• Title/Summary/Keyword: plant cells

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Molecular Cloning and Function Analysis of an Anthocyanidin Synthase Gene from Ginkgo biloba, and Its Expression in Abiotic Stress Responses

  • Xu, Feng;Cheng, Hua;Cai, Rong;Li, Lin Ling;Chang, Jie;Zhu, Jun;Zhang, Feng Xia;Chen, Liu Ji;Wang, Yan;Cheng, Shu Han;Cheng, Shui Yuan
    • Molecules and Cells
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    • v.26 no.6
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    • pp.536-547
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    • 2008
  • Anthocyanidin synthase (ANS, leucoanthocyanidin oxygenase), a 2-oxoglutarate iron-dependent oxygenase, catalyzed the penultimate step in the biosynthesis of the anthocyanin class of flavonoids, from the colorless leucoanthocyanidins to the colored anthocyanidins. The full-length cDNA and genomic DNA sequences of ANS gene (designated as GbANS) were isolated from Ginkgo biloba for the first time. The full-length cDNA of GbANS contained a 1062-bp open reading frame (ORF) encoding a 354-amino-acid protein. The genomic DNA analysis showed that GbANS gene had three exons and two introns. The deduced GbANS protein showed high identities to other plant ANSs. The conserved amino acids (H-X-D) ligating ferrous iron and residues (R-X-S) participating in 2-oxoglutarate binding were found in GbANS at the similar positions like other ANSs. Southern blot analysis indicated that GbANS belonged to a multi-gene family. The expression analysis by real-time PCR showed that GbANS expressed in a tissue-specific manner in G. biloba. GbANS was also found to be up-regulated by all of the six tested abiotic stresses, UV-B, abscisic acid, sucrose, salicylic acid, cold and ethylene, consistent with the promoter region analysis of GbANS. The recombinant protein was successfully expressed in E. coli strain with pET-28a vector. The in vitro enzyme activity assay by HPLC indicated that recombinant GbANS protein could catalyze the formation the cyanidin from leucocyanidin and conversion of dihydroquercetin to quercetin, suggesting GbANS is a bifunctional enzyme within the anthocyanidin and flavonol biosynthetic pathway.

Expression of the Floral Repressor miRNA156 is Positively Regulated by the AGAMOUS-like Proteins AGL15 and AGL18

  • Serivichyaswat, Phanu;Ryu, Hak-Seung;Kim, Wanhui;Kim, Soonkap;Chung, Kyung Sook;Kim, Jae Joon;Ahn, Ji Hoon
    • Molecules and Cells
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    • v.38 no.3
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    • pp.259-266
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    • 2015
  • The regulation of flowering time has crucial implications for plant fitness. MicroRNA156 (miR156) represses the floral transition in Arabidopsis thaliana, but the mechanisms regulating its transcription remain unclear. Here, we show that two AGAMOUS-like proteins, AGL15 and AGL18, act as positive regulators of the expression of MIR156. Small RNA northern blot analysis revealed a significant decrease in the levels of mature miR156 in agl15 agl18 double mutants, but not in the single mutants, suggesting that AGL15 and AGL18 co-regulate miR156 expression. Histochemical analysis further indicated that the double mutants showed a reduction in MIR156 promoter strength. The double mutants also showed reduced abundance of pri-miR156a and pri-miR156c, two of the primary transcripts from MIR156 genes. Electrophoretic mobility shift assays demonstrated that AGL15 directly associated with the CArG motifs in the MIR156a/c promoters. AGL18 did not show binding affinity to the CArG motifs, but pull-down and yeast two-hybrid assays showed that AGL18 forms a heterodimer with AGL15. GFP reporter assays and bimolecular fluorescence complementation (BiFC) showed that AGL15 and AGL18 co-localize in the nucleus and confirmed their in vivo interaction. Overexpression of miR156 did not affect the levels of AGL15 and AGL18 transcripts. Taking these data together, we present a model for the transcriptional regulation of MIR156. In this model, AGL15 and AGL18 may form a complex along with other proteins, and bind to the CArG motifs of the promoters of MIR156 to activate the MIR156 expression.

