• Title/Summary/Keyword: Root induction

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Molecular Characterization of Cinnamate 4-Hydroxylase gene in Red Hot Pepper (Capsicum annuum L.) (고추에서 분리한 Cinnamate 4-Hydroxylase 유전자의 분자생물학적 특성)

  • Kim Kye-Won;Ha Sun-Hwa;Cho Kang-Jin;Kim Eun-Ju;Lee Min-Kyung;Yu Jae-Ju;Kim Jong-Guk;Lee Shin-Woo
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.167-173
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    • 2005
  • Three different cDNAS for cinnamate 4-hydroxylase (C4H) which are involved in the second step of the general phenylpropanoid pathway were isolated and designated as pc4h1 (1,755 bp), pc4h2 (1,655 bp), and pc4h3 (1,316 bp), respectively. The nucleotide sequence analysis revealed that both pc4h1 and pc4h2 clones encode polypeptides of 505 amino acids frame but pc4h3 clone was truncated at the 5'-end of coding region. The alignment of the deduced amino acid sequences showed that PC4H1 and PC4H2 are highly homologous (95.8% identical) with each other and contain three conserved domains which are typical in cytochrome P450 monooxygenase: proline-rich region, threonine-containing binding pocket for the oxygen molecule, and heme binding region. In addition, result of the phylogenic tree analysis revealed that both pepper C4Hs belong to Class 1. pc4h2 transcription was strongly induced in wounded fruit (400%) and root (200%) relative to its very low basal level but not in leaf or stem tissue. In case of pc4h1, the basal level of transcription was higher than pc4h2 but induction by wounding was lower in fruit and root while leaf and stem tissues did not respond to wounding. The basal level of pc4h3 transcripts was not, if any, detectable and response to wounding was not observed.

Antioxidative and Biological Activites of Extracts of Sweetpotato Tips (고구마 끝순 추출물의 항산화 및 생리활성)

  • Lee, Joon-Seol;Park, Yang-Kyun;Ahn, Young-Sup;Kim, Hag-Sin;Chung, Mi-Nam;Jeong, Byeong-Choon;Bang, Jin-Ki
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.2
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    • pp.228-238
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    • 2007
  • This study was conducted to increase sweetpotato utilization and to determine the vegetative value of sweetpotato tips by investigating the phenolic compounds, antioxidative effect in oil, electron donating ability, nitrite scavenging effect and ACE inhibition activities. The phenolic compounds present in sweetpotato tips are the gallic, chlorogenic, gentisic, caffeic, couramic and ferulic acid, which are 16-122 times higher compared to other vegetables such as spinach, soybean sprout, and perilla leaves. In each solvent extract, the total phenolic compounds (175.8mg/g) was composed of 55% EtOAc extraction and 39% BuOH extract, respectively. The results of induction period using the Rancimat method showed that the antioxidant activity of SP tips was higher than the tocopherol or BHT. The relative levels of each solvent extract in SP tips were as follows: EtOAc>BHT>BuOH>Tocopherol>Water>$CHCl_3$>Hexane. The peroxide value was measured every 5 days for 25 days during storage and results showed that the peroxide value, the tips, tuberous root and tocopherol were lower compared to spinach, soybean sprout and perilla leaves. Nitrite scavenging effects were excellent in sweetpotato tips, perilla leaves and soybean sprout, especially, inhibition rate of perilla leaves (72%) were superior to the others. In process of solvent extraction, activity of BuOH and water extractions were the best. ACE inhibition activity in sweetpotato tips was 1.5 times higher than in tuberous roots and $1.9{\sim}3.7$ times higher than in spinach, soybean sprout, perilla leaves.

Effects of Adenophora triphylla Ethylacetate Extract on mRNA Levels of Antioxidant Enzymes in Human HepG2 Cells (인간 HepG2 Cell에서 항산화 효소의 mRNA 발현에 대한 잔대 에틸아세테이트 추출물 효과)

