• 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).

• Korean Journal of Medicinal Crop Science
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• v.13 no.3
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• pp.69-76
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• 2005

The young leaves of A. princeps have been a well known for a crude medicine and used in treatment of colic pain, vomiting and menstrual irregularity. Based on TLC and HPLC and used an artemisinin, an anti-malarial compounds which is believed to be detected only in A. annuaup so far can be biosynthesized in A. princeps. To investigate the production of secondary metabolites like artemisinin in cultured cells, the cell culture of A. princeps was established. Callus and suspension cultured cells of A. princeps were induced and grown highest in MS media containing $0.2\;mg/{\ell}$ 2,4-D, $0.1\;mg/{\ell}$ BAP and 2% sucrose. Different metabolites from in vitro cultured cells (callus and suspension cultured cell) and intact plants were analyzed by TLC analysis. As a result, we can confirm that in vitro culture has a potential for mass production of secondary metabolites from A. princeps.

• Plant Biotechnology Reports
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• v.4 no.2
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• pp.125-128
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• 2010

Culture conditions were established for high frequency plant regeneration via somatic embryogenesis from cell suspension cultures of Nymphoides coreana. Zygotic embryos formed pale-yellow globular structures and calluses at a frequency of 85.6% when cultured on half-strength Murashige and Skoog (MS) medium supplemented with 0.3 $mg\;l^{-1}$ of 2,4-D. However, the frequency of pale-yellow globular structures and white callus formation decreased slightly with an increasing concentration of 2,4-D up to 10 $mg\;l^{-1}$ with the frequency rate falling to 16.7%. Cell suspension cultures were established from zygotic embryo-derived calluses using half-strength MS medium supplemented with 0.3 $mg\;l^{-1}$ of 2,4-D. Upon plating onto half-strength MS basal medium, over 92.3% of cell aggregates gave rise to numerous somatic embryos and developed into plantlets. Regenerated plantlets were successfully transplanted into potting soil and achieved full growth to an adult plant in a growth chamber. The high frequency plant regeneration system for Nymphoides coreana established in this study will be useful for genetic manipulation and cryopreservation of this species.

• Proceedings of the Zoological Society Korea Conference
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• 1999.10b
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• pp.208.1-208
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• 1999

No Abstract, See Full Text

• Korean Journal of Plant Tissue Culture
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• v.25 no.6
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• pp.477-485
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• 1998

To underatand in vitro regulation of differentiation, the effects of growth regulators and nitrogen source on metabolism of cell wall polysaccharides in suspension culture of kidney bean (Phaseolus vulgaris L.) were investigated. The suspension cells (cell clusters) were directly induced from the epicotyl segments of the seedlings, which were cultivated in MS medium supplemented with 1.0mg/L of 2,4-D and 0.5 mg/L of kinetin. When compared with cell wall sugar contents of the epicotyl segments, the cellulose content of the suspension-cultured cells decreased; while the pectin and hemicellulose content increased; suggesting increases of rhamnogalacturonan I and arabinogalactan IIduring the dedifferentiation, respectively, The effects of growth regulators(2,4-D, 1.0mg/L and kinetin, 0.5mg/L) and nitrogen source (potasium nitrate, 19.0mg/L and ammonium nitrate, 16.5 g/L) in the medium on the proliferation and the turnover of the cell wall polysaccharides were investigated for 30 days. In the medium with growth regulators and without nitrogen source, the proliferation rate was extremely high (16 folds). Growth regulators and nitrogen source increased the pectin content. Analysis of neutral sugar composition of pectin fraction showed that nitrogen source enhanced rhamnose level remarkably, suggesting that rhamnogalacturonan I was the one most likely synthesized. In hemicellulose fraction, growth regulators reduced arabinose level, suggesting that arabinogalactan II was degraded. And nitrogen source reduced galactose level, suggesting that xyloglucan was also degraded.

• 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.

• Korean Journal of Plant Tissue Culture
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• v.22 no.2
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• pp.115-120
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• 1995

A method for cryopreservation of suspension cultured embryogenic cells derived from immature zygotic embryos of rice (Korean cultivars, Donggin-byeo and Taebaeg-byeo) was developed. The highest cell regrowth after storage in liquid nitrogen was obtained when Donggin-byeo cells were cryoprotected with a mixture of 2 M DMSO and 0.4 M sucrose and Taebaeg-byeo cells with a mixture of 0.64 M DMSO and 0.4 M sucrose at frequencies of 88% and 90%, respectively, Pretreatment in a high osmotic medium was not necessary. Upon transfer to $N_{6}$ medium suplemented with lmg/L NAA and 5 mg/L kinetin, the regenerated calli gave rise to numerous somatic embryos which subsequently underwent development into plantlets. Among approximately 100 plantlets, 25% of them were albinos.s.