Effects of polysaccharide fractions from phellodendron chinese SCHNEID on tumor progression and immunopontentiation

  • Jun, Kya-I;Lee, Tae-Kyun;Kim, Cheorl-Ho
    • Advances in Traditional Medicine
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    • v.1 no.1
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    • pp.37-44
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    • 2000
  • In the previous paper (Kim et al., Glycoconjugate Journal 16, 247-252, 1999), heteropolysaccharides from Korean medicinal plant, Phellodendri cortex (Hwangbek) showed a poten B-Iymphocyte-stimulating activity in a system using polyclonal antibody forming cells in C57BL/6XC3H mice at dosages of 2-10 mg. In a series of biolgical active polysaccharides from natural medicinal plants, the polysaccharide fractions were isolated and purified from Phellodendron chinese SCHNEID, and antitumor activities were examined at dosages of 2, 5 and 10 mg/100 g. F-7 and F-8 showed the highest tumor inhibitory activities (inhibition ratio 96.4 and 98.2% in 2 mg/100 g), and in dose of 5 mg/100 g, the inhibitory ratios were 95.3 and 97.5%, respectively. Furthermore, 10 mg/100 g of intraperitoneal (i.p.) injection gave 97.3 and 98.7% of inhibition. In oral administration, the inhibitory activities were not markedly observed, indicating that the polysaccharides are directly acting to immune system. When the effects on TS and TK activities were determined, TS activities in the F-2 and F-7-treated mice were markedly suppressed to 73.7% and 79.5% of that in the control (p<0.01), while there was little difference in TK activity with a slight decrease in F-2 only. However, in i.p. injection, TS activities in the F-2, F-5, F-7 and F-8-treated mice were markedly suppressed to 83% to 85% of that in the control (p<0.01). Furthermore, there was also significant differences in TK activities in F-2, F-5, F-7 and F-8-treated mice (p<0.05). Therefore, polysaccharide fraction F-8 was further purified to active fractions of F-9 and F-11 by gel permeation chromatography using TSK Gel HW50S. The purified polysaccharides of F-9 and F-11 were composed of GlcNAc (47.3%), Gal (24.7%) and Man (28.0%). These results clearly indicated that the i.p. injection is much effective to suppress tumor growth than oral administration.

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Development of High Performance Nanocomposites using Functionalized Plant Oil Resins (식물성오일 레진을 이용한 고기능성 나노 복합재료의 개발)

  • Han, Song-Yi;Jung, Young-Hee;Oh, Jeong-Seok;Kaang, Shin-Young;Hong, Chang-Kook
    • Elastomers and Composites
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    • v.47 no.1
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    • pp.2-8
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    • 2012
  • In this study, in order to develop renewable bio-based nanocomposites, multi-functional nanocomposites from soybean resins (AESO, MAESO) and nanoclay were prepared. Photoelectrodes for environmental friendly dye-sensitized solar cell using soybean resin were also prepared. Organo-modified nanoclay was directly dispersed in functionalized soybean resins after mixing with styrene as a comonomer and radical initiator was used to copolymerize the nanocomposites. The observed morphology was a mixture of intercalated/exfoliated structure and the physical properties were improved by adding nanoclay. A nanocomposite using MAESO, which added COOH functional group to the soybean resin, showed better dispersibility than AESO composites. Ultrasonic treatment of the nanocomposites also improved the physical properties. Nanoporous $TiO_2$ photoelectrode was also prepared using soybean resins as a binder, after acid-treatment of $TiO_2$ surface using nitric acid. Dye-sensitized solar cells were prepared after adsorbing dye molecules on it. The $TiO_2$ photoelectrode prepared using soybean binder had high current density because of increased surface area by improved dispersibility. The photoelectrochemical properties and conversion efficiency of the solar cell were significantly improved using the soybean binder.

Anti-inflammatory and Anti-bacterial Active Ingredients Derived from the Extract of the Leaves of Hydrangea Petiolaris (등수국 잎 추출물 유래 항염 및 항균 활성 성분)

  • Jo, Seong Mi;Kim, Jung Eun;Lee, Nam Ho
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.3
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    • pp.207-218
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    • 2020
  • In this study, the anti-inflammatory and anti-bacterial activities of the extracts from the leaves of the Hydrangea petiolaris were identified, and the chemical structure was identified by separating the active ingredient. As the result of the anti-inflammatory activity experiment using RAW 264.7 cells, it was confirmed that the n-hexane (Hex) and ethyl acetate (EtOAc) fractions inhibited the production of nitric oxide (NO) and the expression of iNOS protein in a concentration-dependent manner without cytotoxicity. In addition, the n-Hex and EtOAc fractions reduced the production of pro-inflammatory cytokines (TNF-α, IL-1β and IL-6). Upon the anti-bacterial tests using Staphylococcus epidermidis and Cutibacterium acnes, the extract, n-Hex, EtOAc and n-butanol (BuOH) fractions showed potent activities. In order to isolate the active constituents, the n-Hex and EtOAc fractions were further purified to afford four phytochemicals; phytol (1), corosolic acid (2), asiatic acid (3) and 1-O-p-coumaroyl-β-D-glucopyranoside (4). All of the compounds 1 - 4 were isolated for the first time from this plant. In addition, the contents of isolated compounds were determined by HPLC and the quantity of phytol (1) was 27.8 mg/g for the 70% EtOH extract. Based on the above research results, it is believed that it will be possible to develop a natural cosmetic material that has anti-inflammatory and anti-bacterial effects using the extract of H. petiolaris leaves.