  • Choi, Hyun-Jin;Kim, Soo-Hyun;Oh, Hyun-Taek;Chung, Mi-Ja;Cui, Cheng-Bi;Ham, Seung-Shi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.10
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    • pp.1238-1243
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    • 2008
  • The root of Adenophora triphylla is widely used as traditional herbal medicine in Korea. We studied its effects on sodium nitroprusside (SNP) cytotoxicity and antioxidant genes expression in HepG2 cells. To study whether Adenophora triphylla ethylacetate extract (ATea) inhibited NO-induced cell death, HepG2 cells were preincubated for 24 hr with 50 and 100 $\mu$g/mL ATea followed by 24-hr exposure to 0.5 mM SNP (exogenous NO donor). No-induced cytotoxicity was inhibited by pretreatment of ATea, as assessed by mitochondrial dehydrogenase activity (MTT assay). We further investigated the effects of ATea on mRNA levels of various enzymes of the antioxidant system such as Cu, Zn superoxide dismutase (SOD 1), Mn SOD (SOD 2), glutathione peroxidase (GPx), catalase and several enzymes of the glutathione metabolism [glutathione reductase (GR), $\gamma$-glutamyl-cystein synthetase (GCS), glutathione-S-transferase (GST), $\gamma$-glutamyltranspeptidase ($\gamma$-GT), glucose-6-phosphate dehydrogenase (G6PD)] by RT-PCR. CAT, GCS, GR and G6PD mRNA levels were increased after treatment with ATea. The SOD 1, SOD 2, GPx, GST and $\gamma$-GT mRNA levels were not affected in ATea-treated HepG2 cells. We concluded that ATea have an indirect antioxidant effects, perhaps via induction of CAT, GCS, GR and G6PD.

Antioxidative and Biological Activites of Extracts of Sweetpotato Tips (고구마 끝순 추출물이 알코올 투여 흰쥐의 항산화 효소계 및 지질과산화에 미치는 영향)

  • Lee, Joon-Seol;Park, Yang-Kyun;Ahn, Young-Sup;Kim, Hag-Sin;Chung, Mi-Nam;Jeong, Byeong-Choon;Bang, Jin-Ki
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.4
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    • pp.411-420
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    • 2007
  • This study was conducted to increase sweetpotato utilization and to determine the vegetative value of sweetpotato tips by investigating the phenolic compounds, antioxidative effect in oil, electron donating ability, nitrite scavenging effect and ACE inhibition activities. The phenolic compounds present in sweetpotato tips are the gallic, chlorogenic, gentisic, caffeic, couramic and ferulic acid, which are 16-122 times higher compared to other vegetables such as spinach, soybean sprout, and perilla leaves. In each solvent extract, the total phenolic compounds(175.8 mg/g) was composed of 55% EtOAc extraction and 39% BuOH extract, respectively. The results of induction period using the Rancimat method showed that the antioxidant activity of SP tips was higher than the tocopherol or BHT. The relative levels of each solvent extract in SP tips were as follows: EtOAc>BHT>BuOH>Tocopherol>Water>$CHCl_3$>Hexane. The peroxide value was measured every 5 days for 25 days during storage and results showed that the peroxide value, the tips, tuberous root and tocopherol were lower compared to spinach, soybean sprout and perilla leaves. Nitrite scavenging effects were excellent in sweetpotato tips, perilla leaves and soybean sprout, especially, inhibition rate of perilla leaves(72%) were superior to the others. In process of solvent extraction, activity of BuOH and water extractions were the best. ACE inhibition activity in sweetpotato tips was 1.5 times higher than in tuberous roots and $1.9{\sim}3.7$ times higher than in spinach, soybean sprout, perilla leaves.

Pro-apoptotic Effects of Platycodin D Isolated from Platycodon grandiflorum in Human Leukemia Cells (도라지 유래 사포닌 platycodin D에 의한 인체 백혈병세포의 apoptosis 유도)

