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ABSTRACT: It has been recognized that ginsenoside Rg3 is not naturally produced in ginseng although this ginsenoside can accumulate in red ginseng as the result of a thermal process. In order to determine whether or not Rg3 is synthesized in ginseng, hairy roots were treated with methyl jasmonate (MJ). From HPLC analysis, no peak for Rg3 was observed in the controls. However, Rg3 did accumulate in hairy roots that were MJ-treated for 7 days. Rg3 content was 0.42 mg/g (dry weight). To gain more insight into the effects of MJ on UDP-glucosyltransferase (UGT) activity, we attempted to evaluate ginsenoside Rg3 biosynthesis by UGT. A new peak for putative Rg3 was observed, which was confirmed by LC-MS/MS analysis. Our findings indicate that the proteins extracted from our hairy root lines can catalyze Rg3 from Rh2. This suggests that our ginseng hairy root lines possess Rg3 biosynthesis capacity.
Plant Cell Tissue and Organ Culture 01/2013; 112:87-93. · 3.09 Impact Factor
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ABSTRACT: In order to produce centellosides from whole plant cultures of Centella asiatica (L.) Urban, we evaluated the synergistic effects of thidiazuron (TDZ) and methyl jasmonate (MJ) on whole plant growth and In order to produce centellosides from whole plant cultures of Centella asiatica (L.) Urban, we evaluated the synergistic effects of thidiazuron (TDZ) and methyl jasmonate (MJ) on whole plant growth and
centelloside production. After 4weeks of treatment with 0.025mg/L of TDZ coupled with 0.1mM MJ, the production of madecassoside centelloside production. After 4weeks of treatment with 0.025mg/L of TDZ coupled with 0.1mM MJ, the production of madecassoside
and asiaticoside from whole plant cultures was estimated to be 2.40- and 2.44-fold, respectively, above that of MJ elicitation and asiaticoside from whole plant cultures was estimated to be 2.40- and 2.44-fold, respectively, above that of MJ elicitation
alone. When whole plants were treated with a growth regulator and an elicitor, the growth of whole plants, as compared to alone. When whole plants were treated with a growth regulator and an elicitor, the growth of whole plants, as compared to
the controls, did not differ. Additionally, total phytosyterol content in the leaves of whole plants co-treated with MJ and the controls, did not differ. Additionally, total phytosyterol content in the leaves of whole plants co-treated with MJ and
TDZ was 1.08-fold greater than those of MJ alone. These results demonstrate that combined treatments not only stimulate the TDZ was 1.08-fold greater than those of MJ alone. These results demonstrate that combined treatments not only stimulate the
accumulation of centellosides in the leaves but also inhibit the reduction of phytosterol levels caused by MJ elicitation. accumulation of centellosides in the leaves but also inhibit the reduction of phytosterol levels caused by MJ elicitation.
KeywordsAsiaticoside– KeywordsAsiaticoside–
Centella asiatica Centella asiatica
–Thidiazuron–Madecassoside–Methyl jasmonate–Phytosterols –Thidiazuron–Madecassoside–Methyl jasmonate–Phytosterols
Plant Biotechnology Reports 05/2012; 5(3):283-287. · 1.19 Impact Factor
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ABSTRACT: The control of bolting time in onion is an important approach for bulb and seed production, as onion plants which bolt do
not produce marketable bulbs and seed yields are dependent on floral induction. However, genetic and molecular studies about
bolting time in onion plants have not been examined yet to date. In order to understand the regulation of bolting time in
onion plants, we conducted the genetic crosses between late bolting-type cultivar (MOS8) and very early bolting-type cultivar
(Guikum). Segregation ratio of late to very early in F2 populations indicated that this lateness trait was determined by a dominant locus. We also analyzed protein profiles in onion
plants with different bolting time by a proteomics approach. Interestingly, a protein spot with significant similarities to
chromodomains of mammalian chromo-ATPase/helicase-DNA-binding 1 or heterochromatin protein 1, which is involved in the histone
modifications, was identified. Histone methyltransferase activity was also observed in onion plants. Taken together, these
results suggest that a genetic pathway may be involved in the modulation of bolting time in onion plants, though there is
no direct evidence that this protein spot obtained by proteomics is relevant to vernalization.
Journal of Plant Biology 04/2012; 52(6):602-608. · 1.07 Impact Factor
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ABSTRACT: This study describes the identification of Panax species using a peptide nucleic acid (PNA) microarray. P. ginseng, P. quienquefolius, and P. japonicus were distinguished from each other using 5 PNA probes designed based on three single nucleotide polymorphisms (SNPs) detected
in internal transcribed spacer (ITS) and 5.8S rDNA regions. Signal intensity comparison between PNA and DNA microarrays revealed
that the PNA microarray provides a significantly more stable and specific fluorescent signal intensity than the DNA microarray.
