Morus Folium (Sang-yeop in Korean) is one of the most important Oriental medicinal plants. In Korea, both M. alba and M. cathayana are regarded as the botanical sources for Morus Folium. In order to discriminate M. alba and M. cathayana from their adulterant, M. tricuspidata, mitochondrial NADH dehydrogenase subunit 7 (nad7) intron 2 region was targeted for molecular analysis with universal primers. DNA polymorphisms, including SNP sites, insertions, and deletions, were detected among these three species sequencing data. Based on these DNA polymorphisms, specific primers were designed for the three species respectively. Multiplex PCR was conducted for molecular authentication of M. alba, M. cathayana, and M. tricuspidata with specific primers. The present results indicate that it is possible to identify Morus Folium from its adulterant using mitochondrial nad7 intron 2 region. The established multiplex-PCR system was proved to be effective for identification of Morus Folium. The results indicate that mitochondrial introns can be used for inter-specific polymorphic study, and the described method can be applied for molecular identification of medicinal materials.
The research concerned of the regeneration of plants from embryos obtained from anther cultures of ginseng (Panax ginseng C. A. Meyer). The aim was to determine the influence of the regeneration medium on the efficiency of the regeneration process. We conducted to determine the optimum conditions such as cold pretreatment, plant growth regulators and carbon sources on anther culture of P. ginseng. Highest callus formation rate was obtained when flower buds pretreated at 4℃ for 1 day. Among the treated growth regulators with various degrees of concentration in Murashige and Skoog's (MS) medium, 4.53 μm of 2.4-dichlorophenoxyacetic acid and 4.44 μm of 6-benzylaminopurine gives the most responsive callus with the frequency of 73.89% and 129.53 g of fresh weight. When we used 3-9% of sucrose and maltose among the different kinds and various concentrations of carbohydrates, callus was formed highest 67.29% in the medium with 3% of sucrose. Shoots induced from callus supplemented with 28.9 μm of gibberellic acid and rooted in Gamborg's B5 medium supplemented with 14.7 μm of indole-3-butyric acid.
Ginseng (Panax ginseng) is frequently used in Asian countries as a traditional medicine. The major components of ginseng are ginsenosides. Among these, ginsenoside compound K has been reported to prevent the formation of malignancy and metastasis of cancer by blocking the formation of tumor and suppressing the invasion of cancer cells. In this study, ginsenoside Rb1 was converted into compound K, via secreted β-glucosidase enzyme from the Leuconostoc lactis DC201 isolated, which was extracted from Kimchi. The strain DC201 was suspended and cultured in MRS broth at 37℃. Subsequently, the residue from the cultured broth supernatant was precipitated with EtOH and then dissolved in 20 mM sodium phosphate buffer (pH 6.0) to obtain an enzyme liquid. Meanwhile, the crude enzyme solution was mixed with ginsenoside Rb1 at a ratio of 1:4 (v/v).The reaction was carried out at 30℃ and 190 rpm for 72 hours, and then analyzed by TLC and HPLC. The result showed that ginsenoside Rb1 was transformed into compound K after 72 hours post reaction.
Introduction
The ginseng saponin (ginsenoside) is one of the most important secondary metabolites in ginseng and hasvarious pharmacological activities. To date about 38 kinds of ginsenosides have been isolated and identified from Panax ginseng C. A. Meyer. Among these ginsenosides, Rg3 is a precursor for ginsenoside Rh2, which has a very strong antitumor effect. and has many pharmaceutical activities. However, Rg3 is extremely low in normal ginseng. Thus production of ginsenoside Rg3 would be very important and many studies have aimed to convert major ginsenosides to the more active minor ginsenoside Rg3. The enzymatic conversion through sugar hydrolysis at a specific position is desirable for the production of active minor ginsenoside Rg3.
Material and Method
The isolation of β-glucosidase-producing microorganisms was performed according to a previously published method. Each microbialsuspension cultured in nutrient broth was added to the same volume of 1 mM ginsenoside Rb1 solution and then incubated on a rotary shaker at 30°C for 48 h. The reaction mixture was extracted with butanol saturated with H2O and then analyzed by thin layer chromatography (TLC). 8 μl of the ginseng extract solution was spotted on a TLC plate and developed to 5.5 cm distance in a chamber with chloroform/methanol/water as the mobile phase. Bands on the TLC plates were detected by spraying 10% H2SO4, followed by heating.
