• 제목/요약/키워드: in vitro cultivation

검색결과 189건 처리시간 0.023초

Isolation and Characterization of Bacteriophages Against Pseudomonas syringae pv. actinidiae Causing Bacterial Canker Disease in Kiwifruit

  • Yu, Ji-Gang;Lim, Jeong-A;Song, Yu-Rim;Heu, Sunggi;Kim, Gyoung Hee;Koh, Young Jin;Oh, Chang-Sik
    • Journal of Microbiology and Biotechnology
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    • 제26권2호
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    • pp.385-393
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    • 2016
  • Pseudomonas syringae pv. actinidiae causes bacterial canker disease in kiwifruit. Owing to the prohibition of agricultural antibiotic use in major kiwifruit-cultivating countries, alternative methods need to be developed to manage this disease. Bacteriophages are viruses that specifically infect target bacteria and have recently been reconsidered as potential biological control agents for bacterial pathogens owing to their specificity in terms of host range. In this study, we isolated bacteriophages against P. syringae pv. actinidiae from soils collected from kiwifruit orchards in Korea and selected seven bacteriophages for further characterization based on restriction enzyme digestion patterns of genomic DNA. Among the studied bacteriophages, two belong to the Myoviridae family and three belong to the Podoviridae family, based on morphology observed by transmission electron microscopy. The host range of the selected bacteriophages was confirmed using 18 strains of P. syringae pv. actinidiae, including the Psa2 and Psa3 groups, and some were also effective against other P. syringae pathovars. Lytic activity of the selected bacteriophages was sustained in vitro until 80 h, and their activity remained stable up to 50℃, at pH 11, and under UV-B light. These results indicate that the isolated bacteriophages are specific to P. syringae species and are resistant to various environmental factors, implying their potential use in control of bacterial canker disease in kiwifruits.

Tetrahymena pyriformis에 의한 마우스 복강내 대식세포의 활성화 (Effect of in vivo administration of Tetrahymena pyriformis on the in vitro toxoplasmacidal activity of mouse peritoneal macrophages)

  • 김정태;정평림;임경일
    • Parasites, Hosts and Diseases
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    • 제29권2호
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    • pp.129-138
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    • 1991
  • 자유생활 담수 섬모충류의 일종인 Tetrahymenn Pyriformis는 비특이적으로 복강 대식세포를 활성화하여 미생물 살멸효과가 있음이 밝혀졌으나 한국에서 순수분리 배양된 GL주(주)에 대한 평가는 시행된 바 없어 본 섬모충의 추출액을 ICR 마우스 복강 내로 주사하여 복강 대식세포를 활성화한 다음 실험실 내에서 표적 기생 원충인 Toxoplasma gondii (RH 주)를 접촉시켜 시간별로 Togoplasma 살멸 효과를 보아 대식세포의 환성도를 평가하였던 바 다음과 같은 결론을 얻었다. 표적 기생 원충으로서 살아있는 Toxoplasma 영양형을 주입하였을 경우, 대식세포 활성제 처리 실험군은 물론 대조군에서도 모두 처리 1분 이내에 Toxoplasma가 숙주세포내로 침입 내지 탐식되었으며 이때 탐식지수 범위는 15∼51%였다. 대조군에서는 배양 시간에 따라 100개의 대식세포당 총 탐식 표적 기생충 수가 증가하였으나 활성제 처리군에서는 배양 20시간에 그 수가 모두 줄었고 Tetrahymena 처리군에서는 총 표적 기생충 수가 14, 탐식지수 10%로서 가장 왕성한 대식세포 활성도를 나타내었다. 화학 함성 활성제로서는 dimethyldioctadecylammonium bromide (DDA)가 Tetrakymena와 대차 없는 대식세포 활성효과를 보였다. 표적 기생 편충으로서 열처리 Tonxplasma 영양형을 주입하였을 경우, 대조군을 포함한 모든 실험군에서 처리 1분 이내에 역시 Toxoplasma 가 숙주 세포에 의해 탐식되었으며 탐식지수 범위는 8∼25%여서 살아있는 영양형을 처리하였을 때보다 낮은 범위를 보였다. 이때 Tetrakymena 처리군에서는 탐식지수 4%, 대식세포 100개당 표적 세포 총수가 4로서 Texoplasma 추출액 처리군과 함께 가장 우수한 활성제로 평가되었으며 화학합성 활성제로서는 DDA, dextran sulfate, complete Freund's adjutant 순으로 환성 효과가 높았다 이상의 결과로 보아 T. pyriformis (GL 주)는 화학합성 활성제보다 우수한 Toxoplasma 살멸 효과를 나타내었고, 실험실 내에서 대량 무균 배양될 수 있어 앞으로 대식세포 활성제로서, 나아가 Toxopzasma 감염 억제제로서 널리 이용될 가능성이 있을 것으로 생각되었다.

