• Title/Summary/Keyword: MTT평가

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Comparative study of the biological activities effect of Mentha arvensis L. extracts from water and 80% ethanol (열수 및 80% 에탄올로 추출한 박하의 생리활성 비교연구)

  • Choi, Dong-Hun;Seung, O-Tak;Lim, Mi-Hye
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.1
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    • pp.208-216
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    • 2019
  • The purpose of this study was to investigated the biological activities such as cytotoxic, anti-oxidant and anti-inflammatory using Mentha arvensis L. extracts. Mentha arvensis L. was prepared by extracting with DW and 80% ethanol, after cell viability was assessed by MTT assay using RAW 264.7 cells. Antioxidant activities, and Anti-inflammatory activities measured through changes in the levels of reactive oxygen species (ROS), nitric oxide (NO), leukotrien B4 (LTB4), and anti- or in-flammatory cytokines($IL-1{\beta}$, IL-6, tumor necrosis factor $(TNF)-{\alpha}$, and IL-10) on LPS-induced RAW 264.7 cells. All test measured by ELISA reader and Luminex. Mentha arvensis L. was no cytotoxic in water and 80% ethanol extracts, and Concentration of 100 ug/ml of 80% ethanol extract was suppressed the productions of ROS in LPS-induced RAW 264.7 cells. Also, Productions of NO, LTB4, inflammatory or anti-inflammatory cytokines showed efficacy change that dose-independent of all extracts. In other words, Mentha arvensis L. showed significant biological activities showing anti-inflammatory, and anti-oxidant. These results may be developed as a raw material for new health food and therapeutics to ease the symptoms mentioned above.

Effect of ethanol extract of Lotus Rhizome and node of Lotus Rhizome (연근과 우절 에탄올 추출물의 향장효능 검증)

  • Jang, Young-Ah;Park, So-Hyun;Kim, Bo-Ae;Park, Jong-Yi;Jeoung, Young-Ok;Lee, Jin-Tae
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.3
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    • pp.657-665
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    • 2017
  • This study is for checking the possibility of Lotus Rhizome and node of Lotus Rhizome as cosmetic materials. For this we carried out biological active evaluation about anti-oxidant, anti-inflammatory and anti-wrinkle by using ethanol extract of Lotus Rhizome and node of Lotus Rhizome. We extracted Lotus Rhizome and node of Lotus Rhizome with 95% ethanol and then in order to evaluate anti-oxidant activity we treated samples by concentrations (100, 500, 1000) ${\mu}g/mL$ and carried out 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and The activity of 2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) cation radical scavenging. Also, in order to evaluate effect of anti-wrinkle we carried out evaluation of Elastase inhibitory activity. To evaluate effect of anti-inflammatory we evaluated toxicity of samples through MTT assay with a macrophage (Raw 264.7 cells) and measured nitric oxide production inhibitory activity. As a result, DPPH radical scavenging activity of Lotus Rhizome and node of Lotus Rhizome at $1000{\mu}g/mL$ was 66.7% and 99.5%, respectively and ABTS + radical scavenging activity was 51.2% and 98.3% at the same concentration, respectively. Elastase inhibitory activity results showed that the nodes of the Lotus Rhizom extract excellent anti-wrinkle efficacy than Lotus Rhizom. Node of Lotus Rhizome showed higher anti-wrinkle activity than the positive the control group BHT at $1000{\mu}g/mL$ concentration. According to the result of nitric oxide production inhibitory activity, Lotus Rhizom showed 55.8% effect and nodes of the Lotus Rhizom showed 66.6% effect respectively. This showed that effect of anti-inflammatory was greater in nodes of the Lotus Rhizom extracts. As a result it suggests that Lotus Rhizome and node of Lotus Rhizome extracts can be used as natural substance of cosmetics which are safe in antioxidant, anti-inflammatory, and anti-wrinkle.

In Vitro Cytotoxicity, Skin Regeneration, Anti-wrinkle, Whitening and In Vivo Skin Moisturizing Effects of Oncheongeum (온청음 물 추출물의 세포독성, 피부재생, 주름개선, 미백 및 보습 효과)