Antioxidant Activity and Inhibitory Effect of Taraxacum officinale Extracts on Nitric Oxide Production (서양민들레 부위별 추출물의 항산화활성 및 nitric oxide 생성저해효과)

  • Min, Kyung-Chun;Jhoo, Jin-Woo
    • Korean Journal of Food Science and Technology
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    • v.45 no.2
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    • pp.206-212
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    • 2013
  • In the current study, antioxidant properties of extracts from different parts of Taraxacum officinale were determined by measuring the radical scavenging activity of 1,1-diphenyl-2-picryl hydrazyl(DPPH) and 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). The antioxidant activity of aerial and root parts of this plant was measured in extracts obtained from various solvents, e.g., water, 50, 70, 100% ethanol, and hot-water. The hot-water extracts had the highest radical scavenging activities. A further study was conducted to determine the total antioxidant activity of hot-water extract fractions obtained from different solvents, e.g., chloroform, ethyl acetate, n-butanol, and water. The ethyl acetate fraction of this extract displayed the strongest DPPH and ABTS radical scavenging activity and the highest total phenolic contents. The inhibitory effect of individual solvent fractions on the production of nitric oxide in LPS-stimulated RAW 264.7 cells was measured. The results indicated that the ethyl acetate fraction of aerial and roots part extracts significantly reduced nitric oxide productions.

Skin Whitening Effect of Sedum kamtschaticum Fisch. Solvent Fractions (기린초(Sedum kamtschaticum Fisch.) 추출 분획물의 피부 미백효과 연구)

  • Yoon, Jihye;Park, Jihye;Kim, Bora
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.5
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    • pp.1239-1247
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    • 2020
  • Sedum kamtschaticum Fisch., a native plant of Korea, has been used in Korean traditional medicine in the form of water extract for its capacity to improve blood circulation and for its antioxidant and anti-inflammatory effects. Since previous research suggests that S. kamtschaticum Fisch. has excellent antioxidant and mushroom tyrosinase inhibition activities, in this study, the root and stem parts of S. kamtschaticum Fisch. are extracted in 70% ethanol (SKS, SKR), fractionated with and in order of n-hexane (SSH), ethyl acetate (SSE, SRE), chloroform (SSC, SRC) and water (SSW, SRW) according to the polarity of each solvent, and tested for its applicability as a cosmetic material. According to the total polyphenol, flavonoid contents and DPPH radical scavenging activity of each fraction, the contents and scavenging activity of the root extractions (SKR) were higher than those of the stem extractions (SKS), ethyl acetate fractions (SSE, SRE) being the most effective. In addition, ethyl acetate fractions had the highest tyrosinase inhibition activity and melanin synthesis inhibition activity used on B16F10 melanoma cells, at the concentration of 10 ㎍/mL. HPLC analysis detected a variety of polyphenols including gallic acid and quercetin. This study suggests the potential role of S. kamtschaticum Fisch. as a natural cosmeceutical material.

Comparison of Algal Growth Kinetics using Reclaimed Wastewaters from Various Treatment Processes (다양한 수질정화 공정 별 하수처리수 재이용수의 조류성장 비교)