Three Panax species identified by the PNA microarray were denoted as barcode numbers depending on their fluorescent signal patterns of
each species using 5 PNA probes (PG-ITS-116, PG-ITS-414-1, PG-ITS-414-2, PG-ITS-425-1, and PG-ITS-425-2). P. ginseng, P. quinquefolius, and P. japonicus were denoted as ‘11010’, ‘00202’ and ‘00000’, respectively. The PNA microarray developed in this study will be useful for
legitimizing the distribution of ginseng in domestic and foreign ginseng markets.
Keywords
Panax ginseng
-Internal transcribed spacer-5.8S ribosomal DNA-Peptide nucleotide acid-Microarray-single nucleotide polymorphism
Genes & genomics 04/2012; 32(5):463-468. · 0.44 Impact Factor
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ABSTRACT: Culture conditions were optimized for somatic embryogenesis ofPanax ginseng. The highest frequency of embryo formation was obtained when tissues were excised from the middle region of the cotyledon
segments of zygotic embryos. Only treatment with light could stimulate the formation of single-type somatic embryos, whereas
multiple-type somatic embryos and calli were observed under dark conditions. The highest production of somatic embryos was
found with an NH4
+:NO3 ratio of 21:39. Among the tested media (MS, B5, and SH), maximum formation of somatic embryos was obtained when cotyledon
expiants were cultured on an 1% agar MS medium supplemented with 5% sucrose. Regenerated ginseng plantlets were transferred
to an autoclaved soil mixture in the greenhouse. These transformants showed no detectable variations in their morphology or
growth characteristics compared with the donor plant.
Keywordsagar-macro salts-
Panax ginseng
-somatic embryogenesis-sucrose
Journal of Plant Biology 04/2012; 49(5):348-352. · 1.07 Impact Factor
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ABSTRACT: In this study, methyl jasmonate (MJ)-induced changes of triterpene saponins in ginseng (Panax ginseng C.A. Meyer) hairy roots and expression profiling of relevant responsive genes were analyzed. The transcription of PgSS (squalene synthase), PgSE (squalene epoxidase), and PNA (dammarenediol synthase-II) genes in hairy root cultures elicited by MJ treatment increased as compared with the controls,
whereas that of PNX (cycloartenol synthase) decreased slightly. In order to select candidate genes encoding for cytochrome P450-dependent hydroxylase
or glucosyltransferase associated with triterpene biosynthesis, RT-PCR analysis was conducted following MJ elicitation. No
differences were observed in any expression among the five genes associated with the cytochrome P450 family, when compared
to that of control. For candidates of the glucosyltransferase gene,expression of EST IDs PG07020C06, PG07025D04, and PG07029G02
was upregulated. In an effort to assess the effects of MJ elicitation on the biosynthesis of triterpene saponin, protopanaxadiol
saponin (Rb group) and protopanaxatriol saponin (Rg group) contents in hairy roots were evaluated by HPLC analysis. With 7
days of MJ elicitation, levels of all ginseonsides of the two-groups increased much higher than that observed in the control.
In particular, protopanaxadiol-type saponin contents increased by 5.5–9.7 times that of the control, whereas protopanaxatriol-type
saponin contents were increased by 1.85–3.82-fold. In the case of Rg1 ginsenoside after MJ elicitation, the content was affected
negatively in ginseng hairy root cultures.
Plant Cell Tissue and Organ Culture 04/2012; 98(1):25-33. · 3.09 Impact Factor
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ABSTRACT: Cleaved amplified polymorphic sequence (CAPS) marker system using mitochondrial consensus primers was applied for molecular identification of Korean ginseng cultivars (Panax ginseng). Initially, a total of 34 primers were tested to six Korean ginseng cultivars and two foreign Panax species, P. quinquefolius and P. notoginseng. In the polymerase chain reaction (PCR) amplification results, four primers (mt7, mt11, mt13, and mt18) generated co-dominant polymorphic banding patterns discriminating the Korean ginseng cultivars from P. quinquefolius and P. notoginseng. In the CAPS analysis results, the majority of the cleaved PCR products also yielded additional latent polymorphisms between the Korean ginseng cultivars and two foreign Panax species. Specific latent CAPS polymorphisms for cultivar Gopoong and Chunpoong were detected from internal region amplified with mt9 primer by treating HinfI and Tsp509I endonucleases, respectively. Sequencing analysis revealed that the length of amplified region of Korean ginseng cultivars was 2,179 bp, and those of P. quinquefolius and P. notoginseng were 2,178 and 2,185 bp, respectively. Blast search revealed that the amplified region was a mitochondrial cytochrome oxidase subunit 2 (cox2) gene intron II region. Nineteen single nucleotide polymorphisms (SNP) including each specific SNP for Gopoong and Chunpoong, and three insertion and deletion (InDel) polymorphisms were detected by sequence alignment. The CAPS markers developed in this study, which are specific to Gopoong and Chunpoong, and between the Korean ginseng cultivars and two foreign Panax species, will serve as a practical and reliable tool for their identification, purity maintenance, and selection of candidate lines and cultivars.
