• The Plant Pathology Journal
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• v.26 no.4
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• pp.386-393
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• 2010

In order to study disease resistance mechanisms in rice against the rice blast fungus Magnaporthe grisea, we screened fungal elicitor-responsive genes from rice suspension-cultured cells treated with fungal elicitors employing differential hybridization (DH). By DH screening, 31 distinct rice clones were isolated and a majority of them were full-length cDNAs encoding pathogenesisrelated (PR) genes. Sixteen of the 31 genes were upregulated at 4, 8, and 12 h following fungal elicitor treatment. To elucidate the effect of signal molecules and biotic elicitors on the regulation of rice defense genes, we further characterized the transcriptional expression patterns of representative isolated PR genes; OsGlu1, OsGlu2, OsTLP, OsRLK, and OsPR-10, following treatment with fungal elicitor, phytohormones, cycloheximide, and inhibitors of protein phosphorylation. Jasmonic acid (JA) induced transcriptional expression of OsGlu1, OsTLP, and OsRLK, but not of OsGlu2 and OsPR-10 at any of the tested time points. Salicylic acid (SA) and abscisic acid weakly induced the expression of OsTLP and OsRLK. SA showed an antagonistic effect with fungal elicitor and JA. Cycloheximide suppressed all these genes upon elicitor treatment, except for OsGlu2. Staurosporine only induced the expression of OsRLK. Application of calyculin A strongly induced OsRLK expression, but suppressed the expression of OsGlu2. Our study yielded a number of PR genes that play a role in defense mechanisms against the rice blast fungus, as well as contribute towards the elucidation of crosstalk between phytohormones and other modifications during defense signaling.

• Korean Journal of Plant Tissue Culture
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• v.22 no.3
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• pp.157-164
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• 1995

Intact mature zygotic embryos or their excised cotyledons of ginseng, were cultured on media containing various growth regulators such as auxin (2,4D, IAA) and cytokinin(BAP kinetin). In the culture of intact zygotic embryos, auxin inhibited germination but cytokinin did not Somatic embryogenesis occurred only from those of ungerminated embryos. In the culture of cotyledon segment, medium without growth regulators was the most appropriate to somatic embryogenesis. Somatic embryos were produced sporadically over the surfaces of zygotic embryos on medium containing auxin, while on medium without growth regulators, or media containing cytokinin, somatic embryos formed only on the proximal region of cotyledon. on medium containing 2,4-D, somatic embryos originated from multiple cells which comprised epidermal and subepidermal layers of cotyledon, which resulted in poly-somatic embryogenesis. When these somatic embryos were cultured on the same medium, the primary somatic embryos procured secondary embryos, which arose from epidermal or subepidermal single cells.

• Korean Journal of Plant Resources
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• v.19 no.6
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• pp.706-715
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• 2006

Nitrate reductase deficient (NR) mutant lines were selected indirectly by their resistance to 100mM chlorate in cell cultures of P. parviflora. A total of 585 chlorate resistant lines were confirmed by a second passage on a high concentration of chlorate. Frequency of spontaneous mutation was $9.7{\times}10^{-7}$ in 3 month old suspension-cultured cells, and in non-selective media containing amino acids as sole nitrogen source. The frequency of mutation could be increased up to 11-fold by culture for 12 months. Out of 40 randomly selected calli, 22 were fully deficient in NR. The rest of the clones contained a decreased level of NR activity. Further characterization was carried out in 13 mutant lines which were fully deficient in NR and in 5 mutant lines containing residual (0-7.0%) NR activity, as compared to wild-type cells cultured on the same medium. The $NR^-$ mutants were tentatively classified as defective in the NR apoenzyme (nia-type; 11 mutant lines including the 5 with residual NR activity) or in the molybdenum cofactor (cnx-type; 7 mutant lines) by the XDH activity. The cnx-type could be further classified into two groups. In one group (5 mutant lines) of these, the NR activity could be partially restored by nonphysiologically high (1.0mM) molybdate in the culture medium. Both types of $NR^-$ mutants were unable to grow on minimal medium containing nitrate as sole nitrogen source, but grew well on amino acids. They also proved to be extremely sensitive to the standard medium ($MSP_1$) containing nitrate and ammonium. Shoot regeneration was obtained only in the $NR^-$ mutants, which contained residual NR activity, but they so far have failed to grow into plants.

