• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.78.1-78
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• 2003

An osmotin-like protein (CAOSMl) gene was isolated from pepper leaves infected with the avirulent strain Bv5-4a of Xmthomonas campestris pv. vesicatoria. The cDNA encodes a polypeptide of 250 amino acids with a molecular mass of 27, 361 Da. Its amino acid sequence is highly homologous to various osmotin-like proteins from other plant species. The CAOSMl gene expression was organ- and tissue-specifically regulated In pepper plants. The CAOSMl mRNA was intensely localized in the endodermis area of root tissue and in the phloem cells of vascular bundles of red fruit tissue, but not in leaf, stem, and green fruit tissues of healthy pepper plants. Infection by X. c. pv vesintoria, Colletotrichum coccodes, or Phytopkhora capsici iinduced CAOSMl transcription in the leaf or stem tissues. Expression of the CAOSMl gene was somewhat higher in the incompatible than the compatible interactions of pathogens with pepper. The CAOSMl mRNA was prevalently localized in the phloem cells of the vascular bundle of leaf tissues infected by C. coccodes. The CAOSMl gene was activated in leaf tissues by treatment with ethylene, methyl jasmonate, high salinity, cold acclimation and mechanical wounding, but not by abscisic acid (ABA) and drought. These results indicate that the pepper CAOSMl protein functions in response to Pathogens and some abiotic stresses in pepper plants

• Korean Journal of Medicinal Crop Science
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• v.15 no.4
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• pp.241-245
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• 2007

Effects of various concentrations of elicitors for enhancement in valepotriates and valerenic acids production in adventitious root culture of Valeriana fauriei were investigated. Although the growth of adventitious roots was suppresed by addition of biotic or abiotic elicitors, the production of valepotriates and valerenic acids was enhanced. Addition of $100\;{\mu}M$ methyl jasmonate or 1 $g/{\ell}$ yeast extract to the root culture of V. fauriei resulted in the optimal production of valepotriates ($12.56\;{\pm}\;0.78\;mg/{\ell}$) and valerenic acids ($10.63\;{\pm}\;1.1\;mg/{\ell}$), respectively.

• Horticultural Science & Technology
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• v.31 no.3
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• pp.299-307
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• 2013

A radish (Raphanus sativus L.) polygalacturonase-inhibiting protein (PGIP) gene was cloned and compared to the PGIP gene (BrPGIP2) from Chinese cabbage (Brassica rapa ssp. pekinensis) in order to gain more information on controlling a disease and improving produce quality. To clone the radish PGIP gene, primers were designed based on conserved sequences of two PGIP genes (BnPGIP1 and BnPGIP2) from rape (B. napus L. ssp. oleifera), Chinese cabbage and Arabidopsis thaliana. PCR cloning was performed with cDNA from the stigma of radish 'Daejinyeoreum' as a template to confirm DNA fragments which were about 600 base pair in size. Sequence analysis revealed 84.1% homology with BrPGIP2 and 70.1% with BnPGIP1. DNA walking was conducted to confirm the open reading frame of 972 bp, and the gene was named RsPGIP1. RsPGIP1 consisting with 323 amino acids (aa) has a high leucine content (54/323) and contains 10 leucine-rich repeat domains, as do most BrPGIPs of Chinese cabbage. The gene expression of RsPGIP1 was induced by abiotic stresses and methyl jasmonate. It showed enrichment in the stigma and the primary root than a leaf. Cloning RsPGIP1 will aid to further apply practices on postharvest quality maintenance and disease control of the root.

• Journal of Plant Biotechnology
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• v.2 no.2
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• pp.79-82
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• 2000

We have isolated a cDNA encoding a calcium-dependent protein kinase (CDPK) in Nicotiana tabacum, which was designated NtCDPK1. Accumulation of the NtCDPK1 mRNA was stimulated by various stimuli, including phytohormones, CaCl$_2$ wounding, fungal elicitors, chitin and methyl jasmonate. The NtCDPK1 gene encodes a functional Ser/Thr protein kinase of which phosphorylation activity is strongly induced by calcium. By analyzing expression of the NtCDPK1-GFP fusion protein and by immunoblotting with antibody which reacts with NtCDPK1, we found that NtCDPK1 is localized in membrane and nucleus in plant cells. Silencing expression of the NtCDPK1 transgene resulted in marked decrease of lateral root development in the transgenic tobacco plants. Yeast two hybrid screening using NtCDPK1 as a bait identified a tobacco homologue of proteasome regulatory subunit 21D7, designated Nt21D7. The 21D7 mRNA has been shown to be predominantly expressed in proliferating tissues in the cell cycledependent manner in carrot. The recombinant NtCDPK1 protein associated with Nt21D7 in vitro, and could phosphorylate the Nt21D7 protein in vitro in the presence of calcium, suggesting that Nt21D7 protein is a natural substrate of NtCDPK1 in tobacco. These results suggest that NtCDPK1 may regulate tell proliferation processes, such as lateral root formation, by regulating specificity and/or activity of proteasome-mediated protein degradation pathway.