Molecular Biology Reports 05/2011; 39(1):729-36. · 2.93 Impact Factor
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ABSTRACT: This study describes an efficient approach for developing sequence tagged sites (STS) for Panax ginseng C.A. MEYER, and their applications for line discrimination. By using the methylation filtering (MF) technique, a genomic library was constructed, in which clone inserts were derived from the hypomethylated regions of ginseng genome. A methylation unfiltered genomic library was also constructed and the clone inserts were compared to those from the MF library in terms of sequence characteristics. Sequence analysis revealed that MF efficiently enriched the protein coding region of P. ginseng, for which the repetitive DNA appeared to be as little as 2.5 fold lower than clones in the unfiltered library, and also indicated that the P. ginseng genome may contain a large fraction of methylated repetitive DNA elements. A total of 99 and 100 highly stringent STS primer sets were designed from the filtered and unfiltered library, respectively. Amplification products were tested for latent polymorphism across six cultivars of P. ginseng and other 2 Panax species using six endonucleases recognizing four-bases. STS primer sets described here will be useful for marker-assisted selection, genome mapping and line discrimination of P. ginseng or its cultivars from other Panax species.
Biological & Pharmaceutical Bulletin 01/2010; 33(9):1579-88. · 1.66 Impact Factor
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ABSTRACT: To elucidate the exact function of CabAS in Centella asiatica, which was previously reported as a putative beta-amyrin synthase [Plant Cell Rep, 24:304-311, 2005], this gene was functionally expressed in the lanosterol synthase-deficient yeast mutant (erg7). After inducing the CabAS gene with galactose, a peak consistent with the dammarenediol standard was detected in LC/APCIMS analyses and the accumulated product was confirmed as dammarenediol. CabAS should therefore be renamed to C. asiatica dammarenediol synthase (CaDDS). The confirmation of this gene function may allow us to better understand the generation of numerous triterpene carbon skeletons.
Plant Physiology and Biochemistry 08/2009; 47(11-12):998-1002. · 2.84 Impact Factor
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ABSTRACT: Transformed root ("hairy root") cultures have been shown to be a good model for the study of many secondary metabolites. However, economically important compounds such as asiaticoside and madecassoside are produced in insignificant amounts in the root of Centella asiatica (L.) Urban. To overcome this problem, C. asiatica was transformed using Agrobacterium rhizogenes strain R1000 that harbors pCAMBIA1302 encoding the hygromycin phosphotransferase (hpt) and green fluorescence protein (mgfp5) genes and the hairy culture was coupled with elicitation technique. Hairy roots were obtained at a frequency of up to 14.1% from a tissue junction between the leaf and petiole. Abundant hairy roots were observed when co-cultivation of the plant with A. rhizogenes was done for 7 days (36.1%). Transformation was confirmed by PCR and Southern blot analyses. Five weeks after inoculation, no asiaticoside was detected in the hairy root samples. However, when 0.1 mM methyl jasmonate (MJ) was applied as an elicitor to the culture medium for 3 weeks, a large quantity of asiaticoside was generated (7.12 mg/g, dry wt). In the case of gene expression, 12 h after MJ treatment the expression of the CabAS (C. asiatica putative beta-amyrin synthase) gene in the hairy roots is significantly different from that of the control and this level of transcripts was maintained for 14 days. Our results showed that production of C. asiatica hairy roots could be optimized and the resulting cultures could be elicited with MJ treatment for enhanced production of asiaticoside.
Plant Cell Reports 12/2007; 26(11):1941-9. · 2.27 Impact Factor
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ABSTRACT: Molecular authentication among three Panax species and within cultivars and accessions of P. ginseng was investigated using the DNA sequence in the ribosomal ITS1–5.8S–ITS2 region. Four single-nucleotide polymorphisms were
identified between P. ginseng and other Panax species. In the electrophoresis profile, obtained after digestion with the enzyme TaqI, three fingerprinting patterns were obtained from cultivars and accessions of Panax species. Consequently, this authentication procedure based upon the restriction fragment length polymorphism in the ribosomal
ITS1–5.8S–ITS2 region can now be utilized to differentiate these Panax species as well as major Korean cultivars such as Gopoong and Kumpoong from other cultivars and accessions in Panax species at the DNA level.
Plant Biotechnology Reports 07/2007; 1(3):163-167. · 1.19 Impact Factor
