera postica (Coleoptera: Curculionidae) to Korean Entomopathogenic Nematodes in Laboratory Assays (한국산 곤충병원성 선충에 대한 알팔파바구미의 감수성 실내 검정)

  • Kim, Hyeong-Hwan;Han, Gun-Yeong;Park, Chung-Chan;Choo, Ho-Yul;Cho, Sung-Rae;Lee, Heung-Soo;Lee, Dong-Woon;Park, Chung-Gyoo
    • Korean journal of applied entomology
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    • v.46 no.1 s.145
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    • pp.147-151
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    • 2007
  • Laboratory tests were conducted to determine the susceptibility of the alfalfa weevil, Hypera postica to Korean entomopathogenic nematodes, Steinernema carpocapsae GSN1 strain (ScG), S. glaseri Dongrae strain (SgD), Heterorhabditis bacteriophora Hamyang strain (HbH), and Heterorhabditis sp. Gyeongsan strain (HeG) at the petri dish assay. The larval mortality of H. postica was significantly different depending on nematode species and treatment concentration. SgD and HeG strain were more effective against H. postica larva than ScG and HbH strain. When SgD and HeG strain were treated with the rate of >20 infective juveniles (ijs) per alfalfa weevil larva, mortality was ca. 77.5-100% at the late instars of H. postica in 3 days. The number of established nematode was significantly different depending on nematode species, whereas number of progeny was not significantly different. The mean number of established us of SgD strain in a host was the highest at 80 ijs by 30.2. The highest progeny number of HeG strain was 2,671.5 with 80 ijs. All nematode strains were not parasitic entirely to H. postica adults. These results show that alfalfa weevil late larva is highly susceptible to Korean entomopathogenic nematodes in the laboratory.

Effects of the root conditioning treatments after Nd:YAG laser irradiation on in vitro human gingival fibroblast attachment to root surfaces (Nd:YAG 레이저조사 후 치근의 처치방법들이 치근면 치은섬유아세포부착에 미치는 영향에 관한 연구)

  • Moon, Hye-Seong;Lim, Kee-Jung;Kim, Byung-Ock;Han, Kyung-Yoon
    • Journal of Periodontal and Implant Science
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    • v.26 no.3
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    • pp.701-713
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    • 1996
  • The purpose of this study was to evaluate the biocompatibility of the Nd:YAG lased root surface followed by root planing and/or tetracyline-HCI(T.C.-HCI) conditioning. $30,4mm{\times}4mm$ root segments were obtained from unerupted third molars and 21, periodontally involved root segments. The treatment groups were as follows : (1) healthy root cementum surface groups : 1) control(non-treated group), 2) lased only, 3) lased/root planed, and 4) lased/T.C.-HCI. (2) diseased root cementum surface groups : 1) control(root planed only), 2) lased/root planed, and 3) lased/root planed/T.C.-HCI. The specimens were treated with a Nd:YAG laser using a $320{\mu}m$ noncontact optic fiber handpiece with an energy setting of 1.5W($114.6J/cm^2$), 2.0W($152.9J/cm^2$), 5.0W($382J/cm^2$) for one minute. The fiber was held perpendicular to the petri dish(NUNC) 2cm apart in an attempt to expose the entire root segments equally. Human gingival fibroblasts were cultured from explants of normal interdental gingival tissue obtained during third morlar extraction. The attachment assay was performed with third-generation fibroblasts. The numbers of gingival fibroblasts attached to the root surface were counted on each specimen under the light microscope, and were statistically analyzed by the oneway ANOVA followed by Tukey's test in SPSS/PC+programs. The results were as follows : 1) In healthy root cementum surfaces, lased/root planed groups exhibited a significantly increased fibroblast attachment compared to controls, lased only, and lased/T.C.-HCI groups(p<0.05), 2) In diseased root cementum surfaces, laser treatment followed by root planing and/or T.C.HCl groups exhibited a increased tendency of fibroblast attachment compared to root planed only group. The results suggest that laser treatment followed by root planing and/or T.C.-HCl would appear necessary so as to render the root surface biocompatible.

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Determination of Heat Killing Temperature of Alfalfa (Medicago sativa L.) (알팔파(Medicago sativa L.)의 치사온도 결정)

  • 김기용;강경민;성병렬;김맹중;임용우;김원호;박근제;이병현
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.24 no.1
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    • pp.21-24
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    • 2004
  • To determine lethal temperature of alfalfa (Medicago sativa L. cv Vernal) at heat-stressed conditions, seedlings grown in a small pots for 4 weeks were subjected to different temperature regimes of heat treatment. No apparent demage was observed when the plants were treated at 45, 50 or $60^{\circ}C$ for 1 h. Heat treatments at 60 and $65^{\circ}C$ for 1 h, several plants were withered and showed damage symptom on their leaves. When the plants were exposed to $70^{\circ}C$ for 1 h, most of leaves were severely withered, but it was not lethal conditions for the whole plants. By contrast, most of plants were died within one day after heat treatment at $80^{\circ}C$ for 1h. Furthermore, plants exposed to $80^{\circ}C$ for 50 min were also died within 7 days. It was found that new shoots were regenerated from the plants that had been treated at $80^{\circ}C$ within 45 min. These results indicate that heat treatment at $80^{\circ}C$ for 50 min is an optimum condition to distinguish the lethality of alfalfa plants. Simple viability assay system established in this study will be useful fer selection and characterization of heat-tolerant transgenic alfalfa plants.

Determination of Heat Killing Temperature of Orchardgrass(Dactylis glomerata L.) (오차드그라스 (Dactylis glomerata L.)의 치사온도 결정)

  • Kim, K.Y.;Kang, K.M.;Rim, Y.W.;Park, G.J.;Lim, Y.C.;Seo, S.;Son, D.Y.;Jo, J.K.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.24 no.1
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    • pp.25-28
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    • 2004
  • To determine lethal temperature of orchardgrass (Dactylis glomerata L. cv. Janbeol 102) developed in Korea at heat-stressed conditions, seedlings grown in a amall pots for 4 weeks were treated at $45^{\circ}C$, $50^{\circ}C$ or $55^{\circ}C$ for 1 h. Heat treatments at $60^{\circ}C$ and $65^{\circ}C$ for 1 h, several plants were withered and showed damage symptom on their leaves. When the plants were exposed to $70^{\circ}C$ for 1 h, most of leaves were severely withered, but it was not lethal conditions for the whole plants. By contrast, most of plants were died within one day after heat treatment at $80^{\circ}C$ for 1h. Furthermore, plants exposed to $80^{\circ}C$ for 55 min were also died within 7 days. It was found that new shoots were regenerated from the plants that had been treated at $80^{\circ}C$ within 50 min. These results indicate that heat treatment at $80^{\circ}C$ for 55 min is an optimum condition to distinguish the lethality of orchardgrass plants. Simple viability assay system established in this study will be useful for selection and characterization of heat-tolerant transgenic orchardgrass plants.