• Plant Biotechnology Reports
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• v.4 no.2
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• pp.165-172
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• 2010

Genes that are expressed early in specific response to high salinity conditions were isolated from rapeseed plant (Brassica napus L.) using an mRNA differential display method. Five PCR fragments (DD1.5) were isolated that were induced by, but showed different response kinetics to, 200 mM NaCl. Nucleotide sequence analysis and homology search revealed that the deduced amino sequences of three of the five cDNA fragments showed considerable similarity to those of ${\beta}$-mannosidase (DD1), tomato Pti-6 proteins (DD5), and the tobacco harpin-induced protein hin1 (DD4), respectively. In contrast, the remaining clones, DD3 and DD2, did not correspond to any substantial existing annotation. Using the DD3 fragment as a probe, we isolated a full-length cDNA clone from the cDNA library, which we termed BnSWD1 (Brassica napus salt responsive WD40 1). The predicted amino-acid sequence of BnSWD1 contains eight WD40 repeats and is conserved in all eukaryotes. Notably, the BnSWD1 gene is expressed at high levels under salt-stress conditions. Furthermore, we found that BnSWD1 was upregulated after treatment with abscisic acid, salicylic acid, and methyl jasmonate. Our study suggests that BnSWD1, which is a novel WD40 repeat-containing protein, has a function in salt-stress responses in plants, possibly via abscisic acid-dependent and/or -independent signaling pathways.

• Korean Journal of Medicinal Crop Science
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• v.25 no.2
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• pp.95-101
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• 2017

Background: Callus cultivation has the advantage of producing a large amount of tissue of a plant in a laboratory regardless of the environment, for extracting an active substance. In the present study, callus formation was induced in the leaves of the succulent plant Adenium obesum (Forssk.) Roem & Schult. After callus cultivation, anti-inflammatory activity tests were conducted, because leaves and stems of A. obesum have been reported to possess biological activity. Methods and Results: In order to induce callus formation, various concentrations of plant growth factors, such as kinetin, naphtha-leneacetic acid (NAA), 6-benzyladenine (BA), and indole-3-acetic acid (IAA) were added to MS solid medium. The maximum callus proliferation was induced by mixed medium consisting of NAA ($2mg/{\ell}$) and BA ($1mg/{\ell}$). In addition, an elicitor was added to the medium under optimal conditions for initiating suspension culture. After suspension culturing, the activities of the callus extracts were compared and analyzed. The cytotoxicity and anti-inflammatory activity tests revealed that the anti-inflammatory activity of the callus extract and the content of phenolic compounds were elevated after treatment of the callus culture with the elicitior. Conclusions: A. obesum callus might be considered as potential source of biologically active anti-inflammatory material.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.61 no.1
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• pp.33-40
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• 2016

This study is focused on the evaluation of growth parameters, total polyphenol content (TPC), chlorophyll content as well as the DPPH (1,1-diphenyl-2-picryhydrazyl) free radical scavenging activity of young barley seedling (YBS) affected by elicitation. Salicylic acid (SA), methyl jasmonate (MJ), amino acid liquid fertilizer (ALF) and microbial metabolism activator (MMA) were used. Elicitation was conducted for two times and various concentrations were used in this study. The result revealed that, MJ 1 ml/L treated-YBS gave the longest seedling length of 1.33 cm, followed by the ones treated with SA 1.38 mg/L and ALF 2 ml/L, respectively. ALF 3 ml/L treatment gave the highest fresh weight of 10 seedlings, followed by MJ 5 ml/L and SA 13.8 mg/L treatment with 1.56 g, 1.55 g and 1.53 g respectively. SA 138.12 mg/L elicitor treated-YBS gave the highest Chl a, Chl b content of $8.57{\mu}g/mg$ and $3.83{\mu}g/mg$, respectively while the highest carotenoid content was found in MJ ml/L treatment with $1.62{\mu}g/mg$. Among elicitor treated-YBS, SA showed better TPC. The highest TPC was found in SA 1.38 mg/L treatment with 18.82 mg/g TAE. Likewise, SA 1.38 mg/L showed the highest DPPH free radical scavenging activity among all the treatments. However, the lowest TPC was found in ALF 1ml/L treated-YBS with 9.46 mg/g TAE, which was even lower than the control (14.31 mg/g TAE).

