• Archives of Pharmacal Research
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• v.19 no.3
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• pp.219-222
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• 1996

Ephedra olistachya cultures have been known to accumulate $rho-coumaroylamino$ acids by elicitor treatment. Based on their chemical structures, the biosynthetic pathway of$rho-coumaroylamino$acids was postulated and phenylalanine ammonia-lyase (PAL), cinnamic acid 4-hydroxylase (4-CH) and p-coumaroyl CoA: D-Ala p-coumaroyltransferase ($rho-CT$) were supposed to be involved in the pathway. The time course inductions of these enzymes were investigated after treatment of yeast extract, yeast-derived mannan glycopeptide and D-Ala. They were detectable at only 4 hours and reached to their maximum level at 9 hours after onset of elicitor treatment. The activities of PAL and 4-CH were almost disappeared within 24 hours, however, that of $rho-CT$was remained up to 48 hours irrespective of the kind of elicitors. $rho-CT$ showed substrate specificity to D-Ala at crude enzyme extract level.

• Journal of Life Science
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• v.8 no.5
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• pp.604-613
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• 1998

The last enzyme of the sesquiterpen phytoalexin capsidiol synthesis in tobacco cell, 5-epi-aristolochene hydro-xylase which convert 5-epi-aristolochene (EAS) to capsidiol, was cloned by a reverse transcription polymerase chain reaction strategy and cDNA library screening. Cloned CYP-B3 contained high probability amino acid matches to known plant cytochrome P450 sequences and open reading frame with the conserved FxxGxRxCxG heme-binding region. Transcripts of CYP-B3 were not detected in control cells, but induced in elicitor-treated cells. Furthermore, CYP-B3 transcripts were induced by fungal extracts and cellulase but not by other stimuli(chilling, heat shock and 2,4-D). Induction of CYP-B3 transcripts by elicitor treatment was not affected by ancymidol and ketoconazole treat-ments suggesting that an inhibition of hydroxylase activity by Cyt P450 inhibitors resulting from post translational processing event.

• Archives of Pharmacal Research
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• v.18 no.5
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• pp.336-339
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• 1995

Ephedra distachya cultures have been known to accumulate two major $\rho-coumaroylamino$ acids (p-coumaroylgicine and $\rho-coumaroylamino$ -D-alanine) by D-Ala treament. When D-Ala was added together with serial concentrations of yeast-derived elicitor, the accumulation of $\rho-coumaroylamino$ -D-Ala $(\rho-CDA)$ was geatly increased in an additive manner. In feeding experiments, $[1-^14C]$-D-Ala was incorporated into $\rho-CDA$at a rate of 2.2% or 2.3% of added radioactivity, indicating that exogenous D-Ala served as a precursor of the conjugate. $[1-^14C]$-L-Ala wasalso incorporated into p-CDA (0.23%) in the elicitor treated cultures. This fact suggested that at least a part of $(\rho-CDA)$ was produced from active conversion of L-Ala by the elecitation. In order to investigate a possible role of D-Ala as an elicitor of $\rpo-coumaroylamino$ acids $(\rpo-CAA)$, cold D-Ala was added together with labeled L-Ala. Although L-Ala seemed to be incorporated into $\rho-CDA$ by this treatment, the incorporation ratio was too small (0.054%) to draw a clear conclusion. However, the amount of $\rpo-coumaroylglycine$, which did not use D-Ala as a substrate, was also sightly increased by D-Ala treatment irrespective of the presence of elicitor, suggesting that exogenous D-Ala might act as an elicitor of$\rpo-CAA$ as well as a precusor substrate of $\rho-CDA$.

• KSBB Journal
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• v.13 no.5
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• pp.569-576
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• 1998

The effects of carbohydrates, hormones and elicitors on both cell growth and betaine production were investigated in the cell cultures of Lycium chinense Mill. The maximum effect of glucose and sucrose was observed in cells cultured in the presence of 3% and 7% for cell growth and betaine production, respectively. The effect of hormones on cell growth and betaine production was prominent in the presence of 10 ${\mu}$M 2, 4-D, 10 ${\mu}$M NAA and 2.5 ${\mu}$M IAA, whereas cell growth and betaine production were excellent at 2.5 ${\mu}$M BA and 10 ${\mu}$M BA, respectively. Abiotic elicitors such as KCI, MnCl2 and NaCl exhibited an inhibitory role on cell growth in all treatment groups. Betaine production was increased according to increase of concentration of abiotic elicitors. methanol-soluble and insoluble components as biotic elicitor remarkably inhibited cell growth from 2 mg and 6 mg, respectively. Betaine production was increased maximally at 2 mg of biotic elicitors. When growth medium was switched to production medium at two stage culture, it resulted that cell fresh weight and dry weight decreased but betaine content increased about 2.2-fold.

• Korean Journal of Plant Resources
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• v.21 no.2
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• pp.139-143
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• 2008

Enzyme activity and expression of cytochrome P450 gene involved in the pathway of capsidiol biosynthesis were compared in five different solanaceae plants such as red pepper, green pepper, tobacco, potato and egg plant. Base on genomic DNA and/or RT-PCR results, four solanaceae plants such as red pepper, green pepper, tobacco and egg plant possess P450 gene in the genome and specifically expressed by elicitor treatment. However, potato was appeared to have neither P450 nor cyclase gene in the genome. P450 genes did not show any expression in the plants under normal condition, but showed highly specific expression under elicitation condition in various organs and tissue such as leaf, root, stem and culture cells.

