• Title/Summary/Keyword: Plant Cells

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Isolation of Circadian-associated Genes in Brassica rapa by Comparative Genomics with Arabidopsis thaliana

  • Kim, Jin A;Yang, Tae-Jin;Kim, Jung Sun;Park, Jee Young;Kwon, Soo-Jin;Lim, Myung-Ho;Jin, Mina;Lee, Sang Choon;Lee, Soo In;Choi, Beom-Soon;Um, Sang-Hee;Kim, Ho-Il;Chun, Changhoo;Park, Beom-Seok
    • Molecules and Cells
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    • v.23 no.2
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    • pp.145-153
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    • 2007
  • Elucidation of the roles of circadian associated factors requires a better understanding of the molecular mechanisms of circadian rhythms, control of flowering time through photoperiodic pathways, and photosensory signal transduction. In Arabidopsis, the APRR1 quintet, APRRs 1, 3, 5, 7, and 9, are known as central oscillator genes. Other plants may share the molecular mechanism underlying the circadian rhythm. To identify and characterize these circadian response genes in Brassica crops whose genome was triplicated after divergence from Arabidopsis, we identified B. rapa BAC clones containing these genes by BLAST analysis of B. rapa BAC end sequences against the five corresponding Arabidopsis regions. Subsequent fingerprinting, Southern hybridization, and PCR allowed identification of five BAC clones, one for each of the five circadian-related genes. By draft shotgun sequencing of the BAC clones, we identified the complete gene sequences and cloned the five expressed B. rapa circadian-associated gene members, BrPRRs 1, 3, 5, 7, and 9. Phylogenetic analysis revealed that each BrPRR was orthologous to the corresponding APRR at the sequence level. Northern hybridization revealed that the five genes were transcribed at distinct points in the 24 hour period, and Southern hybridization revealed that they are present in 2, 1, 2, 2, and 1 copies, respectively in the B. rapa genome, which was triplicated and then diploidized during the last 15 million years.

Isolation and quantitative analysis of metabolites from Scrophularia buergeriana and their hepatoprotective effects against HepG2 Cells (현삼 (Scrophularia buergeriana)에서 분리한 화합물의 함량분석 및 간세포 보호 효과)

  • Na, Hyeon Seon;Oh, Seon Min;Shin, Woo Cheol;Bo, Jeon Hwang;Kim, Hyoung-Geun;Yoon, Dahye;Yang, Seung Hwan;Lee, Young-Seob;Kim, Geum-Soog;Baek, Nam-In;Lee, Moon-Soon;Lee, Dae Young
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.399-406
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    • 2019
  • The roots of Scrophularia buergeriana were extracted with 80% aqueous Methanol and the concentrates were partitioned into EtOAc, n-BuOH, and H2O fractions. The repeated silica gel or octadecyl SiO2column, and medium pressure liquid chromatographies for the n-BuOH fraction led to isolation of phenylethanoid glycosides and iridoid glycosides. The chemical structures of these compounds were determined as harpagoside (1), angoroside C (2), aucubin (3) and acetoside (4) based on spectroscopic analyses including nuclear magnetic resonance and MS. A simple and efficient HPLC with UV detection method for the simultaneous determination of the four compounds (1-4) has been developed and applied to their content determination in the S. buergeriana. The roots were extracted by 80% methanol, and the contents of 1, 2, 3, and 4 were determined to 11.5, 7.6, 41.2, and 4.8 mg/g, respectively. Additionally, angoroside C (2) and acetoside (4) exhibited hepatoprotective effect against ethanol-induced hepatotoxicity in HepG2 cell line.

Chemical Suppression of Gravitropic Bending Response in Flower Stalks of Snapdragon (Antirrhinum majus L.) (몇가지 생리활성 저해제가 금어초 절화의 굴지성 반응에 미치는 효과)