Result and Discussion
Ginseng(the root of Panax ginseng C. A. Meyer, Araliaceae) is frequently used as a crude substance taken orally in Korea, China and Japan, as well as other Asian countries, as a traditional medicine. Ginsenosides are the principal components having pharmacological and biological activities. More than 38 different ginsenosides so far have been isolated and identified from ginseng saponins. Among them, deglycosylated ginsenosides are known to be more effective in vivo physiological action and to act as active compounds. A lactic acid bacteria, which have β-glucosidase activity, were isolated from soil and kimchi using a MRS-Esculin agar. These strains were identified on the basis of phylogenetic inference based on 16S rDNA sequences. TLC and HPLC were used to analysis transformed ginsenosides. Ginsenosides are main pharmacoactive component in ginseng. When ginseng was orally administered, the absorption of ginsenosides from the gastrointestinal tract are extremely low. In order to improve oral bioavailability, transforming major ginsenosides into more active minor ginsenoside is very important. Caulobacter leidyia GP45 and Micro- bacterium esteraromaticum GS514 were isolated from ginseng field for converting major ginsenosides into minor ginsenosides. In the co-culture of strain GP45 and GS514 with ginsenoside Rb1, produced compound K and ginsenoside Rg3 individually. The transformation pathway of ginsenoside Rb1 were confirmed Rb1⟶Rd⟶F2⟶compound K and Rb1⟶Rd⟶Rg3.
Ginseng (Panax ginseng C. A. Meyer), a perennial herb in the Araliaceae family, is one of the most commonly utilized medicinal plants in the world. Because whole genome sequence of the ginseng plant is not analyzed, the expressed sequence tags (ESTs) from a ginseng plant was studied by constructing cDNA libraries. Almost 20,000 ESTs were collected and BLAST comparisons of the cDNAs in GenBank’s non-redundant databases revealed that many cDNAs (89.1%) exhibited a high degree of sequence homology to genes from other organisms. The majority of the identified transcripts were found to be genes related with energy, metabolism, subcellular localization, and protein synthesis. Many cDNA clones containing fructose-1,6-bisphosphate aldolase, dehydrin, catalase, and glutathion-S-transferase etc. were analyzed to study their characteristics. The expressions of the genes in different organs were analyzed using reverse transcriptase (RT)-PCR. In addition, the expressions of the genes under different abiotic stresses were analyzed at different time points.
Ginseng Genetic Resource Bank (GGRB) in Kyung Hee University is the center constructed for ginseng genomic and genetic information. GGRB collects genetic resources of ginseng cultivars with systematic management system for effective conservation and application of the resources. GGRB supplies researchers new ginseng cultivars, specific markers, ginseng DNA and RNA for scientific researches.
This study was conducted to determine feasibility of utilization of Angelica acutiloba. Especially, the quality characteristics of bread prepared with the addition of Angelica acutiloba powder were investigated. Sensory evaluation and spoilage test were conducted for preparation of functional breads which added with ground plant matters (leaves and stems) from Angelica acutiloba. The result showed that the functional breads had high score of overall liking as well as low spoilage rate when added with 0.5 to 1.0% ground plant matters of Angelica acutiloba. Consumer acceptability evaluation showed a significant preference when added 0.5 to 1.0% ground leaves and stems of Angelica acutiloba into breads. Functional breads which added powder of Angelica acutiloba inhibited the growth of fungi. The more addition of Angelica acutiloba powder, the higher the degrees of this inhibited. These results suggested that the shelf-lives of the breads were extended by the addition of Angelica acutiloba powder. Further studies were required for improvement of functionality and diversity of bread products using medicinal plant materials as an additive.
