• Title/Summary/Keyword: plant cultured cells

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Enhancement of eurycomanone biosynthesis in cell culture of longjack (Eurycoma longifolia) by elicitor treatment

  • Nhan, Nguyen Huu;Loc, Nguyen Hoang
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.340-346
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    • 2018
  • In this study, the effect of elicitors such as yeast extract (YE), methyl jasmonate (MeJA) and salicylic acid (SA) on the accumulation of eurycomanone in Eurycoma longifolia cell cultures were investigated. Suspension cells of E. longifolia was cultured in Murashige and Skoog (MS) medium supplemented with 30 g/L sucrose, 1.25 mg/L naphthaleneacetic acid (NAA) and 1 mg/L kinetin at a shaking speed of 120 rpm. Elicitors were added in the culture at different concentrations and times to stimulate eurycomanone accumulation in the Eurycoma longifolia cells. Eurycomanone content was determined by HPLC with a C18 column, flow rate of 0.8 mL/min, run time of 17.5 min, and a detector wavelength of 254 nm. The stationary phase was silica gel and the mobile phase was acetonitrile: $H_2O$. Non-elicited cells were used as the control. The study showed the effect of different elicitor concentrations, YE at 200 mg/L, MeJA at $20{\mu}M$ and SA at $20{\mu}M$ stimulated high production of eurycomanone. In which, treatment of $20{\mu}M$ MeJA after 4 days of culture resulted in the highest accumulation of this compound (17.36 mg/g dry weight), approximately 10-fold higher than that of untreated cells (1.70 mg/g dry weight).

Shoot Proliferation and Plant Regeneration from Suspension-Cultured Cells of Dianthus gratianopol (패랭이꽃속 Dianthus gratianopol의 현탁배양세포로부터 Shoot 증식과 식물체 재분화)

  • Kim Joon-Chul
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.301-306
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    • 2005
  • Conditions for efficient organogenesis and plant regeneration from Dianthus gratianopol suspension cultured cells were established. Shoot-forming calli of glossy surface, pale green and knobby type were selected from leaf explant-derived calli and were suspension-subcultured every week in CP liquid medium with 1.0 mg/L 2,4-D and 0.5 mg/L BAP. Combinations of 1.0 mg/L 2,4-D and 0.5 mg/L BAP, and 1.5 mg/L 2,4-D and 0.5 mg/L BAP were effective for the induction of regenerative callus from the suspension cultured cell clusters. Multiple shoot primordia were initiated from the green spots of these regenerative callus and formed shoots on MS medium with 1.0 mg/L TDZ and 0.5 mg/L PAA. Shoot regeneration frequency (calli regenerating at least one shoot) was about 87%. For plant regeneration, proliferated shoots were excised and transferred to MS medium with 0.1 mg/L NAA for root initiation after 9 weeks of culture. The regenerants were potted in soil and formed the flowering buds and petals. Also, adventitious shoots were formed from the excised green shoot primordia of regenerative callus and these shoots proliferated successfully and regenerated to whole plants.

Molecular Cloning and Characterization of a Peroxiredoxin cDNA from Cell Cultures of Sweetpotato (고구마 배양세포에서 Peroxiredoxin cDNA의 분리 및 발현 특성)

  • Park, Soo-Young;Ryu, Sun-Hwa;Kwon, Suk-Yoon;Kim, Jong-Guk;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.135-141
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    • 2003
  • Peroxiredoxin(Pix) are large family of peroxidases that reduce alkyl hydroperoxides and hydrogen peroxide. A cDNA clone (referred to as swPrxl) encoding Pix was from a sweetpotato cDNA library constructed from suspension-sultured cells, and its expression was investigated in terms of stress. The swPrxl contained an open reading frame (ORF) encoding mature protein of 193 amino acids with calculated molecular mass of 20.8kDa. The predicted amino acid sequence of swPrxl has two conserved cysteines that are essential resicues for the reduction of peroxides. It showed high amino acid sequence homology ot PixIIF of Arabidopsis (77%) and putative Prx of rice(72%). RNA gel-blot analysis showed that swPrxl gene was expressed dominantly in leave among intact tissues, and also highly detect in suspension-cultured cells. Interestingly, the level of swPrxl transcripts was almost the same regardless of the growth stage in suspension culture. Furthermore, the transcription level of swPrxl gene was not significantly changed in response to various stress treatments such as wounding, extreme temperature and stress-related chemicals RT-PCR analyses.