  • Kim, Yong-Sam;Kim, Dong-Hern;Hwang, Young-Soo;Jung, Jin
    • Applied Biological Chemistry
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    • v.40 no.6
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    • pp.567-571
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    • 1997
  • Numbers of chemical agents which have been shown to inhibit either auxin signal transduction pathway or ethylene formation in plant cells were applied to cut flower stems of snapdragon (Antirrhinum majus L.) and their effects on the postharvest gravitropic response were studied. The chemical treatments were done by submerging either the stem base or the top part of cut flower, which involves the gravistimulus-sensitive region, for 1 h at $25^{\circ}C$. When the chemicals were supplied from the cut stem base, the gravitropic upward bending of flower stalks kept horizontally after the treatments with 20 mM CDTA or 10 mM $CoCl_2$ was comparable to that of the untreated control, but o-vanadate showed a certain degree of effectiveness for suppressing the bending response. In contrast, the direct application of those agents to the gravitropically sensitive region of cut flowers in the presence of 0.01% Triton X-100 resulted in a substantial reduction of the gravitropic response. In the case of 20 mM $CoCl_2$ treatment, almost total elimination of gravitropism without any significant deterioration of flower quality was observed. The results indicate the possibility of preparation of a protocol involving $CoCl_2$ and a proper surfactant for commercial use to suppress the gravitropic response of cut flowers during postharvest storage and transportation.

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Characterization of Transgenic Tall Fescue Plants Overexpressing NDP Kinase Gene in Response to Cold Stress (NDP Kinase 유전자를 과발현시킨 형질전환 톨 페스큐 식물체의 저온 스트레스에 대한 내성 특성)

  • Lee, Sang-Hoon;Lee, Ki-Won;Kim, Kyung-Hee;Yun, Dae-Jin;Kwak, Sang-Soo;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.29 no.4
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    • pp.299-306
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    • 2009
  • Oxidative stress is the main limiting factor in crop productivity. To solve global environmental problems using the plant biotechnology, we have developed on the oxidative stress-tolerant transgenic tall fescue plants via Agrobacterium-mediated genetic transformation method. In order to develop transgenic tall fescue (Festuca arundinacea Schreb.) plants with enhanced tolerance to multiple environmental stresses, nucleotide diphosphate kinase gene under the control of CaMV35S promoter were introduced into genome of tall fescue plants. Proteomic analysis revealed that transgenic tall fescue not only accumulated NDP kinase 2 protein in their cells, but also induced several other antioxindative enzyme-related proteins. When leaf discs of transgenic plants were subjected to cold stress, they showed approximately 30% less damage than wild-type plants. In addition, transgenic tall fescue plants showed normal growth when transgenic plants were subjected to $4^{\circ}C$ for 3 days treatments. These results suggest that transgene is important in ROS scavenging by induction of antioxidative proteins, and could improve abiotic stress tolerance in transgenic tall fescue plants.

The Anti-tumor Effect of Polysaccharide from the Leaves of Panax Ginseng C.A. Meyer(MB40) in a Murine Squamous Cell Carcinoma Model (편평세포암 동물 모델에서 고려인삼잎-다당체(MB40)의 항암치료효과)

  • Joo, Eun-Jung;Choi, Jun;Chung, Eun-Jae;Hong, Seok-Jin;Cho, Jae-Gu;Baek, Seung-Kuk;Woo, Jeong-Soo;Jung, Kwang-Yoon;Kwon, Soon-Young
    • Korean Journal of Head & Neck Oncology
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    • v.25 no.1
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    • pp.3-7
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    • 2009
  • Background and Objectives : Panax ginseng C.A. Meyer is a medical plant that has been widely utilized as a tonic and nutritional agent since ancient times in Korea. Ginseng has anti-metastatic property of cancer and immunomodulating activity. The novel acidic polysaccharide compound(MB40) was isolated from the leaves of Panax ginseng C.A. Meyer. To determine immunomodulating activities of MB40, we evaluate anti-cancer and anti-metastatic effects of MB40 in tumor bearing immune competent mice. Material and Methods : C3H mice were divided into three equal groups(Cisplatin treatment group, MB40 treat-ment group, Cisplatin and MB40 treatment group) and were transplanted SCC(Squamous Cell Carcinoma) cells(2${\times}$106) to the lateral side of abdomen. From day 4 after transplantation, MB40 was administrated at dose of 10mg/kg, respectively, every other day by intratumoral injection. Cisplatin was systemically administrated at doses of 1mg/kg, respectively, every week by intraperitoneal injection. Results : 5 days after administration, tumors can be palpated in every mice group. After 13 days of administration, the mice group to which MB40 were administrated exhibited reduction in tumor size respectively, compared to cisplatin group. Overall status of mice such as body weight and activity were superior in MB40 group than cisplatin group. Conclusion : The result of this study indicates MB40 may have significant therapeutic effect and decreases complications induced by systemic chemotheraphy. MB40 may be developed as a novel and potent immunotropics to improve the cell immune system and anti-cancer drug for the treatment of cancer patients in head and neck squamous cell carcinoma.