• Molecules and Cells
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• v.19 no.1
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• pp.67-73
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• 2005

Isoflavones are synthesized by isoflavone synthases via the phenylpropanoid pathway in legumes. We have cloned two isoflavone synthase genes, IFS1 and IFS2, from a total of 18 soybean cultivars. The amino acid residues of the proteins that differed between cultivars were dispersed over the entire coding region. However, amino acid sequence variation did not occur in conserved domains such as the ERR triad region, except that one conserved amino acid was changed in the IFS2 protein of the GS12 cultivar ($R_{374}G$) and the IFS1 proteins of the 99M06 and Soja99s65 cultivars ($A_{109}T$, $F_{105}I$). In three cultivars (99M06, 99M116, and Simheukpi), most of amino acid changes were such that the difference between the amino acid sequences of IFS1 and IFS2 was reduced. The expression profiles of three enzymes that convert naringenin to the isoflavone, genistein, chalcone isomerase (CHI), isoflavone synthase (IFS) and flavanone 3-hydroxylase (F3H) were examined. In general, IFS mRNA was more abundant in etiolated seedlings than mature plants whereas the levels of CHI and F3H mRNAs were similar in the two stages. During seed development, IFS was expressed a little later than CHI and F3H but expression of these three genes was barely detectable, if at all, during later seed hardening. In addition, we found that the levels of CHI, F3H, and IFS mRNAs were under circadian control. We also showed that IFS was induced by wounding and by application of methyl jasmonate to etiolated soybean seedlings.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.2
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• pp.155-161
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• 2005

The inherent instability of metabolite production in plant cell culture-based bioprocessing is a major problem hindering its commercialization. To understand the extent and causes of this instability, this study was aimed at understanding the variability of anthocyanin accumulation during long-term subcultures, as well as within subculture batches, in Vitis vinifera cell cultures. Therefore, four cell line suspensions of Vitis vinifera L. var. Gamay Freaux, A, B, C and D, originated from the same callus by cell-aggregate cloning, were established with starting anthocyanin contents of $2.73\;\pm\;0.15,\;1.45\;\pm\;0.04,\;0.7\;\pm\;0.024\;and\;0.27\;\pm\;0.04$CV (Color Value)/g-FCW (fresh cell weight), respectively. During weekly subculturing of 33 batches over 8 months, the anthocyanin biosynthetic capacity was gradually lost at various rates, for all four cell lines, regardless of the significant difference in the starting anthocyanin content. Contrary to this general trend, a significant fluctuation in the anthocyanin content was observed, but with an irregular cyclic pattern. The variabilities in the anthocyanin content between the subcultures for the 33 batches, as represented by the variation coefficient (VC), were 58, 57, 54, and $84\%$ for V. vinifera cell lines A, B, C and D, respectively. Within one subculture, the VCs from 12 replicate flasks for each of 12 independent subcultures were averaged, and found to be $9.7\%$, ranging from 4 to $17\%$. High- and low-producing cell lines, VV05 and VV06, with 1.8-fold differences in their basal anthocyanin contents, exhibited different inducibilities to L-phenylalanine feeding, methyl jasmonate and light irradiation. The low-producing cell line showed greater potential in enhanced the anthocyanin production.

• Journal of Plant Biotechnology
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• v.5 no.2
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• pp.121-129
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• 2003

Callus cultures of Azadirachta indica flower petals were established on MS medium supplemented with naphthalene acetic acid (1 mg/L), kinetin (0.5 mg/L) and sucrose $(3\%\;w/v)$. Cell cultures of Azadirachta indica were established and studied the growth and production kinetics. Half 85 medium supplemented with dicamba (2 mg/L), kinetin (1 mg/L) and sucrose $(3\%\;w/v)$ was found to be suitable for initiation and maintenance of cell cultures from the calli. MS medium supplemented with naphthalene acetic acid (NAA) (1 mg/L), kinetin (0.5 mg/L) and sucrose $(3\%\;w/v)$ was found to be suitable as production medium. Around $80\%\;(0.05\%\;w/v)$ of azadirachtin was found to be intracellular. The effect of various precursors, elicitors, permeabilizing agents and growth retardants in cell cultures was studied. The addition of precursors sodium acetate (10 mg/L), squalene (10 mg/L), isopentenyl pyrophosphate (1 mg/L) and geranyl pyrophosphate (1 mg/L) to the cell cultures on day 3 has shown significant increase in bioproduction of azadirachtin $(64.94{\pm}4.40\;mg/L,\;72.81{\pm}0.04\;mg/L,\;51.63{\pm}1.26\;mg/L\;and\;30.70{\pm}0.28\;mg/L\;respectively)$ over the control cultures $(4.70{\pm}0.27 mg/L)$. $5\%$ v/v cell extracts of Fusarium solani has shown moderate increase in the content of azadirachtin $(5.71{\pm}0.34\;mg/L)$ when compared to control cultures $(2.40{\pm}0.56\;mg/L)$. The addition of methyl jasmonate $(500\;{\mu}M/L)$ on day 3 has shown $\~4$ fold improvement in bioproduction of azadirachtln $(6.92{\pm}0.11\;mg/L)$ when compared to control cultures $(1.63{\pm}0.02\;mg/L)$. There was no significant effect of the studied growth retardants and permeabilizing agents on bioproduction of azadirachtin. Cells are cultivated in large volumes using the effective precursors.