  • Park, Sang Eun;Lee, Su Young;Shin, Dong Yeok;Jeong, Jin-Woo;Jin, Myung Ho;Park, Seon Young;Chung, Yoon Ho;Hwang, Hye Jin;Hong, Sang Hoon;Choi, Yung Hyun
    • Journal of Life Science
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    • v.23 no.3
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    • pp.389-398
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    • 2013
  • Platycodin D is a major constituent of triterpene saponins, which is found in the root of Platycodon grandiflorum, Platycodi Radix, which is widely used in traditional Oriental medicine for the treatment of many chronic inflammatory diseases. Several pharmacological effects of this compound have been reported recently, such as anti-inflammation, immunogenicity, anti-adipogenesis, lowered cholesterol, and anti-cancer activity. However, the mechanism by which this action occurs is poorly understood. In this study, we found that platycodin D greatly increased the potential of the anti-proliferative effect in various cancer cell lines. Our data revealed that platycodin D treatment resulted in a time- and concentration-response growth inhibition of U937 cells by inducing apoptosis, as evidenced by the formation of apoptotic bodies, chromatin condensation, and the accumulation of cells in the sub-G1 phase. Apoptosis induction of U937 cells by platycodin D correlated with an increase in the Bax/Bcl-2 ratio and caused the down-regulation of IAP family members. In addition, platycodin D treatment resulted in proteolytic activation of caspase-3, the concomitant degradation of poly(ADP-ribose) polymerases, and the collapse of the mitochondria membrane potential (${\Delta}{\Psi}_m$). However, the cytotoxic effects induced by platycodin D treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, which demonstrated the important role that caspase-3 played in the observed cytotoxic effect. These findings suggest that platycodin D may be a potential chemotherapeutic agent for use in the control of human leukemia U937 cells. These findings also provided important new insights into possible molecular mechanisms of the anti-cancer activity of platycodin D.

The Establishment of Nitrogen Fixation by Cultured Cell-Rhizobium Association Through Tissue Culture Technique in Soybean (대두조직배양세포(大豆組織培養細胞) - Rhizobium에 의(依)한 질소고정력(窒素固定力))

  • Kang, Sang Jae;Park, Woo Churl
    • Current Research on Agriculture and Life Sciences
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    • v.4
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    • pp.27-35
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    • 1986
  • This experiment was carried out to elucidate the factor of nitrogenase formation and to establish the nitrogen fixation system in mixed culture of cultured cells and rhizobia through tissue culture technique using three soybean varieties, Hwangkeum, Namcheon and D 68-0099 as host plants. The results obtained were as follows; The callus was induced in embryo and radicle, but not in hypocotyl. The most favorable callus induction was caused by the individual application of 2,4-D and NAA at the concentration of 2mg/1 and 4mg/1, respectively, but in case of treating both 2,4-D and kinetin, that was done at the concentration of 0.2mg(2,4-D)/0.05mg(kinetin)per liter. The growth of cultured cell was good at the concentration of 2.0mg(2,4-D)/1 and 0.2mg(2,4-D)/0.05mg(kinetin)per liter. When cultured cells were inoculated with R. japonicum 019 and 011, their growthes were considerably inhibited. The addition of single amino acid inhibited the growth of cultured cells. Hwangkeum was inhibited considerably by methionine and leucine. The inhibition of growth by single amino acid can be abolished by the addition of certain amino acids. The differentiation of adventitious root was good at the concentration of 2.0mg 2,4-D and 0.2mg 2,4-D/0.05mg kinetin per liter. Of three host plants tested with 25 R. japonicum strains, Hwangkeum had affinity for 10 strains, Namcheon for 7 strains and D68-0099 for none. The nitrogen fixing abilities of Hwangkeum and Namcheon caused by cultured cell-Rhizobium association were high in strain 019, 007, and in 007 mixed with 119, respectively.

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Anti-proliferation, Cell Cycle Arrest, and Apoptosis Induced by Natural Liquiritigenin from Licorice Root in Oral Squamous Cell Carcinoma Cells (구강편평세포암종 세포에서 감초 유래 Liquiritigenin의 항증식, 세포주기 정지 및 세포사멸 유도)