• Korean Journal of Plant Tissue Culture
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• v.27 no.3
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• pp.227-232
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• 2000

This study was carried out to elucidate the effect of cadmium on somatic embryogenesis and plant regeneration from cultured cells of Daucus carota L. Embryogenic calli were induced from cotyledon explants of carrot seedlings cultured on MS solid medium supplemente with 1 mg/L 2,4-D Embryogenic cells proliferated on medium supplemented with 1 mg/L 2,4-D were also cultured in liquid MS medium containing various concentrations (50, 100, 200, 500, 1000 $\mu$M) of cadmium for one week and then transferred to MS basal medium. Somatic embryogenesis occurred in suspension culture treated with 50 $\mu$M and 100 $\mu$M cadmium or untreated with cadmium. When cadmium was treated in suspension culture, production of two and four cotyledonary somatic embryos was reduced, but that of three cotyledonary somatic embryo was increased. Two cotyledonary embryos showed higher regeneration frequency than abnormal somatic embryo with one, three and four cotyledon. Regardless of cotyledonary variation, germination frequency of somatic embryos treated with cadmium was decreased in compared with that of embryos in basal medium.

• Korean Journal of Plant Tissue Culture
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• v.26 no.3
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• pp.183-187
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• 1999

To establish an efficient screening system for new herbicides using plant cultured cells, responses of tobacco photomixotrophic cultured (PH) cells to various herbicides with different modes of action were surveyed by measuring the cell growth and ion conductivity in medium. The cells were cultured in Murashige and Skoog (MS) medium containing 0.7mg/L 2,4-D, 0.3mg/L kinetin and 30 g/L sucrose at $25^{\circ}C$ in the light (100 rpm). Chemicals were treated to suspension cultures of tobacco PH cells at the time of subculture. The cell growth and ion conductivity in the medium were investigated on 12 days after chemical treatment. The ion conductivity assay gave well correlated results to the cell growth inhibition data. The responses of tobacco PM cells were dependent on the modes of action of chemicals tested. Atrazine, an inhibitor of photosynthetic electron transport (PET), strongly inhibited both the cell membrane and cell growth ($IC_{50}$/, about 1 $\mu$M). Butachlor (an inhibitor of cell division), glufosinate (an inhibitor of amino acid biosynthesis), and fluridone (an inhibitor of carotenoid biosynthesis) showed a dose-dependent inhibition. However, Quinclorac, a herbicide with an auxin activity, did not affect the cell growth and ion leakage. These results suggested that tobacco PM cells is suitable materials for the simple screening of new herbicides such as PET, amino acid biosynthesis, ceil division inhibitors by measuring the cell growth and ion conductivity.

• Korean Journal of Animal Reproduction
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• v.18 no.4
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• pp.235-244
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• 1995

The present study was designed to demonstrate that ES cell lines efficiently could be isolated from explanted blastocysts of hybrid BCF1 mouse when grown on STO feeder layer derived from mouse fibroblasts in culture medium supplemented with leukemia inhibitory factor (LIF). The expanded blastocysts were attached to mitomycin C-inactivated STO feeder layer and were cultured for 4 days. Four days later the ICM was disaggregated by a short term trypsin treatment (0.25% trypsin / 0.04% EDT A for 2-3 min). The resulting cell suspension was seeded on a new STO feeder layer and covered with DMEM supplemented with 10% FCS, 0.1 mM nonessential amino acid, 0.1 mM sodium pyruvate, 0.1 mM mercaptoethanol and 1,000 U/ml LIF. Colonies of ES-like cells were observed after the second passage. These colonies were repeatedly passaged at approximately 5 day intervals. In this study, five ES-like celllines were isolated by directly explanting blastocysts, but three lines were lost after the 5th passage, possibly due to toxic effects of a new FCS batch. The characterization of developmental potential of isolated cell lines was performed with respect to in vitro differentiation and specific activity of alkaline phosphatase (AP). When cells were cultured in suspension, the aggregates of cell lines were capable of forming simple embryoid bodies (EB), and showed the capacity for forming cystic multilayer EBs. In addition, the cell lines were positive for AP staining, a biochemical marker characteristic of mouse ES cells.