Inhibition of mTOR signaling pathway by aqueous extract of Siberian ginseng

  • Byun, Boo Hyeong;Cho, Tae Hwan;Park, Kyeong Mee
    • The Journal of Korean Medicine
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    • v.38 no.2
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    • pp.7-14
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    • 2017
  • Objectives: This study evaluated the effect of aqueous extract from roots of Siberian ginseng on mTORC1 pathway. Methods: mTORC1 activity was measured by the phosphorylation status of p70 S6 kinase (S6K) in HeLa cells as well as the brain, liver and muscle tissues in diabetic db/db mice. Autophagy induction after the treatment of Siberian ginseng extract was evaluated by monitoring the conversion of cytoplasmic LC3I into lipidated LC3II in cultured human HeLa GFP-LC3 cells. Cell cycle analysis was performed in HeLa cells treated with Siberian ginseng using flow cytometry. Results: Among >2,800 plant products used for oriental medicine, Siberian ginseng was found to inhibit mTORC1 to phosphorylate S6 kinsase (S6K) in HeLa cells as well as the brain, liver and muscle tissues in diabetic db/db mice. Siberian ginseng-mediated mTORC1 activity was reversible unlike the prolonged suppression of mTORC1 by rapamycin when HeLa cells were grown in fresh media after the removal of the inhibitors. Siberian ginseng extract at concentrations to inhibit mTORC1 was not overly cytotoxic in cultured HeLa cells whereas rapamycin was obviously cytotoxic. The conversion of cytoplasmic LCI into lipidated LCII was increased by fivefold in HeLa GFP-LC3 cells treated with Siberian ginseng extract. Progression of cell cycle was attenuated at G2/M phase by the treatment of Siberian ginseng extract. Conclusions: These results suggest that the aqueous extract of Siberian ginseng possibly plays a good therapeutic role in various diseases involving mTORC1 signaling.

Production of Polyhydroxybutyrate from Crude Glycerol and Spent Coffee Grounds Extract by Bacillus cereus Isolated from Sewage Treatment Plant

  • Lee, Gi Na;Choi, So Young;Na, Jonguk;Youn, HaJin;Jang, Yu-Sin
    • KSBB Journal
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    • v.29 no.6
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    • pp.399-404
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    • 2014
  • Production of biodegradable polymer polyhydroxyalkanoates (PHAs) from industrial wastes exhibits several advantages such as recycle of waste and the production of high valuable products. To this end, this study aimed at isolating from the sewage treatment plant a PHA producing bacterium capable of utilizing wastes generated from biodiesel and food industries. A Bacillus cereus strain capable of producing poly(3-hydroxybutyrate) [P(3HB)] was isolated, which was followed by confirmation of P(3HB) accumulation by gas-chromatographic analyses. Then, the effects of nutrient limitation on P(3HB) production by B. cereus was first examined. Cells cultured in a minimal medium under the limitation of nitrogen, potassium and sulfur suggested that nitrogen limitation allows the highest P(3HB) accumulation. Next, production of P(3HB) was examined from both waste of biodiesel production (crude glycerol) and waste from food industry (spent coffee grounds). Cells cultured in nitrogen-limited minimal medium supplemented crude glycerol and waste spent coffee grounds extract accumulated P(3HB) to the contents of 2.4% and 1.0% of DCW. This is the first report demonstrating the capability of B. cereus to produce P(3HB) from waste raw materials such as crude glycerol and spent coffee grounds.