A cytosolic ascorbate peroxidase, hydrogen peroxide-scavenging enzyme, was characterized from Codonopsis lanceolata. The cytosolic ascorbate peroxidase cDNA (CAPX) was 983 nucleotides long and possess an open reading frame of 753 bp with 251 amino acids (MW 27.9 kDa) with pI 5.61. The deduced amino acid sequence of CAPX shows high homology to other known cytosolic APXs (78~83%), but the CAPX was clustered independently from compared ten plant APXs. The CAPX gene was highly expressed in leaf and stem tissues, but not in root. When Codonopsis leaves cut using scalpel were soaked in 1 mM hydorgen peroxide, the expression of CAPX gene was suppressed.
A cinnamoyl CoA reductase (CCR) cDNA (ClCCR) was isolated from tabroot mRNAs of Codonopsis lanceolata by cDNA library construction, and its expression was investigated in relation to abiotic stresses. The ClCCR is 1008 bp in length with an open reading frame (ORF) of 336 amino acids. The deduced amino acid sequence was showed high similarity with cinnamoyl-CoA reductases of P. tremuloides (AAF43141) 87%, F.×aranassa (AAP46143) 83%, L. album (CAD29427) 80%, E. gunnii (CAA66063) 72%, S. tuberosum (AAN71761) 83%. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis was revealed that the ClCCR expression was regulated by abiotic stresses.
A thoredoxin (CTRX) gene was cloned and characterized from a full length cDNA library prepared from taproot of three-year old Codonopsis lanceolata. A CTRX was 666 nucleotides long and has an open reading frame of 372 bp with 124 amino acid residues (pI = 4.92). The deduced amino acid sequence of the CTRX matched to the previously reported plant thioredoxin h genes. The deduced amino acid sequence of CTRX exhibited the similarity of 33-67% among previously registered thioredoxin genes. The expression of CTRX in leaves of Codonopsis lanceolata was increased by wounding and 1 mM H2O2, but decreased by 0.1 mM cadmium.
A class I type 2 metallothionein (CMet2) cDNA from taproot of Codonopsis lanceolata was isolated and characterized. A CMet2 cDNA was 572 nucleotides long and had an open reading frame of 234 bp with a deduced amino acid sequence of 78 residues (pI = 4.99). The deduced amino acid sequence of CMet2 matched to the previously reported type 2 metallothionein-like protein genes and showed 74% identity with that of G. max (BAD18377) and C. arietinum (CAA65009). Expression of CMet2 by the RT-PCR was increased at 1 hr after cadmium and hydrogen peroxide treatment, respectively.
This study was conducted to investigate the effect of activated carbon on leaf and stem production of Agastache rugosa as affected by different amounts of activated carbon. The results obtained are summarized as follows. Growth characteristics including plant height and leaf length were the highest when activated carbon added with 10%, suggesting that optimum amount of activated carbon was ranged from 10 to 20%. Growth and enlargement of the root were improved by 10% AC. Activated carbon can be utilized as a soil conditioner in agricultural crop areas.
This study was conducted to determine feasibility of production system of Perilla frutescens leaf-stem by fertilizing of Sta-Green in pots. Germination rate of Perilla frutescens seeds collected in 2002 was 7%, also germination rate of seeds collected in 2003 was 62%, while germination rate of seeds collected in 2004 was above 93%. Seed germination rate of Perilla frutescens collected in 2004 were higher than seed gathering in 2002. Especially, plant growth and yield of Perilla frutescens grown in pot(The pots was filled with soil mixtures of Sta-Green and Peat Moss mixed with 40:60 ratio.) was the highest. These results indicate that leaf and stem production of Perilla frutescens can be improved by fertilizing of Sta-Green in pots.
A full-length cDNA (PPrx1) encoding peroxidase has been isolated and its nucleotide sequence determined from flower bud in ginseng plant (Panax ginseng). A PPrx1 cDNA is 1192 nucleotides long and has an open reading frame of 1062 bp with a deduced amino acid sequence of 354 residues (pI 7.53). The deduced amino acid sequence of PPrx1 matched to the previously reported peroxidase protein genes. The PPrx1 showed a high similarity with the 64% identity with peroxidase of N. tabacum (AAK52084). In the phylogenetic analysis based on the amino acid residues, the PPrx1 was closer with peroxidase of G. max (AAD37376).