  • Joo, Jin-Chul;Seo, Sou-Hyun;Song, Ho-Myeon;Kim, Il-Ho;Ahn, Chang-Hyuk
    • Proceedings of the Korea Water Resources Association Conference
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    • 2011.05a
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    • pp.309-309
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    • 2011
  • 지구온난화와 도시기후 변화에 대응하기 위해 자연의 생태적 기능을 복원하고 환경에 대한 오염부하를 저감하여 도시 환경의 건강성과 지속성을 높이기 위해 도심 내 물순환시스템(urban water circulating system)의 구축이 요구된다. 즉, 물순환시스템을 활용하여 도심 내 다양한 수원(생태하천/호수 유지용수, 하수처리수, 우수, 지하수 등)을 네트워크 및 통합 관리하여 도시 내 물순환의 건전성과 수자원의 재이용률을 향상시킬 수 있다. 이를 위해서 연중 발생량이 일정하고 막대한(66.4억톤/년, 2009년 기준) 하수처리수 방류수는 고도처리를 통해 수질이 양호하며 안정적인 대체 수자원으로 고려된다. 또한, 하수처리수의 재이용은 공공수역으로 배출되는 오염부하량의 총량 삭감 및 상수사용량의 절감과 수자원을 효율적으로 이용한다는 면에서 최근 재이용 사례가 증가하고 있는 추세이다. 그러나, 도심 내 친수공간(생태하천/호수)은 저류수량에 비해 유입수량이 적어 체류시간이 비교적 장시간이고, 이로 인해 부영양화가 쉽게 발생해 수질이 악화된다. 따라서, 본 연구에서는 하수처리수 재이용수를 도심 내 친수공간의 유지용수로 활용 시, 수질정화공정(응집 후 여과, 응집 후 여과+한외여과, 응집 후 여과+한외여과+역삼투 공정)이 친수공간 내 조류성장에 미치는 영향을 파악하기 위해, 하수처리수 재이용수 pilot plant의 수질정화공정별 유출수를 활용해 M. aeruginosa를 시험조류로 조류성장(growth kinetics)을 조사하였다. 조류는 $5\times104$ cells/mL의 초기 농도로 접종하여 배양하였으며, 조류성장에 직접적인 제한인자인 용존반응성인의 농도에 따른 성장속도를 Monod와 변형 Monod Kinetics를 이용해 반포화상수(Ks)와 최대 성장속도(${\mu}$max)를 산정하였다. 실험결과, 역삼투 공정을 제외한 다른 수질정화공정은 비록 영양염류가 80~90% 이상 제거되어 수계의 화학적 성상이 변하였으나 조류성장역학의 변화는 통계학적 (p=0.05)으로 유의할만한 수준은 아닌 것으로 판명되었다. 또한, 수리학적 체류시간이 2주 이상이 될 경우, 역삼투 공정을 제외한 수질정화공정 별 유출수에서는 조류의 과다성장으로 인해 부영양화가 발생하는 것으로 판명되었다. 결론적으로 하수처리수 재이용수를 친수용수로 활용시, 조류성장을 방지하기 위하여 용존반응성인의 농도를 중점적으로 관리하는 수질정화공정 및 유지용수 공급방안을 고려해야하는 것으로 판단된다.

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Effect of ZnO and TiO2 Nanopaticles (NPs) on Microorganisms Growth in Activated Sludge (활성슬러지에서의 미생물 성장에 대한 ZnO와 TiO2 나노물질의 영향)

  • Ha, Min Jeong;Lee, Yeo Eun;Jang, Am
    • Journal of Korean Society of Environmental Engineers
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    • v.38 no.4
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    • pp.177-183
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    • 2016
  • Sewage treatment using microorganisms is affected by multiple factors such as microbial properties, characteristics of sewage and operating conditions, and nanoparticles inflow may cause negative effects on sewage treatment system especially on the system stability and efficiency. It was studied to assess the toxic effects of nanoparticles on microorganism growth. The activated sludge in the sewage treatment plant of university was cultured in the optimized medium for each strain. Bacillus (gram-positive), Pseudomonas and E.coli (gram-negative) in the activated sludge were selected as target microorganisms, and ZnO and $TiO_2$ were chosen as nanoparticles. For same concentration of nanoparticles, average growth inhibition rate of Bacillus was 60% or more, while that of Pseudomonas was less than 10%. The toxicity of nanoparticles was shown to be higher for gram-positive bacteria than gram-negative bacteria because of their differences on structure of cell wall, components of cell wall protein, physiology of cells and metabolism. ZnO affected 3 times more negative on the growth of microorganisms as compared to $TiO_2$. It was assumed that, therefore, toxicity of ZnO was found to be greater than $TiO_2$.

Microbial Communities of the Microbial Fuel Cell Using Swine Wastewater in the Enrichment Step with the Lapse of Time (가축분뇨를 이용한 미생물연료전지의 농화배양 단계에서 미생물 군집 변화)

  • Jang, Jae Kyung;Hong, Sun Hwa;Ryou, Youg Sun;Lee, Eun Young;Chang, In Seop;Kang, Young Koo;Kim, Jong Goo
    • Journal of Korean Society of Environmental Engineers
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    • v.35 no.12
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    • pp.973-977
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    • 2013
  • These studies were attempted to investigate the change of microbial community of anode of microbial fuel cell using swine wastewater in the enrichment step with the lapse of time. Microbial fuel cells enriched by a 1 : 1 mixture of anaerobic digestive juices of the sewage treatment plant and livestock wastewater. Enrichment culture step was divided into three stages to indentify the microorganisms. It was separated by each lag phase, exponential phase, and stationary phase. These steps were determined by the change of the current value. The current after enrichment was generated about $0.84{\pm}0.06mA$. We were cut out the different 17 bands in the DGGE fingerprint gel to do sequencing. The bands which the concentration was increasing or newly appearing with the lapse of time were included for this study. In the lag and exponential phase, Clostridium, Rhodocyclaceae, Bacteriodetes, and Uncultured bacterium etc. were detected. There were in the stationary phase Geobacter sp., Rhodocyclaceae, Candidatus, Nitrospira, Flavobactriaceae and uncultured bacterium etc. Geobactor among microorganisms detected in this study is known as the Electrochemically active microorganisms. It may include electrochemically active microorganisms to be considered as electrical activity microorganisms.