  • An, Tteul-E-Bom;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.29 no.1
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    • pp.14-34
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    • 2016
  • Objectives: The objective of this study was to evaluate the effects of cytotoxicity, skin regeneration, anti-wrinkle, whitening and skin moisturizing of Oncheongeum (OCE).Methods: The cytotoxicity of OCE lyophilized aqueous extracts (yield=13.82%) was observed against human normal fibroblast cells and B16/F10 murine melanoma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium Bromide (MTT) assay, and skin regeneration and anti-wrinkle effects were also evaluated through the assay of collagen type I synthesis compared to the transformation of the growth factor (TGF)-β1, hyaluronidase, collagenase and matrix metalloproteinase (MMP)-1 inhibitory assays compared to oleanolic acid (OA), and elastase inhibitory effects compared to phosphoramidon disodium salt (PP). In addition, OCE’s whitening effects were measured by a tyrosinase inhibitory assay and melanin formation test in B16/F10 murine melanoma cells compared to arbutin, and skin moisturizing effects were observed through a mouse skin water content test, respectively. Results: No OCE treatment-related cytotoxic effects appeared on human normal fibroblasts and B16/F10 murine melanoma cells. OCE concentration-dependently increased the collagen Type I synthesis on human normal fibroblast cells, and also effectively inhibited hyaluronidase, elastase, collagenase and MMP-1 activities. In addition, OCE inhibited melanin production of B16/F10 murine melanoma cells and activity of tyrosinase. And significant and dose-dependent increases of skin water content were detected in OCE-treated mouse skin compared to vehicle control skins. Conclusions: OCE showed favorable and sufficient effects in skin regeneration, anti-wrinkle, whitening and skin moisturizing in this experiment. But more detail mechanisms and studies on the skin protective efficiency of in vivo are needed with the screening of active biological compounds in individual OCE herbs.

The Role of NF-${\kappa}B$ in the TNF-$\alpha$-induced Apoptosis of Lung Cancer Cell Line (폐암세포주의 TNF-$\alpha$ 유발 apoptosis에서 NF-${\kappa}B$의 역할)

  • Kim, J.Y.;Lee, S.H.;HwangBo, B.;Lee, C.T.;Kim, O.H.;Han, S.K.;Shim, O.S.;Yoo, C.G.
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.2
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    • pp.166-179
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    • 2000
  • Background: The main reason for the failure of anti-cancer chemotherapy is the build up of resistance by cancer cells to apoptosis. The activation of NF-${\kappa}B$ in many cancer cell lines is reported to be underlying mechanism behind the build up of resistance of cancer cells to apoptosis. However, this relationship varied depending on the cells used in the experiments. In this study, the role of NF-${\kappa}B$ activation in the TNF-$\alpha$-induced apoptosis in lung cancer cell line was evaluated. Methods: NCI-H157 cells were used in all experiments. Cells were exposed to a high dose of TNF-$\alpha$(20 ng/ml) for 24 or 48 hours with or without blocking NF-${\kappa}B$ activation. TNF-$\alpha$-induced activation of NF-${\kappa}B$ was inhibited either by overexpression of $I{\kappa}B{\alpha}$-super repressor($I{\kappa}B{\alpha}$-SR) or by pre-treatment with proteasome inhibitor. Cell viability and apoptosis were evaluated with MTT assay and Western blot analysis for PARP fragment, respectively. Results: Cell viability of NCI-H157 cells was not affected by TNF-$\alpha$ treatment alone; however, combined treatment with TNF-$\alpha$ and cycloheximide reduced cell viability significantly, indicating that resistance to TNF-$\alpha$ is mediated by the new proteins synthesized after TNF-$\alpha$ stimulation. To evaluate the role of NF-${\kappa}B$ in the transcription of anti-apoptotic proteins. delete NF-${\kappa}B$ activation was inhibited before TNF-$\alpha$ stimulation. as described above. $AD5I{\kappa}B{\alpha}$-SR-transduction inhibited TNF-$\alpha$-induced nuclear translocation of p65. TNF-$\alpha$-induced cell death and apoptosis increased after inhibition of TNF-$\alpha$-induced activation of NF-${\kappa}$ by methods. Conclusion: These results suggest that TNF-$\alpha$-induced activation of NF-${\kappa}B$ may be closely related to the acquisition of the resistance to TNF-$\alpha$-induced apoptosis in lung cancer cells. Therefore. blocking of NF-${\kappa}B$ pathway can be a useful therapeutic modality in the treatment of lung cancer.