Ribosomal protein complex with ribosomal RNA to form the subunits of the ribosome serve essential functions in protein synthesis. A full-length cDNA (PRPS4) encoding ribosomal protein S4 has been isolated and its nucleotide sequence determined in ginseng plant (Panax ginseng). A PRPS4 cDNA is 1105 nucleotides long and has an open reading frame of 792 bp with a deduced amino acid sequence of 264 residues (pI 10.67). The deduced amino acid sequence of PRPS4 matched to the previously reported ribosomal protein S4 genes. Their degree of amino acid identity ranged from 68 to 92%. Phylogenetic analysis based on the amino acid residues showed that the PRPS4 grouped with ribosomal protein S4 of S. tuberosum (CAA54095).
A cDNA, PSOD1, encoding cytosolic copper/zinc superoxide dismutase (CuZn-SOD) was cloned and characterized from a full length cDNA library prepared from Populus alba×Populus glandulosa cultured in vitro. A PSOD1, is 725 nucleotides long and has an open reading frame of 459 bp with 152 amino acid residues (pI 5.43). The deduced amino acid sequence of PSOD1 perfect matched to the previously reported CuZn-SOD (CAC33845.1). Consensus amino acid residues (His-45, -47, -62, -70, -79, -119) were involved in Cu-, Cu/Zn-, and Zn- binding ligands. The deduced amino acid sequence of PSOD1 exhibited the high level of similarity from 100 to 85% among previously registered SOD genes. The expression of PSOD1 in poplar increased at the 1 mM H2O2 and drought stress during 30 min and 60 min, but the ozone treated poplar increased at 30 min in the early time and then decreased at 60 min.
A type 3 metallothionein cDNA (PMT3a) from ozone-treated Populus alba×Populus glandulosa cDNA library has been isolated and characterized. A PMT3a cDNA is 459 nucleotides long and has an open reading frame of 201 bp with a deduced amino acid sequence of 66 residues (pI 4.94). The deduced amino acid sequence of PMT3a matched to the previously reported metallothionein genes. The deduced amino acid sequence of PMT3a showed the 86% identity with P. balsamifera ×P. deltoides. Expression of PMT3a by the RT-PCR was increased 60 min than 30 min after drought treatment. The ozone treated poplar increased at 30 min in the early time and then decreased at 60 min.
Calmodulin, a Ca2+-binding protein, has no enzyme activity. It combines with Ca2+ and makes variable proteins to an active form. Calmodulin 2 is a ubiquitous protein in plants. To investigate the defense mechanism against various stresses, a clone encoding a calmodulin 2 protein was isolated from a cDNA library prepared from taproot mRNAs of Codonopsis lanceolata. The cDNA, designated CICAM2, is 719 nucleotides long and has an open reading frame of 450 bp with a deduced amino acid sequence of 149 residues. The deduced amino acid sequence of CICAM2 showed a high similarity with calmodulins of P. x hybrida (P27163) 97%, N. tabacum (BAB61908) 97%, S. tuberosum (AAA74405) 96%, Z. mays (CAA74307) 92%, C. richardii (AF510075) 93%, M. truncatula (AAM81203) 91%, and G. max (P62163) 91%. The transcriptional expression of the CICAM2 gene, was gradually increased by the CaCl2 treatment. Whereas its expression And it was gradually decreased in the cold stress treatment.ent.
To investigate the defense mechanism against the abiotic stress, a cDNA clone encoding a CuZn superoxide dismutase (CuZnSOD) protein was isolated from a cDNA library prepared from tabroot mRNAs of Codonopsis lanceolata. The eDNA, designated ClSODCc, is 799 nucleotides long and has an open reading frame of 459 bp with a deduced amino acid sequence of 152 residues. The deduced amino acid sequence of ClSODCc matched to the previously reported CuZnSODs. Consensus amino acid residues (His-45, -47, -62, -70, -79, -119 and Asp-82) were involved in Cu-, Cu/Zn-, and Zn- binding ligands. The deduced amino acid sequence of ClSODCc showed high homologies (82%-86%) regardless of species. Expression of ClSODCc by oxidative stress was increased up to 1 h after treatment and declined gradually. Much earlier and stronger expression of ClSODCc was observed in the cold stress treatment.