  • Kwak, Ah-Won;Yoon, Goo;Chae, Jung-Il;Shim, Jung-Hyun
    • Journal of Life Science
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    • v.29 no.3
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    • pp.295-302
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    • 2019
  • Liquiritigenin (LG) is a chiral flavonoid isolated from the roots of licorice. It exhibits multiple biological activities including anti-oxidant, anti-cancer, and anti-inflammatory effects. In particular though, the anti-cancer activity of LG in oral squamous cell carcinoma has yet to be elucidated, and LG-induced apoptosis in oral squamous cell carcinoma remains poorly understood. In the present study, we tested the role of LG in inducing apoptosis in oral squamous cell carcinoma cells. LG treatment of HN22 cells resulted in a dose-dependent inhibition of cell viability as detected by a 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay. The induction of apoptosis in terms of Annexin V/7-Aminoactinomycin D staining, sub-G1 population, and multi-caspase activity were assessed with a $Muse^{TM}$ Cell Analyzer. Flow cytometric analysis revealed that LG treatment resulted in G2/M arrest in cell cycle progression and downregulation of cyclin B1 and CDC2 expression in a concentration-dependent manner. It also resulted in significant upregulation of p27. In addition, LG was seen to trigger the generation of reactive oxygen species and induce CCAAT/enhancer-binding protein homologous protein and 78-kDa glucose-regulated protein in concentration-dependent upregulation. The LG treatment of HN22 cells led to a loss of mitochondrial membrane potential (${\Delta}{\Psi}m$); it also reduced the levels of anti-apoptotic protein and increased the expression of apoptotic protease activating factor-1, cleaved poly (ADP-ribose)polymerase and Bax. Overall, our results indicate that the pro-apoptotic effects of LG in HN22 cells depend on the activation of both intrinsic and extrinsic signaling pathways. Thus, our results suggest that LG constitutes a natural compound with a potential role as an anti-tumor agent in oral squamous cell carcinoma.

In vitro Multiplication of Hosta Tratt. Species Native to Korea by Shoot-tip Culture (경정배양에 의한 한국 자생 비비추속 식물의 기내증식)

  • Choi, Han;Yang, Jong Cheol;Ryu, Sun Hee;Yoon, Sae Mi;Kim, Sang Yong;Lee, Seung Youn
    • Korean Journal of Plant Resources
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    • v.32 no.1
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    • pp.53-62
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    • 2019
  • The purpose of this study was to establish the in vitro propagation system by shoot tip culture of six Hosta species native to Korea (Hosta capitata (Koidz.) Nakai, H. clausa Nakai, H. jonesii M.G.Chung, H. minor (Baker) Nakai, H. venusta F.Maek., and H. yingeri S.B.Jones) for mass proliferation and a new cultivar development. The shoot tips of each Hosta species were cultured on MS medium containing eight combinations of 0.5, 1.0, 2.0, 4.0 mg/L BA with 0.1 mg/L NAA, 0.1, 0.5, 1.0, 2.0 mg/L TDZ with 0.1 mg/L NAA, and without any PGRs (control). They were investigated on callus, somatic embryo, crown bud, differentiation and growth of shoot and root, total fresh weight after 8 weeks of culture. In all six Hosta species, callus and somatic embryo induction rate and multiple shooting rate of the PGRs treatment group were higher than that of the control group. The highest number of differentiated shoots were obtained on medium supplemented with 2.0 ㎎/L TDZ in H. capitata (5.4), 1.0 mg/L TDZ in H. clausa and H. jonesii (3.3 and 5.8, respectively), 0.5 mg/L BA in H. minor (11.1), 1.0 mg/L BA and 0.1 mg/L TDZ in H. venusta (8.1), and 0.5 mg/L TDZ in H. yingeri (9.8). In somatic embryo formation, the PGRs treatment group of H. jonesii and H. yingeri were more effective than the control group, and the effects were relatively less in H. capitata, H. clausa Nakai, H. minor, H. venusta. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. H. clausa showed no significant effect on callus and shoot differentiation regardless of the type and concentration of cytokinin, but slightly increased in formation of crown bud in TDZ.

Transition of Rice Culture Practices during Chosun Dynasty through Old References V. Cultivation and Cropping Patterns (주요 고농서를 통한 조선시대의 도작기술 전개 과정 연구 V. 재배양식)