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Rubus coreanus Miquel Bioconversion of Metabolites by Fermenting Bacteria and Their Functional Enhancement (복분자 기능성 증대를 위한 유용미생물 활용 생물전환 공정 도입 방안 연구)

  • Lim, Jeong-Muk;Choi, Ui-Lim;Kim, Joong-Oh;Lee, Jeong-Ho;Oh, Byung-Taek
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.65-65
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    • 2018
  • 복분자(Rubus coreanus Miquel)는 anthocyanin, phenolic acid, tannin, flavonoid, stilbenoid와 같은 생리활성물질이 풍부하게 존재하는 것으로 보고되었으며 항염증, 항암활성, 항산화효과 등 다양한 생리활성에 대한 효능이 있어 건강식품으로써 애용되고 있을 뿐만 아니라 약용성 건강보조식품 등으로 각광받고 있다. 생물전환(Bioconversion)은 미생물 또는 효소의 생물학적 촉매 기반 천연소재의 생리활성 물질기능성, 생체이용률, 안전성을 증대시키기 위한 방안으로 많은 연구가 진행되고 있으며 아울러 식품, 의약품, 화장품 등 다양한 분야에서 활성화 되고 있다. 본 연구는 불가사리 발효액으로부터 유산균을 분리하였으며, 유전학적 특성을 확인하기 위하여 16S rDNA 염기서열을 분석하였다. 전북 고창에서 수확된 복분자를 분말상태로 유산균과 발효공정을 수행하였으며, 복분자의 최적 추출조건 선정과 발효공정 전 후의 활성을 관찰하였다. 발효공정 후 추출물의 기능성 평가를 진행하기 위하여 DPPH radical scavenging activity, total polyphenol 함량을 확인하여 항산화 효능 및 유효성분 함량을 평가하였다. 또한 대식세포인 Raw 264.7을 사용하여 MTT assay, Nitric oxide (NO) 생성 억제 효능을 확인하여 세포독성 및 항염증 활성을 평가하였다. 실험결과, 발효기간이 다른 불가사리 비료발효액으로부터 16 종의 다양한 균주를 확보하였으며, 생물전환 공정에 유용 균주를 선정하기 위하여 복분자 분말의 발효공정을 실시한 결과 3 종의 유산균 처리군에서 무처리군 대비 DPPH radical 소거능 및 polyphenol 함량이 증가됨을 확인하였다. 그 중 가장 우수한 활성을 나타내는 균주를 16S rDNA 염기서열 분석한 결과 Lactobacillus coryniformis A6-4로 확인되었으며, 발효공정 후 항산화 활성은 무처리군 대비 약 115%, polyphenol의 함량은 무처리군 대비 약 121%로 증가됨을 확인되었다. 또한 발효공정 후 독성활성이 감소되는 경향을 확인되었으며, 항염증 활성이 유의적으로 증가됨을 확인하였다.

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The Experimental Study on anti-inflammatory Effect of GamiJihwangTang (가미지황탕(加味地黃湯)의 항염작용(抗炎作用)에 대한 실험적(實驗的) 연구(硏究))

  • Jang, Du-Hyon;Yoo, Dong-Youl
    • The Journal of Korean Obstetrics and Gynecology
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    • v.20 no.3
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    • pp.111-128
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    • 2007
  • 목적 : 이 연구는 천식, 기관지염, 폐렴, 결핵, 산후감모 등의 호흡기 질환에 사용되는 가미지황탕(加味地黃場)의 항염작용(抗炎作用)의 효과에 대해 알아보기 위해 시행되었다. 방법 : 가미지황탕(加味地黃場)의 항염작용(抗炎作用)의 효과를 평가하기 위해 세포독성에 미치는 영향, NO, $TNF-{\alpha}$, $IL-1{\beta}$, IL-6 생성량에 미치는 영향, $TNF-{\alpha}$, $IL-1{\beta}$, IL-6 유전자 발현에 미치는 영향, iNOS, COX-2 유전자 및 단백질 발현에 미치는 영향, $PGE_2$ 합성에 미치는 영향 및 COX-2, $NF-{\kappa}B$ 활성에 미치는 영향에 대한 실험평가를 하였다. 결과 : 가미지황탕(加味地黃場)은 MTT 분석을 통한 RAW 264.7 세포주의 생존력 평가에서 세포독성이 없었고, LPS로 유도된 RAW 264.7 세포주에서 NO, $TNF-{\alpha}$, $IL-1{\beta}$ 및 IL-6 생성량을 농도 의존적으로 억제하였다. 가미지황탕(加味地黃場)은 400 g/ml 농도에서 LPS로 유도된 RAW 264.7 세포주에 대해 $TNF-{\alpha}$, $IL-1{\beta}$ 및 IL-6 유전자 발현을 농도 의존적으로 억제하였고, LPS로 유도된 RAW 264.7 세포주에서 iNOS와 COX-2 유전자 및 단백질 발현은 농도 의존적으로 억제하였다. 또한 그 농도에 따라 $PGE_2$ 생성량이 현저하게 억제하였고, LPS로 유도된 COX-2 및 $NF-{\kappa}B$ 전사활성을 농도 의존적으로 억제함으로써 iNOS와 COX-2 유전자 발현을 억제하였다. 결론 : 이상의 실험을 통해 가미지황탕(加味地黃場)은 iNOS나 COX-2와 같은 Cytokine이 있는 효소에 의해 합성되고 천식에서 증가하는, 혈관과 기관지 긴장도와 관련 있는 NO와 $PGE_2$ 생성량을 억제하고, 염증과 관련된 $TNF-{\alpha}$, $IL-1{\beta}$, IL-6의 생성량을 억제하였다. 또한 $NF-{\kappa}B$ 활성을 억제함으로써 iNOS 및 COx-2 유전자 발현을 억제하였으므로 부인과 영역에 있어서도 산후감모, 만성해수 및 천식 등의 기관지의 염증질환에 응용할 수 있을 것으로 사료된다.