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여름철 버뮤다그라스, 클라인그라스 및 테프그라스의 생육특성, 생산성 및 사료가치 비교 (Comparison of Growth Characteristics, Forage Production and Feed Values of Bermudagrass, Teffgrass and Kleingrass as Annual Forage Crop in Summer)

  • 박형수;최기춘;김지혜;이상훈;정종성
    • 한국초지조사료학회지
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    • 제35권1호
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    • pp.36-42
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    • 2015
  • 본 연구는 우리나라에서 여름철 조사료자원으로서 난지형목초의 재배 가능성을 검토하고 저 수분 저장 조사료 생산에 적합한 여름철 사료작물을 선발하기 위하여 난지형목초의 생육특성, 사초 생산성 및 사료가치를 평가하기 위하여 충남 천안에서 2013년 5월부터 2014년 12월까지 수행하였다. 난지형 화본과 목초인 버뮤다그라스 "Tifton 85", "Cheyenne II", "Mohawk" 3품종, 테프그라스 "Tiffany", "Emerald" 품종과 클라인그라스 "Selection 75" 품종을 매년 5월말부터 6월 초순 사이에 파종하였다. 버뮤다그라스와 클라인그라스의 출현 소요일은 파종 후 11~12일 정도 소요되었으며 테프그라스는 파종 후 2일정도 소요되는 것으로 나타났다. 버뮤다그라스는 시험기간 동안 전혀 출수가 이루어지지 않았으며, 테프그라스는 7월 10일경, 클라인그라스는 7월 24일경에 출수기에 도달하였다. 난지형 목초의 사초생산성은 버뮤다그라스 Tifton 85 품종의 건물 및 TDN 수량이 각각 13,159, 7,764 kg/ha으로 가장 높게 나타났으며 테프그라스가 가장 낮은 것으로 나타났다. NDF와 ADF 함량은 초종 간에 큰 차이가 나타나지 않았으며, 단백질 함량은 버뮤다그라스가 평균 11.5%로 난지형 목초중 가장 높게 나타났다. 총 가소화영양소(TDN) 함량은 테프그라스 Tiffany 품종이 63.6%로 가장 높게 나타났으며 in vitro 건물소화율은 초종 간에 큰 차이를 보이지 않았다. 이상의 결과를 종합해보면 우리나라에서 여름철 조사료 자원으로 난지형 목초의 재배 및 이용 가능성이 충분하였으며, 월동이 가능한 제주도와 남부지방에서는 영년생 방목초지용으로 활용성이 높고, 월동이 불가능한 중북부 지역에서는 1년생 여름철 사료작물로 이용하는 것이 우리나라에서 난지형 목초를 효율적으로 이용할 수 있는 방법으로 사료된다.

감초 육성품종 뿌리 추출물의 In vitro 활성 평가 (Evaluation of the In vitro Activity of Glycyrrhiza Cultivar Roots)