  • Lee, Sung-Kyum;Guh, Ja-Ok;Lee, Eun-Woong;Lee, Hong-Suk
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.37 no.1
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    • pp.104-115
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    • 1992
  • The rice culture techniques included 'Jodosukyungbeob'(旱稻水耕法 : culture techniques of early-ripening paddy rice), 'Mandosukyungbeob' (晩稻水耕法) : culture techniques of late-Ripening paddy rice 'Handobeob'(旱稻<山稻>法 : culture techniques of upland rice), 'Myojongbeob'(苗種法 : culture techniques of paddy rice by transplanting), 'Kunangbeob'(乾秧法 : culture techniques of rice by transplanting which rears seeding in dry paddy) and 'Sudogunpanongbeob'(水稻乾播農法 : culture techniques of paddy rice seeding in dry field). Especially, 'Kunangbeob' and 'Sudogunpanongbeob' were originally developed in Korea as seen in 1600s(Kyoungje : 經濟) and early 1800s (Yoji : 要旨). In 'Jodosukyungbeob' it took 9 days for seed dipping, water-sprouting and prevent damage by birds, each for 3 days in China, but in Korea seed dipping in water took 3 days and the rest of the procedures were flexibly established. In matured soils, practices were fall plowing right after harvest, recognition of effective tillering and additional fertilization use of human manure, and stimulation of sprouting by lime application. The unique culture techniques adequate for Korean situations were practiced, which included weed control after draining accurately for 3 to 4 times, draining at mid season for improving wind and drought tolerance, rice harvesting at appropriate time for preventing grain shattering, and seeding in rows. 'Mandosukyungbeob' was improved techniques contrast to those of China, and the major contents were selection of proper varieties, good stand establishment by seeding high rates, induction of vigorous tillers, and adoption of 'Jokjongbeob'(足種法 : seeding method by foot). Also, one of the most prominent rice cultures by our ancestors was 'Kunpanongbeob' that was systemized form habitual practice of Pyongan Province. The unique technique actualized was 'Hando [旱稻(山稻)]' culture technique which was the combinations of 'Jokjongbeob', root stimulation method, and disaster-tolerant mixture cropping with adoptation of variety theory, although it was originated from China. The transplanting techniques has come before 'Jikseol'($\ulcorner$直說$\lrcorner$) and its merits were sufficiently realized. However, this method was basically prohibited from the early Chosun dynasty because extremely bad harvest was expected under drought conditions and insufficient conditions of water storage. But, it was permitted in the areas that contained water all the times and in case of large-scale farming especially. Most of rice culture was transplanted in the end of the Chosun dynasty because transplanting was continuously spreaded in the three southern provinces of Korea. Under these circumstances, transplanting technique was improved from the early to the end of the Chosun dynasty by weed control, fertilizing, water management, and quadratic transplanting. Based on these techniques, agricultural productivity was improved 5 times by that time. 'Kunpanongbeob' was created and developed properly for Korean conditions that is dry in early season and flooding in late season. This was successively developed and established into transplanting technique of nursery seedling.

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Curcumin-induced Cell Death of Human Lung Cancer Cells (Curcumin에 의해 유도되는 인간 폐암 세포주의 세포사멸)

  • Hwasin Lee;Bobae Park;Sun-Nyoung Yu;Ho-Yeon Jeon;Bu Kyung Kim;Ae-Li Kim;Dong Hyun Sohn;Ye-Rin Kim;Sang-Yull Lee;Dong-Seob Kim;Soon-Cheol Ahn
    • Journal of Life Science
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    • v.33 no.9
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    • pp.713-723
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    • 2023
  • Lung cancer is a type of cancer that has the highest mortality rate. It is mainly classified into small-cell lung cancer (SCLC) and non-small-cell lung cancer (NSCLC). Chemotherapy is used to treat lung cancer, but long-term treatment causes side effects and drug resistances. Curcumin is a bright yellow polyphenol extracted from the root of turmeric. It has biological activities, such as anti-oxidant, anti-cancer, and anti-inflammatory effects. In this study, we observed differential cell death in human lung cancer cells. Based on the results, curcumin at 10, 30, and 50 μM exhibited a dose-dependent inhibition on the cell survival of several lung cancer cells, with minor differential phenotypes. In addition, apoptosis, autophagy, and reactive oxygen species (ROS) regeneration were observed through flow cytometry. Curcumin dose-dependently increased these phenotypes in A549 (NSCLC) and DMS53 (SCLC), which were restored by corresponding inhibitors. Western blotting was performed to measure the level of expression of apoptosis- and autophagy-related proteins. The results indicate that Bax, PARP, pro-caspase-3, and Bcl-2 were dose-dependently regulated by curcumin, with seemingly higher Bax/Bcl-2 ratios in DMS53. In addition, autophagic proteins, p-AKT, p62, and LC3B, were dose-dependently regulated by curcumin. ROS inhibition by diphenyleneiodonium reduced the induction of apoptosis and autophagy generated by curcumin. Taken together, it is suggested that curcumin induces apoptosis and autophagy via ROS generation, leading to cell death, with minor differences between human lung cancer cells.