Application of Various Plants as an Inhibitor of Algal Growth: Studies in Barge Enclosure and Artificially Eutrophicated Pond (식물체를 이용한 조류제어 현장 적용성 실험)

  • Kim, Sang-Hun;Jun, Sun-Ok;Lim, Byung-Jin
    • Korean Journal of Ecology and Environment
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    • v.35 no.2 s.98
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    • pp.123-132
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    • 2002
  • Experiments have been carried out in barge enclosure near Bukhangang area and arificially eutrophicated pond. Applied plants are three terrestrial plants (Big cone pine, Pine needles and Ginkgo) and one aquatic plant (Water chestnut). In the experiment pond, applied plants are big cone pine, pine needles and ginkgo at the concentration of 0.5 g/l and water chestnut of 0.3 g/l. And it used pine needles at the concentration of 0.3 g/l in barge enclosure. The dominant species was Microcystis aeruginosa. Chlorophyll a concentration decreased to the extent of average 65% as compared with the beginning of experiment by the input of big cone pine and pine needles in artificially eutrophicated pond. But it was decreased to the extent of average 34% by the input of water chestnut. Especially, pine needles significantly inhibited the growth of algae from the early days in barge enclosure, it was declined at a rate of average 74%. It was proved that pine needles was the most inhibitory to the growth of algae at the treatment with $0.3{\sim}0.5\;g/l$, Ginkgo and water chestnut are needed more study. We purified that the dissolved phenolic compounds from the various plants can affect algal growth.

A Study of the Antioxidant Activities and Whitening Activities of Areca semen Extracts as Cosmetic Ingredient (화장품 소재로서 빈랑자의 항산화 및 미백활성에 관한 효과)

  • Kang, Hee Cheol;Cha, Mi Yeon;Kim, Jae Young
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.3
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    • pp.269-277
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    • 2015
  • Herbal plant extracts are good resources to find functional compounds for cosmetic ingredient. In this study, the extract of Areca semen (A. semen) was studied for melanogenesis inhibition and antioxidant activity. The results showed that ethyl acetate fraction of A. semen contained phenolic contents, $301.35{\pm}0.88{\mu}g/mg$, and exhibited potent antioxidant activity with $IC_{50}$ value of $1.02{\pm}0.07{\mu}g/mg$. Further, FRAP value exhibited potent antioxidant activity with $9.07{\pm}0.36mM$. Disk diffusion assay was performed for antibacterial activity. Ethyl acetate fraction of A. semen showed antibacterial activity against Staphylococcus aureus (S. aureus) and Staphylococcus epidermidis (S. epidermidis) at $80{\mu}g/mL$, whereas it showed no significant antibacterial activity against Escherichia coli (E. coli). The results of cell viability indicated that ethyl acetate fraction did not show cytotoxicity to B16/F1 cells at $80{\mu}g/mL$ and showed significant cytotoxicity at $100{\mu}g/mL$ of concentration and showed inhibition of melanin synthesis inhibitory, $29.78{\pm}0.31%$ at $80{\mu}g/mL$. Furthermore, mRNA expressions of tyrosinase and MITF were decreased after treatment with ethyl acetate fraction in a dose-dependent manner. As a result, the ethyl acetate fraction of A. semen could be considered as potential as whitening agents.

Appropriate nitrogen application enhances saponin synthesis and growth mediated by optimizing root nutrient uptake ability

  • Wei, Wei;Ye, Chen;Huang, Hui-Chuan;Yang, Min;Mei, Xin-Yue;Du, Fei;He, Xia-Hong;Zhu, Shu-Sheng;Liu, Yi-Xiang
    • Journal of Ginseng Research
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    • v.44 no.4
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    • pp.627-636
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    • 2020
  • Background: Cultivation of medicinal crops, which synthesize hundreds of substances for curative functions, was focused on the synthesis of secondary metabolites rather than biomass accumulation. Nutrition is an important restrict factor for plant growth and secondary metabolites, but little attention has been given to the plasticity of nutrient uptake and secondary metabolites synthesis response to soil nitrogen (N) change. Methods: Two year-field experiments of Sanqi (Panax notoginseng), which can synthesize a high level of saponin in cells, were conducted to study the effects of N application on the temporal dynamics of biomass, nutrient absorption, root architecture and the relationships between these parameters and saponin synthesis. Results: Increasing N fertilizer rates could improve the dry matter yields and nutrient absorption ability through increasing the maximum daily growth (or nutrient uptake) rate. Under suitable N level (225 kg/ha N), Sanqi restricted the root length and surface and enhanced the root diameter and N uptake rate per root length (NURI) to promote nutrient absorption, but the opposite status of Sanqi root architecture and NURI was found when soil N was deficient. Furthermore, increasing N rates could promote the accumulation of saponin in roots through improving the NURI, which showed a significant positive relationship with the content of saponin in the taproots. Conclusion: Appropriate N fertilizer rates could optimize both root architecture and nutrient uptake efficiency, then promote both the accumulation of dry matter and the synthesis of saponins.