  • 이승은;이정훈;박춘근;김형돈;이윤지;서경혜;정현수;장재기;김동휘
    • 한국약용작물학회지
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    • 제27권2호
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    • pp.115-125
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    • 2019
  • Background: Glycyrrhiza radix (licorice root) have been used as an oriental medicine material for long time, and its protective effects on oxidative stress, inflammation and cognition deficit have been recently reported. However, the cultivation of Glycyrrhiza species as medicinal crops is associated with some problems such as low productivity and early leaf fall, etc. To resolve this problems, Glycyrrhiza cultivars have been developed by direct hybridization of each Glycyrrhiza species by Korean researchers. The present study was conducted to compare the Glycyrrhiza cultivar radix (Dagam, Sinwongam and Wongam) for their anti-oxidation, anti-inflammation, and cognition improvement effects and levels of liquiritin, isoliquiritigenin and licochalcone in order to select an excellent cultivar as a material resource. Methods and Results: For evaluating the inhibitory efficacies of the Glycyrrhiza cultivar extracts on oxidative stress and inflammation in BV2 cells, we measured their reactive oxygen species (ROS) production and nitric oxide (NO) release after treating them with lipopolysccharide. The scavenging activities on 2,2-diphenyl-1-picrylhydrazyl (DPPH) and peroxynitrite ($NOO^-$) radicals were evaluated. Cell proliferation and N-methyl-D-aspartate receptor (NMDAR) inhibition were analyzed. The total phenol, liquiritin, isoliquiritigenin and licochalcone A content in the extracts of the three culivars were quantified. Furthermore, the correlation coefficient between the activities and contents of total phenol, liquiritin, isoliquiritigenin and licochalcone A were also calculated. The results indicated that Sinwongam exhibited potent anti-oxidant, anti-inflammatory and NMDAR inhibititory activities. Sinwongam also showed higher total phenol and licochalcone A contents than the other cultivars. Among the three cultivars, Dagam exhibited a positive effects on NO release inhibition, cell proliferation and contents of liquiritin and isoliquiritigenin. Conclusions: Sinwongam is expected to be the most useful resource as a functional material for anti-oxidation/anti-inflammation and cognition improvement among the three studied licorice cultivars.

반지연(半枝蓮)이 HeLa Cell의 증식억제(增殖抑制)와 사멸(死滅)에 미치는 영향(影響) (Inhibitory Effects of Scutellaria barbata D. Don on the Cell Proliferation of HeLa cells)

  • 조정훈;장준복;이경섭;하지연
    • 대한한방부인과학회지
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    • 제19권4호
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    • pp.47-60
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    • 2006
  • Purpose : This study was conducted to investigate the inhibitory effects of Scutellaria barbata D. D on on the cell proliferation of HeLa Cells. Methods : Human uterine cervical carcinoma HeLa cells were cultured in the 1%, 5% and 10% concentration of Scutellaria barbata D. D on solution for 24, 48 and 72 hours for the direct inhibitory effects of Scutellaria barbata D. D on. Then we examined the effect of Scutellaria barbata D. D on solution on the cell proliferation inhibition by XTT assay. DNA fragmentation, MAP kinase activity and caspase activity by FACS analysis in HeLa cells. Results : We found that the proliferation of HeLa cells was significantly decreased in Scutellaria barbata D. D on solution containing groups comparing with a control group in a concentration-dependant manner. When HeLa cells were cultivated for 24 hours with 5% Scutellaria barbata D. D on solution containing group, the percentage of HeLa cells with activated caspase was the highest. Scutellaria barbata D. D on solution reduced the MAP kinase activity of HeLa cells comparing with the control group. By the XTT assay, the cell's activity was decreased in 5% and 10% Scutellaria barbata D. D on solution containing groups in 24 and 72 hours cultivation and 10% group in 48 hours. DNA fragmentation and caspase-3 activity of HeLa cells, however, were changed insignificantly. Conclusion : From this study we could suggest that Scutellaria barbata D. D on is available to the inhibition and apoptosis of human cervical carcinoma cell line, HeLa cells in vitro.