• Korean Journal of Plant Resources
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• v.27 no.6
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• pp.707-713
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• 2014

This study examined the inflammatory reaction effects of Allium victorialis var. platyphyllum in vivo at the time of a lipopolysaccharide (LPS) shock in rats, and in vitro in cultured Raw 264.7 cells, with the aim of facilitating the development of a new anti-inflammatory medicine. Plasma concentrations of interleukin (IL)-$1{\beta}$, IL-6, tumor necrosis factor ${\alpha}$ (TNF-${\alpha}$), and IL-10 in rats peaked 5 h after LPS treatment in all experimental groups, with those of IL-$1{\beta}$, IL-6, and TNF-${\alpha}$ being significantly lower in all animals treated with A. victorialis than in the control group at that time point. Conversely, the plasma concentration of IL-10 was higher in the rats treated with 300 mg/kg A. victorialis extract than in the control group at both 2 and 5 h after LPS treatment. Concentrations of IL-$1{\beta}$ and IL-6 in the liver of rats treated with A. victorialis extract were significantly lower than those of the saline-treated control group. However, the liver concentrations of TNF-${\alpha}$ and IL-10 did not vary significantly between the four animal groups. Similarly, concentrations of IL-$1{\beta}$, IL-6, and TNF-${\alpha}$ obtained from cultured Raw 264.7 macrophages were lower in all of the A.-victorialis-extract-treated groups than in the control group. Although the concentration of IL-10 in the A.-victorialis-extract-treated groups tended to be greater than in the control group, the differences between groups were not statistically significant. Together the findings of this study suggest that A. victorialis var. platyphyllum contains functional substances that are involved in inflammatory reactions.

• Natural Product Sciences
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• v.18 no.3
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• pp.177-182
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• 2012

Oxidative stress potentially induces neurotoxicity which is believed to underlie several major age-related diseases of the central nervous system. This study sought to identify the cytoprotective effects of sixty-nine Cambodian plants against glutamate-induced cell death. Cultured HT22 cells were applied as an in vitro model, and neurotoxicity was induced in these neuronal cells by exposure to a determined concentration of glutamate. Sixty-nine plant sources, as Cambodia's indigenous species, were purchased from O'reusey Market, Phnom Penh, and extracted with ethanol. These extracts were screened for cytoprotective effects against glutamate-triggered neurotoxicity in HT22 cells at concentrations of 100 and 300 ${\mu}g/ml$. Of these, eight ethanol extracts, bark of Anacardium occidentale, bark and sapwood of Bauhinia pulla, flowers of Borassus flabellifer, stems and leaves of Coix lacryma-jobi, bark and sapwood of Diospyros nitida, sapwood of Dipterocarpus obtusifolius, stems of Oryza rufipogon, and fruits of Phyllanthus emblica, showed significant cytoprotective effects against glutamate-induced cell damage and degeneration in HT22 cells.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.14 no.2
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• pp.79-85
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• 1981

As one of the fundamental studies for the breeding of marine algae, the effects of several plant hormones (IAA, Gibberellin, 2.4-D, NAA, Kinetin) on the growth of Porphyra-fronds, P. tenera Kjell. form tamatsuensis Miura, were investigated from January 21 to February 21 1981. The fronds used for the experiment were dissected out at $25mm^2$ size, and cultured in modified Provasoli's ESP medium supplemented with various concentrations of each plant growth regulators. The culture was kept under constant water temperature of $5^{\circ}C$ in 14 hrs. photoperiod and illuminated with 2,400 lux by fluorescent light. Based on the results of first experiment, the culture of fronds for the secondary experiment was carried out at $5^{\circ}C\;and\;10^{\circ}C$ in medium containing various levels of Kinetin from April 6 to 24, and compared the growth of two groups at each concentrations with each other, The results obtained are summarized as follows : (1) The best growth efficiencies were observed at 5.0mg/1 of each plant hormones except Gibberellin. Among them, the highest growth-rate was $312.5\%\;(345.3\%\;in\;frond\;size)$ in contrast with control at 5.0mg/1 of Kinetin, and was followed by $257.5\%\;(236.1\%)$ in 2.4-D,$166.7\%(147.6\%)$ in IAA and $141.7\%\;(167.7\%)$ in NAA, but that in Gibberellin was $247.9\%(241.9\%)$ at 10.0mg/l. (2) Especially, the fronds cultured at 5.0mg/1 of Kinetin were deep black-brown in colour, and had vivid, healthy chloroplasts in their all cells. On the contrary, the fronds cultured in other media were discoloured to light black-brown or green-drown, and almost all cells were vacuolated or shrunk gradually into death.(3) There was an obvious difference between the best growth-rates of the fronds cultured at 5.0mg/l of Kinetin at $5^{\circ}C$ and those at $10^{\circ}C$. The former was $366.7\%$, the latter $318.8\%$ but the difference was little at lower concentrations. (4) Many abnormal cells grown up to $25.0-27.5\mu$ in diameter were found among the marginal cells of fronds which showed the best growth in Kinetin, and the fronds wire $41.0-42.0\mu$ in thickness which was thicker by $10.0\mu$ or so than the others. (5) In two fronds at 1.0mg/1 of Kinetin cell-divisions were observed, which might developed into antheridium, but it was doubtful whether due to the efficiency of Kinetin.