Molecular genetic analysis of phytochelatin synthase genes in Arabidopsis

  • Ha, Suk-Bong
    • Proceedings of the Botanical Society of Korea Conference
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    • 2002.04a
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    • pp.62-72
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    • 2002
  • This study has investigated the biosynthesis and function of the heavy metal binding peptides, the phytochelatins, in plants. PCs are synthesised enzymatically from glutathione by the enzyme PC synthase in the presence of heavy metal ions. Using Arabidopsis thaliana as a model organism cadmium-sensitive, phytochelatin-deficient mutants have been isolated and characterised in previous studies. The cadl mutants have wildtype levels of glutathione, are PC deficient and lack PC synthase activity. Thus, the CADl gene has been proposed to encode PC synthase. The CADl gene was isolated by a positional cloning strategy The gene was mapped and a candidate identified. Each of four cadl mutants had a single base pair change in the candidate gene and the cadmium-sensitive, cadl phenotype was complemented by the candidate gene. This demonstrated the CADl gene had been cloned. A homologous gene in the fission yeast, Schizosaccharomyces pombe was identified through database searches. A targeted-deletion mutation of this gene was constructed and the mutant, like cadl mutants of Arabidopsis, was cadmium-sensitive and PC-deficient. A comparison of the redicted amino acid sequences reveals a highly conserved N-terminal region Presumed to be the catalytic domain and a variable C-terminal region containing multiple Cys residues proposed to be involved in activation of the enzyme by metal ions. Similar genes were also identified in animal species. The Arabidopsis CADl/AtPCSl and S. pombe SpbPCS genes were expressed in E. coli and were shown to be sufficient for glutathione-dependent, heavy metal activate PC synthesis in vitro, thus demonstrating these genes encode PC synthase enzymes. Using RT-PCR, AtPCSl expression appeared to be independent of Cd exposure. However, at higher levels of Cd exposure a AtPCSl-CUS reporter gene construct appeared to be more highly expressed. Using the reporter gene construct, AtPCSl was expressed most tissues. Expression appeared to be greater in younger tissues and same higher levels of expression was observed in some regions, including carpels and the base of siliques. AtPCS2 was a functional gene encoding an active PC synthase. However, its Pattern of expression and the phenotype of a mutant (or antisense line) have not been determined. Assuming the gene is functional then it has clearly been maintained through evolution and must provide some selective advantage. This implies that, at least in some cells or tissue, it is likely to be the dominant PC synthase expressed. This remains to be determined

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Anti-cancer Effects and Molecular Mechanisms of Withaferin A (Withaferin A의 다양한 항암 효과 및 분자생화학적 기전)

  • Woo, Seon Min;Min, Kyoung-Jin;Kwon, Taeg Kyu
    • Journal of Life Science
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    • v.23 no.3
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    • pp.462-469
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    • 2013
  • Withaferin A is a steroidal lactone purified from the Indian medicinal plant Withania somnifera. It exhibits a wide variety of activities, including anti-tumor, anti-inflammation, and immunomodulation properties. In this review, we focused on the anti-cancer effects of withaferin A. Withaferin A inhibits cell proliferation, metastasis, invasion, and angiogenesis in cancer cells. Furthermore, it sensitized irradiation, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-, and doxorubicin-mediated apoptosis. The results showed that multiple mechanisms were involved in withaferin A-mediated anti-cancer effects. First, withaferin A increased intracellular reactive oxygen species (ROS) production and induced ER stress- and mitochondria-mediated apoptosis. Second, withaferin A inhibited the signaling pathways (Jak/STAT, Akt, Notch, and c-Met), which are important in cell survival, proliferation, and metastasis. Third, it induced apoptosis and inhibited cancer cell migration through the up-regulation of prostate apoptosis protein-4 (Par-4). Finally, withaferin A up-regulated pro-apoptotic protein expression levels through the inhibition of proteasome activity. Our findings suggested that withaferin A is a potential, potent therapeutic agent.