Cell Viability and Antioxidant Enzyme Activity in the Cell of Ginseng (Panax ginseng C.A. Meyer) Treated with Soil Extracts (인삼재배지의 토양추출물이 종자 발아와 세포의 항산화효소 활성에 미치는 영향)

  • Ryu, Tae-Seok;Kwon, Soon-Tae
    • Korean Journal of Plant Resources
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    • v.21 no.4
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    • pp.324-328
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    • 2008
  • One hundred-eighty extracts of soil collected from ginseng (Panax ginseng C.A. Meyer) fields were subjected to lettuce germination test, electrolyte leakage, cell viability and antioxidant enzyme activity test. Regardless of various cultivation periods, there was no significant difference in soil pH, the content of organic matter and available phosphate in ginseng fields. Based on lettuce seed germination test, six soil extracts showing inhibition of germination and/or seedling growth were selected for further study. Selected soil extracts markedly inhibited cell viability of ginseng cultured cells but leakage of electrolytes were not affected by the treatment. Enzyme activity of superoxide dimutase in ginseng cultured cells was not affected by the treatment with the soil extracts. However, those of peroxidase and catalase were significantly inhibited by the treatment with soil extracts which showed inhibition of lettuce seed germination and seedling growth.

Inhibitory Effect of Microcystis aeruginosa (Cyanophyceae) Growth by Plants in vitro (식물체를 이용한 조류증식억제 효과)

  • Jheong, Weon-Hwa;Byeon, Myeong-Seop;Jun, Sun-Ok;Lim, Byung-Jin
    • Korean Journal of Ecology and Environment
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    • v.33 no.2 s.90
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    • pp.136-144
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    • 2000
  • M. aeruginosa isolated from Lake Paldang was cultured in CB medium, and then each wet plants put into the cultured medium at a rate of 0.5 g and 2.5 g wet wt/l. There was slight inhibition by the input of cattail and iris of each 0.5 g wet wt/l cultured medium, but showed no reduction in algal growth in other flasks. Among the applied plants, ginkgo, pine needles, big cone pine, waterreed and water chestnut had an effect on inhibition of algal growth at the input of 2.5 g wet wt/l. Plants which were dried for 3 days at $50^{\circ}C$ introduced into the testing flask for 10days cultured at dose rates of 2.5 g/l. When chlorophyll a concentration was remarkably high as $802.6\;{\mu}g/l$ after five days, there was noticeably less chlorophyll compared with control at a rate of 98% by big cone pine, 96% by ginkgo, 95% by pine needles and 86% by rice straw, respectively. To examine the effect of plant extracts on algal growth, big cone pine and water chestnut were put to the amount of 1.25 g liquid extracts/l. Chlorophyll a concentration and cell density decreased to the extent of average 43% as compared with the beginning of experiment, but when concentration of chlorophyll a increased a most high, the inhibition of algal growth by liquid extracts did not affect at all. When a quantity of plant equivalent to 2.5 g liquid extracts/l inhibited the growth of algae by 95% after nine days.

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Effects of Several Medicinal Plants Extract on Survival Rate, Chlorophyll Contents and Photosynthetic Electron Transport Activity of Liverwort Photoautotrophic Cultured Cell (약용식물 추출액이 우산이끼 자가관양배양세포의 생존율, 엽록소함량 및 광합성전자전달 활성에 미치는 영향)

  • 정형진;권순태;김시무
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.2
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    • pp.133-141
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    • 1995
  • The effects of allelochemicals from medicinal plants have been studied as photo-synthetic inhibitor for photoautotrophic(PA) cultured cells. The extracts from 9 plant species were used for measuring the physiological effects on the liverwort cultured cell in following areas; germination inhibition, chlorophyll contents, hill activity, cell viability, photosynthetic oxygen evolution,and protein pattern changes on SDS PAGE. Germination inhibitions were detected in all plant after treating with 10% extract. Especially, treatment with 10% extract from Pulsatilla koreana and Aconitum carmichael inhibited germinations completely. Chlorophyll fornation was inhibited completely by treating PA cells with extract of Pulsatilla koreana, whose effect was similar to that of DCMU 10-3M, inhibitor for photosynthetic electron trans-fer. The treatment with extract from Pulsatilla koreana on PA cell showed the highest hill activity and the lowest cell viability among extracts studied. Oxygen releasing has been decreased down to 14-77% after treating with extracts from Pinellia ternata, Araliacont inentaila, Pulsatilla koreana and Vitex rotundifolia. Especially, 60$\mu$l of Pulsatilla koreana extract into 2ml mixture of PA cell inhibit-ed oxygen release up to 50%. Protein bands on SDS-PAGE, 14kD, 31kD, 41kD, 53kD, and 73kD, were not detected after treating Pulsatilla koreana extract on PA cells.