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$Pyoverdin_{2112}$ of Pseudomonas fluorescens 2112 Inhibits Phytophthora capsici, a Red-Pepper Blight-Causing Fungus

  • Kim, Sang-Dal;Lee, Eun-Tag;Lim, Si-Kyu;Nam, Doo-Hyun;Khang, Yong-Ho
    • Journal of Microbiology and Biotechnology
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    • 제13권3호
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    • pp.415-421
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    • 2003
  • A bacterium, Pseudomonas fluorescens 2112, that is antagonistic against a red-pepper blight-causing fungus, Phytophthora capsici, was isolated from the local soil of Gyongju, Korea. This strain formed an orange-colored clear halo zone on chrome azurol S (CAS) blue agar, suggesting the production of a siderophore in addition to an antifungal antibiotic. The optimal culture conditions for siderophore production by P. fluorescens 2112 were 30-h cultivation at $25^{\circ}C$ and pH 6.5 in King's B medium. The presence of $20{\mu}g/ml\;of\;Fe^3+$ ion or EDDHA promoted the production of siderophore in King's B medium. The siderophore was purified from culture broth by CM-Sephadex C-25 and Sephadex G-25 column chromatographies. The UV spectra of the purified siderophore was the same as that of pyoverdins or pseudobactins. The molecular mass was 1,958 Da determined by FAB-rlass spectrometer, and the amino acid composition analysis showed that the purified siderophore consisted of glycine/threonine/serine/glutamic acid/alanine/lysine with the molar ratio of 3:2:1:1:1:1, DL-Threo-${\beta}$-hydroxyaspartic acid and $N^{\delta}$-hydroxyornithine, two of the essential constituents of pyoverdin, were also found. The purified siderophore pyoverdin showed strong in vitro and in vivo antagonistic activities against phytophthora blight-causing P. capsici. Especially in an in vivo pot test, the siderophore protected red-pepper Capsicum annum L. very well from the attack of P. capsici. These results indicated that the purified siderophore of P. fluorescens 2112 played a critical role in the biocontrol of the red-pepper blight disease, equivalent to treatment by P.fluorescens 2112 cells.

Tissue-cultured regeneration and ecological values in major bamboo species

  • Sharma, Avinash;Manpoong, Chowlani;Gohain, Anwesha;Pandey, Himanshu;Padu, Gompi;Aku, Hage
    • Journal of Ecology and Environment
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    • 제46권3호
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    • pp.218-242
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    • 2022
  • Background: Promising specific growth regulators are employed in the tissue cultures of various bamboo species. Specific natural hardening mixtures support the acclimatization and adaptation of bamboo under protected cultivation. Results: The growth regulators like 2, 4-Dichlorophenoxyacetic acid (2, 4-D), Naphthaleneacetic Acid (NAA), Thidiazuron (TDZ), 6-Benzylaminopurine (BAP), Kinetin, Gelrite, Benzyl Adenine (BA), Indole Butyric Acid (IBA), Coumarin, Putrescine, Gibberellic acid (GA3), Indole Acetic Acid (IAA) has been widely used for callus induction, root regeneration and imposing plant regeneration in various species of bamboo such as Bambusa spp. and Dendrocalamus spp. Different combinations of growth regulators and phytohormones have been used for regenerating some of the major bamboo species. Natural hardening materials such as cocopeat, vermicompost, perlite, cow dung, farmyard manure, compost, soil, garden soil, and humus soil have been recommended for the acclimatization and adaptation of bamboo species. Standard combinations of growth regulators and hardening mixtures have imposed tissue culture, acclimatization, and adaptation in major bamboo species. Conclusions: Bamboo contributes to soil fertility improvement and stabilization of the environment. Bamboo species are also involved in managing the biogeochemical cycle and have immense potential for carbon sequestration and human use. This paper aims to review the various growth regulators, natural mixtures, and defined media involved in regenerating major bamboo species through in vitro propagation. In addition, the ecological benefits of safeguarding the environment are also briefly discussed.

Effect of Explant Types, Auxin Concentration and Light Condition on In Vitro Root Production and Alkaloid Content of Rauvolfia serpentina (L.) Benth. ex Kurz