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The Study of Cosmeceutical Activities from Lentinula edodes extracts and Application a Natural Cosmetic Material (표고버섯 추출물의 화장품약리활성 검증과 천연화장품 소재로써의 활용에 관한 연구)

  • Seo, Myeong-Seong;Jang, Young-Ah;Lee, Jin-Tae
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.4
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    • pp.1003-1012
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    • 2018
  • This study is for checking the possibility of Lentinula edodes as cosmetic materials. For this we carried out biological active evaluation about anti-oxidant and anti-inflammatory effects by Lentinula edodes extracts. We extracted Lentinula edodes with water and 70% ethanol and then in order to evaluate anti-oxidant activity we treated samples by concentrations (100, 500, 1000) ${\mu}g/ml$ and carried out 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, the activity of 2,2'-azino-bis ( 3-ethylbenzothiazoline-6-sulphonic acid )-diammonium salt (ABTS) cation radical scavenging and superoxide dismutase(SOD) like activity. Also, in order to evaluate effect of anti-inflammatory the samples in macrophages(RAW 264.7 cells), we carried out evaluation of cell viability, nitric oxide inhibitory activity western blot. The results of DPPH, $ABTS^+$ radical scavenging activity and SOD-like activity of the Lentinula edodes extracts increased in dose-dependent manner. The cytotoxic of samples by MTT assay showed no toxicity at the concentrations of 10, 25 and $50{\mu}g/ml$ of Lentinula edodes extract. Nitric oxide inhibition activity results showed that the extracts reduced NO productions in a concentration-dependent manner. Expression of inflammatory cytokines as $TNF-{\alpha}$, $PGE_2$ and $IL-1{\beta}$ decreased in a concentration-dependent manner and iNOS and COX-2 proteins expression rates were decreased significantly in western blot analysis. From the results of the experiment, it was comfirmed that the Lentinula edodes extracts had excellent anti-oxidant and anti-inflammatory effect and could be used as a safe natural cosmetic material in the future.

Cytotoxicity of Paraquat or Bentazone and Compensatory Effects of 3-Methylcholanthrene on the Rat Liver (Paraquat 및 Bentazone의 세포독성과 흰쥐 간에서 3-Methylcholanchrene의 독성경감효과)

  • Rim, Yo-Sup;Han, Du-Seok
    • Korean Journal of Environmental Agriculture
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    • v.20 no.3
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    • pp.155-161
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    • 2001
  • This study was carried out to investigate cytotoxicity of paraquat or bentazone on NIH 3T3 fibroblasts, toxicity of paraquat or bentazone, and compensatory effects of 3-Methylcholanthrene(3-MC) on the rat liver. In order to MTT assay, the $5.0{\times}10^4$ cell/mL of NIH 3T3 fibroblast in each well of 24 multidish were cultured. After 24 hours, the cells were treated with solution of paraquat or bentazone(1, 25, 50, 100 ${\mu}M$ respectively). After the NIH 3T3 fibroblast of all groups were cultured in same condition for 48 hours. MTT assay were performed to evaluate the cytotoxicity of cell organelles. Paraquat or bentazone $MTT_{50}$ were 1668.97 ${\mu}M$ and 1506.97 ${\mu}M$, respectively. These $IC_{50}$ of paraquat or bentazone were decided low cytotoxicity by Borenfreund. In order to observe the toxicity and compensatory effects of paraquat or bentazone on the rat liver, Sprague-Dawley male rats were used as experimental animals and divided into paraquat or bentazone only treated group and simultaneous application group of paraquat or bentazone and 3-MC. At 30 min and 1, 3, 6, 12, 24, 48 and 96 hrs interval after each treatment, the animals were sacrificed by decapitation and liver were immediately removed, immersed in fixatives, and processed with routine method for light microscopic study. Paraffin sections were stained with H-E, PAM and Best Carmine. Under the light microscope, degenerative changes of hepatic lobules were frequently observed in portal area from 3 hrs after paraquat or bentazone treatment. All hepatic cells were induced degenerative change at 12 hrs and more severe degenerative change at 48 hrs after paraquat or bentazone treatment. Especially, hepatic cells of bentazone only treated group were distinctly showed pyknotic. Glycogen granules were increased in portal area at 3 hrs, all hepatic cells at 12 hrs and remarkably increased at 48 hrs after paraquat or bentazone treated group. But hepatic cells of bentazone only treated group were regeneration at 48 hrs from portal area and glycogen granules of hepatic cells of paraquat or bentazone and 3-MC combination treated group showed in central area only at 48 hrs. The results indicate that 3-MC may be decrease paraquat or bentazone cytotoxicity on the rat liver.