• Korean Journal of Medicinal Crop Science
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• v.12 no.5
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• pp.378-383
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• 2004

Scopolamine and hyoscyamine are important anticholinergic compounds. To increase the productivity, we have selected various elicitors and developed culture system using a bubble column bioreactor (BCB). As the same manner of elicitation in flask cultures, the elicitors were introduced into BCB cultures and the productivity was investigated. Except the bacterial elicitor of Staphyllococcus aureus, the elicitors inhibited hyoscyamine production. In scopolamine production, the elicitors revealed different responses from the results obtained in flask cultures. The elicitors of KCl and Candida albicans less increased the production than flask cultures. However, methyl jasmonate and S. aureus showed stronger positive effects on tropane alkaloid production. In particular, S. aureus was the most effective elicitor on scopolamine production and the elicitor resulted in the highly increased production, approximately 10 times higher than the control culture.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.1
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• pp.73-77
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• 2005

Plant-derived natural products have been and will continue to be valuable sources. Elicitors have been employed to modify cell metabolism in order to enhance the productivity of useful metabolites in plant cell/tissue cultures. In this study, several elicitors were used to improve the productivity of useful metabolites and to reduce culture time for archiving high concentration in P. ginseng hairy root cultures. The addition of chitosan, chitosan oligosaccharide and alginate oligosaccharide to the culture of P. ginseng hairy roots caused growth to be inhibited with the increase in elicitor concentration. The usage of the chitosan elicitor and D-glucosamine caused a slight decrease in hairy root growth, whereas total ginseng saponin accumulated slightly with the increase in elicitor concentration. When gel beads were added to the culture medium at the initial period, hairy root growth was enhanced. The maximum growth was 1.35 times higher than that of the control at $1\%$ (w/v). Total ginseng saponin content decreased due to the addition of alginate beads. This would result in consistent diffusion of lower levels of calcium ions during the culture period that promotes biomass growth.

• Korean Journal of Plant Tissue Culture
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• v.21 no.6
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• pp.357-362
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• 1994

Chitinases and $\beta$-1,3-glucanases are believed to be important in defending plane against pathogens. Here, we investigated the expression of chitinase(s) in Arabidopsis thaliana cell suspension culture system in response to ethephon (2-chloroethyl phosphonic acid) which produces ethylene or a microbial elicitor, a bacterial pectin-degrading enzyme, $\beta$-1, 4-endopolygalactronic acid Iyase (PGA Iyase), treatment. Chitinase activity was measured either by radio chemical assay using $^3$H-labeled regenerated chitin as substrate or western blot analysis using antibody raised against tobacro chitinase(S). With 1 mg/mL of ethephon or 100 m units/mL of elicitor treatment, maximum levels of activity were reached after 48h. We also investigated distribution of chitinase activity in seedlings, leaves, and root of A. thaliana and found that root have the highest chitinase activity.

• Archives of Pharmacal Research
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• v.17 no.1
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• pp.26-30
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• 1994

Some ammonium oxalate soluble pectic fragments prepared from cultured cell wall of Ephycla distrahya elicited the accumulation of p-coumarocylamino acids (p-CAA) in E. distachya cultures while water soluble and alkali soluble fractions had no activity. Partial purification of the pectic fragments fraction using DEAE-cellulose chromatography afforded two active fractions (PS-I and PS-II) which were composed of mainly uronic acids (98-99 w/w %). They elicited the accumulation of p-CAA in an amount of 52-60 nmol per gram fresh weight of cultures. The acidic sugar compositions of PS-I and PS-II were found to be galacturonic acid and glucuronic acid by TLC analysis. They were supposed to act as endogenous elicitors of p-CAA accumulation. In order to investigate the effect of ethylene on p-CAA accumulation, Ethrel, which is known as ethylene generator, and ACC(1-aminocyclopropane-1-carboxylic acid), a direct precusor of ethylene biosynthesis, were added to the culture. However, they did not glycopeptide elicitor [(Con A-II)], either. Consequently, no relationships between ethylene and p-CAA accumulation were recognized. Several tentative elicitors were teted for their activity. Commercial yeast glucan, $CuCl_2$, laminarin and laminariheptaose had slight activity whereas ${\alpha}$-methylmannopyranoside and commercial yeast mannan had no elicitor activity. ${\alpha}$-methylmannopyranoside which has been known as a tentative inhibitor of glucan elicitor in Glycine max did not affect on the elicitor activity of Con A-II.

• Journal of Microbiology and Biotechnology
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• v.9 no.5
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• pp.661-667
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• 1999

The effect of fungal elicitor and heavy metal salts on the production of flavonol glycosides in cell cultures of Ginkgo biloba was investigated. Among the fungi tested, Trichoderma longibrachiatum ATCC 52326 was found to be the most efficient in the production of flavonol glycosides. Kaempferol production from the elicited callus increased ten-fold as compared to the unelicited callus, while quercetin concentration of elicited cells was nine-fold higher than that of uneliceited cells in suspension cultures. The maximum quercetin concentration of 0.362㎎/l was obtained in 1.25㎎/l of the homogenate elicitor. Among the heavy metal salts tested, CuSO₄ showed a significant effect on quercetin accumulation, reaching to the concentration of 0.526 ㎎/l. Quercetin concentration increased to a maximum of l2-fold in response to CuSO₄ treatment as compared to that of untreated cells. The phenylalanine ammonia-lyase (PAL) activity and flavonol glycosides production simultaneously increased for 5 days of culture after fungal elicitor feeding, and their contents showed the same proportional patterns during the culture period. In contrast, PAL activity of cell cultures treated with CuSO₄ was almost constant during the culture period, although quercetin production increased remarkably.