• Korean Journal of Plant Tissue Culture
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• v.21 no.1
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• pp.41-46
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• 1994

Calli were induced from leaf explants of B.falcatum, and selected cell clumps of the calli (900-1, 000${\mu}{\textrm}{m}$) were cultured on MS medium supplemented with 0.1, 0.5, 1.0 or 2.0 mg/L 2, 4-D for 7 days, respectively: The clumps were subsequently transferred onto MS basal medium and subcultured for four weeks. In order to investigate the effect of 2, 4-D pretreatment, the selected clumps were cultured on MS medium supplemented with 0.1 mg/L 2, 4-D for 24, 48, 72, 96, 120 or 144 hours and then transferred to liquid MS basal medium, wherein they were cultured for 4 weeks. Histological observation showed that root initial cells were developed from cells on the surface of clumps or from cells in the inner region. Clumps on the basal medium produced mot within 5 days of culture. The rate of prutruding time was inversely proportional to the concentration of 2, 4-D. The number of adventitious roots per clump preheated with 0.1 mg/L 2, 4-D was an average of 5.2, which was the highest level. On MS medium as control, the clumps formed 3.3 adventitious roots each. As tile concentration of 2, 4-D increased, the number of adventitious roots were declined accordingly: The number of adventitious roots as the period of pretreatment increased upto 120 h.

• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.121-126
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• 1999

Calli were induced from the petiole explants of Pimpinella brachycarpa on MS medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L BA after four weeks of culture. Compact clusters of small and dense cells among these calli were selected and suspension-cultured as the source of embryogenic calli. When transferred to MS medium with 0.1 mg/L NAA, the suspension-cultured cells grew to embryogenic callus. Somatic embryos derived from these embryogenic calli developed into plantlets. The cDNA library was constructed in the embryogenic callus and in order to screen the cDNA library, these cDNAs were plated at a density 1.5 $\times$ 10^5 plaques per 15 cm petridish. Among 19 clones showing preferential hybridization with petiole HD-Zip gene, five clones were obtained after second screening. Four clones among them, were highly homologous to P. brachycarpa shoot-tip Phz4 gene, but one clone, Phc5 was about 1.5 kb which has an extra 163 bp to 5' upstream of Phz4. The Phc5 was 1,531 bp containing poly A tails of 18 bases. ATG start codon for Phc5, was located at position 284 with an open reading frame of 906 by which encodes a polypeptide of 302 amino acids. The Phc5 protein revealed that the polypeptides between 135 and 195 contain a homeodomain as the `leucine zipper' motif.

• Journal of Microbiology and Biotechnology
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• v.6 no.2
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• pp.132-136
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• 1996

To investigate the characteristics of immobilized peppermint (Mentha piperita) cells, dry cell weight (DCW), change of cell viability, and oil productivity of the immobilized cells were determined. Peppermint cells were immobilized in polyurethane (PU) foams of $5{\times}5{\times}5$ mm and cultured in a shaking flask. The maximum DCW was 2.1 mg per foam piece after 20 days of cultivation and the cell density was approximately 420 mg per flask containing 200 foams in 200 ml medium. For the first five days of cultivation, the cell viability was about 80$%$ and decreased to 70$%$ during 5 to 20 days of cultivation. The maximum oil productivity, 148 mg/l was achieved after 40 days of cultivation. The immobilized cells were also cultivated in a bioreactor, equipped with a round spiral type impeller, containing 2, 400 PU foams. The cell viability after 30 days of cultivation with chitosan as an elicitor in the bioreactor was 67$%$ and DCW was 2.0 mg per foam piece. Though the cell viability was relatively high in the bioreactor system, the oil productivity was relatively lower than that of the flask system.

• Journal of Plant Biotechnology
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• v.6 no.1
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• pp.25-37
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• 2004

Successful genetic transformation of plants requires non-chimeric selection of transformed tissues and their subsequent regeneration. With rare exceptions, most transformation protocols still rely heavily on antibiotics for selecting transgenic cells that contain an antibiotic-degrading selectable marker gene. Here, the morphogenic capacity of in-vitro explants of chrysanthemnum and tobacco stems and leaves (control and transgenic) changed with the addition of aminoglycoside antibiotics (AAs), In a test of 6 AAs, phytotoxicity occurred at concentrations of 10 to 25 and 50 to 100$\mu\textrm{g}$ $mL^{-1}$ in chrysanthemum and tobacco explants, respectively. Light conditions as well as explant source and size also had significant effects. The use of transverse thin cell layers (tTCLs), in conjunction with high initial AA selection levels, supported the greatest regeneration of transgenic material (adventitious shoots or callus) and the lowest number of escapes. Flow-cytometric analyses revealed no endodu-plication in chrysanthemum, even at high AA levels. However, this phenomenon was observed in tobacco calli(8C or more), even at low AA concentrations (i.e., 5 to 10 $\mu\textrm{g}$ mL$^{-1}$ ).