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THE STUDY ON TISSUE CULTURED WILD MOUNTAIN GINSENG(Panax Ginseng C.A. Meyer) ADVENTITIOUS ROOTS EXTRACT AS A COSMETIC INGREDIENT

  • Jung, Eun-Joo;Park, Jong-Wan;Kim, Joong-Hoi;Paek, Kee-Yoeup
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.611-616
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    • 2003
  • Korean ginseng(Panax Ginseng C.A. Meyer) known as a oriental miracle drug is an important medicinal plant. Ginseng has been used for geriatric, tonic, stomachic, and aphrodisiac treatments for thousands years. Also, it is an antibiotic and has therapeutic properties against stress and cancer. Ginseng is widely distributed all over the world. Among them, Korean mountain ginseng has the most valuable effect on pharmaceuticals. The roots of mountain ginseng contained several kinds of ginsenosides that have many active functions for the human body. However, the study of mountain ginseng has a limit because the mountain ginseng is very expensive and rare. So, we artificially cultured mountain ginseng adventitious roots using the bioreactor culture system. We induced callus from original mountain ginseng, directly dug up in mountain and aged about one hundred ten years. Separated adventitious roots were precultured in 500ml conical flasks and then, transferred in 20L bioreactors. The adventitious roots of mountain ginseng were harvested after culturing for 40days, dried and then, extracted with several solvents. In this study, we investigated the whitening effect, anti-wrinkle effect and the safety of tissue cultured adventitious roots extract of mountain ginseng in order to identify the merit as a cosmetic ingredient. Particularly, extract of mountain ginseng adventitious roots showed whitening and anti-wrinkle effects. The inhibitory effect of this extract on the melanogenesis was examined using B-16 melanoma cell. When B-16 melanoma cells were cultured with adventitious root extract, there was a dramatically decrease in melanin contents of 8-16 melanoma cell. And we identified this extract inhibited Dopa auto-oxidation significantly. Also, when transformed mouse fibroblast L929 cells were treated with this extract, there was a significant increase in collagen synthesis. The results show significant inhibited melanization and wrinkle without inhibiting cell viability.

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Effect of Calcium Ion on Mesophyll Protoplast Culture of Arabidopsis thaliana (Arabidopsis thaliana의 엽육세포 원형질체배양에 미치는 칼슘이온의 영향)

  • 박현용
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.5
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    • pp.277-281
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    • 1995
  • The present study was performed to investigate the effect of calcium ion on the mesophyll protoplast culture of Arabidopsis thaliana. The mesophyll protoplase were isolated and cultured on an IMH medium supplemented with CaCl$_2$of various concentrations. When the protoplasts were cultured on the medium containing 0 to 12.5 mM CaCl$_2$, extreme vacuolization occurred without cell division. When the protoplasts were cultured with higher levels of CaCl$_2$ up to 50 mM, vacuolization decreased dose-dependent): and the number of plasma-rich cells increased. Cell division was induced when the protoplast were cultured on the medium with CaCl$_2$ higher than 25 mM. The highest plating efficiency (5-6%) was obtained with 50 mM CaCl$_2$. However the plating efficiency was markedly inhibited by 100mM CaCl$_2$ or above. These resole suggest that the relatively high concentration (50 mM) of calcium ion may be required for the culture of protoplasts.

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