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Evaluation of changes in adhesive strength and cytotoxicity of a denture adhesive according to time (시간에 따른 의치접착제의 인장 결합강도와 세포독성의 변화)

  • Jung, Ha-Yoon;Kim, Jee-Hwan;Lee, Keun-Woo;Shim, June-Sung;Moon, Hong-Seok
    • The Journal of Korean Academy of Prosthodontics
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    • v.47 no.2
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    • pp.232-239
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    • 2009
  • Statements of the problem: Many denture wearers occasionally use denture adhesives to improve denture retention, stability and chewing efficiency. An ideal denture adhesive is nontoxic, non-irritating, and provides comfort to the oral mucosa. Purpose: The purpose of this study was to evaluate the cytotoxicity and adhesive properties of a selected denture adhesive. Material and methods: To test cytotoxicity of the selected denture adhesive, mouse fibroblast cells were used in MTT testing. Cytotoxicity was examined according to the concentration of the denture adhesive and incubated for 1 to 4 days. To examine adhesive property, a denture base was fabricated on an edentulous dentiform. The adhesive was applied to the denture base, then tensile bond strength was measured, to evaluate the change in retention during 3 days. Results and Conclusion: 1. 1% and 2% concentration denture adhesive cream had no cytotoxicity. 2. The tensile bond strength of the group with both denture adhesive and artificial saliva was significantly higher than that of the group with only denture adhesive(P<.05). The tensile bond strength of the group with denture adhesive was significantly higher than that of with only artificial saliva(P<.05). 3. The tensile bond strength had no significant change during 1 hour, and then gradually decreased. After 1 day, it decrease to half. Within the limitation of this study, the tested denture adhesive had no cytotoxicilty and was effective in improving denture retention. The adhesive strength began to continuously decrease after 1 hour and it decreased to half at 1 day after application.

Evaluation of Pollen Viability of Nakdongbyeo, Two Transgenic Rice Lines, Its Hybrids with Weedy Rice, and Subsequent Selfed Progenies: F2 and F3 (낙동벼, 2개의 promoter를 각각 삽입한 유전자변형 계통과 잡초성벼(Oryza sativa)인공수정 한 후 다음세대인 F1, F2, F3의 화분활력 평가)

  • Ghimire, Sita Ram;Sohn, Eun-Young;Shin, Dong-Hyun;Lee, In-Jung;Kim, Kil-Ung
    • Journal of Life Science
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    • v.19 no.7
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    • pp.839-844
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    • 2009
  • This experiment was conducted to evaluate pollen viability of Nakdongbyeo, transgenic rice lines, an F$_1$ hybrid from a cross between Milyang weedy rice and ABC-promoter transgenic rice line containing basta-resistant (bar) gene and subsequent selfed progenies, F$_2$ and F$_3$. The reaction of pollen with 3-{4,5 dimethylthiazolyl-2}-2,5-diphenyl monotetrazolium bromide (MTT) as a staining chemical immediately after pollen shedding showed maximum pollen viability of 86% in Nakdongbeyo, 75% in ABC-promoter transgenic rice line, 62% in ubiquitin-promoter transgenic line, 68% in F$_1$, 79% in F$_2$ and 78% in F$_3$. Viability gradually declined during subsequent observations at 20-minute intervals. However, there was a drastic decline in pollen viability after 40 minutes of pollen shedding. The mean difference of pollen viability among rice lines and time was highly significant, indicating significantly different pollen viabilities at different time intervals. Maximum viability of 36.2% was observed in F$_3$ and minimum viability of 3.5% was found in F$_2$ at 90 min after pollen shedding. Results of this experiment on pollen viability and longevity elucidate potential risks of pollen-mediated flow of herbicide-resistant gene from transgenic rice lines and possible integration of it into the weedy rice population.