  • Yahya, Andi Fadly;Hyun, Jung-Oh;Lee, Jae-Ho;Jung, Myung-Suk
    • 한국산림과학회지
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    • 제96권2호
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    • pp.178-182
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    • 2007
  • Rauvolfia serpentina (L.) Benth. ex Kurz is a medicinal plant and an endangered tropical rainforest plant species. Since the field cultivation that aims to fulfill the industrial needs is never accomplished, tissue culture appears to be the most feasible way to improve the quality and quantity of R. serpentina. This experiment used two kinds of explants (roots and shoots) to induce optimal root formation in different combinations of auxin and photoperiod. Each explants exhibited different responses on given treatments. Differentiated root could be produced from explants cultured in IBA 20 mg/L with and without light. The highest number of roots, root length and root weight induced from shoot explants were effective on MS medium containing IBA 20 mg/L and incubated under dark condition, while highest total weight (callus and root) from root explants cultured on MS medium supplemented 10 mg/L IBA and 10 mg/L NAA and incubated under day length (11/13 hr). The root induced from shoot explants produced the highest major alkaloid content. The highest content of ajmaline (2.17 ppm fresh weight) and reserpine (1.30 ppm fresh weight) were observed in shoot explants cultured in MS medium containing combination of IBA 10 mg/L and NAA 10 mg/L and incubated under dark condition, yohimbine (1.47 ppm fresh weight) was in the shoot explants cultured in MS medium containing NAA 20 mg/L and incubated under day length, while serpentine was absent.

표고버섯의 배양적 특성 및 자실체 형성에 관한 연구 (Cultural characteristics and formation of fruiting body in Lentinula edodes)

  • 이원호;김인엽;고한규;김선철;최선규;노종현;박흥수
    • 한국버섯학회지
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    • 제12권1호
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    • pp.24-28
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    • 2014
  • 표고버섯균의 균사생육에 적합한 배지로는 PDA 배지, MYA 배지로 조사되었으며 배양환경 조사로 균사생육에 적합한 온도는 $25^{\circ}C$를 나타났다. 영양원 선발 결과 탄소원은 2%의 농도에서 Malt extract를 사용하였을 때 균사생장 및 밀도가 높게 나타났다. 질소원은 0.2%의 농도에서 Yeast extract를 사용하였을 때 균사생장 및 밀도가 높게 나타났다. 표고버섯(산조701호)의 수종별 추출액 배지를 이용한 결과, 신갈나무, 굴참나무, 상수리나무, 밤나무에서는 PDA(대조구)와 같이 생장이 우수하였다. 배양기간에 따른 버섯 생산성을 조사 해본 결과, 배양 110일에서 120일에서 우수한 수확량을 보였다. 재배수종에 따른 생산성은 굴참나무에서 가장 좋은 결과를 보였다.

Laminin as a Key Extracellular Matrix for Proliferation, Differentiation, and Maturation of Porcine Muscle Stem Cell Cultivation

  • Minsu Kim;Hyun Young Jung;Beomjun Kim;Cheorun Jo
    • 한국축산식품학회지
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    • 제44권3호
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    • pp.710-722
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    • 2024
  • Extracellular matrix (ECM) proteins play a crucial role in culturing muscle stem cells (MuSCs). However, there is a lack of extensive research on how each of these proteins influences proliferation and differentiation of MuSCs from livestock animals. Therefore, we investigated the effects of various ECM coatings-collagen, fibronectin, gelatin, and laminin-on the proliferation, differentiation, and maturation of porcine MuSCs. Porcine MuSCs, isolated from 14-day-old Berkshire piglets, were cultured on ECM-coated plates, undergoing three days of proliferation followed by three days of differentiation. MuSCs on laminin showed higher proliferation rate than others (p<0.05). There was no significant difference in the mRNA expression levels of PAX7, MYF5, and MYOD among MuSCs on laminin, collagen, and fibronectin (p>0.05). During the differentiation period, MuSCs cultured on laminin exhibited a significantly higher differentiation rate, resulting in thicker myotubes compared to those on other ECMs (p<0.05). Also, MuSCs on laminin showed higher expression of mRNA related with maturated muscle fiber such as MYH1 and MYH4 corresponding to muscle fiber type IIx and muscle fiber type IIb, respectively, compared with MuSCs on other ECM coatings (p<0.05). In summary, our comparison of ECMs revealed that laminin significantly enhances MuSC proliferation and differentiation, outperforming other ECMs. Specifically, muscle fibers cultured on laminin exhibited a more mature phenotype. These findings underscore laminin's potential to advance in vitro muscle research and cultured meat production, highlighting its role in supporting rapid cell proliferation, higher differentiation rates, and the development of mature muscle fibers.