Background : Even though Kalopanax septemlobus has been used as a traditional crude drug and a dietary health supplement, the wild or cultivated sources of this plant are not economically feasible. Methods and Results : In this study, a cell suspension culture of K. septemlobus using friable calli was established to make source sustainable. A cell suspension culture of K. septemlobus was incubated during 15th day and reached the maximum capacity of saponin production after day 6 (0.42㎍/㎎ of fresh weight). To investigate the effect of elicitors on the product yield of saponins in the K. septemlobus suspension culture, we treated methyl-jasmonic acid and coronatine (COR), which are known as signal molecules. COR positively regulates the total saponin production in the cell suspension of K. septemlobus at a concentration of 1 μM compared with the mock-treated control. Furthermore, the expression of beta-amyrin synthase (KsbAS) was induced by elicitation of COR. Consequently, oleanane-type triterpene saponins, oleanolic acid (2.369 ± 0.98 ㎍/㎎ of extract) were accumulated by only COR. Conclusion : From the above results, COR is an efficient elicitor for inducing saponin biosynthesis in a K. septemlobus suspension culture. Gene expression pattern and analysing precursor of triterpenoid saponin indicate that major target gene of COR in K. septemlobus suspension culture is KsbAS.

Background : Codonopsis lanceolata is currently used as vegetable, as well as materials for traditional medicines. However, consumers have nagative views on using pesticides and chemical fertilizer in C. lanceolata cultivation. Therefore, this research was conducted to select the appropriate organic fertilizer to improve the growth and saponin components of C. lanceolata by some organic fertilizers application. Methods and Results : Organic fertilizers were applied as 4 types: mixed organic matter, fermentation cake, bacterial culture and rice husks, excluding conventional chemical fertilizer and non-treatment used as control. The result analyzed in soil after fertilizer application showed that soil pH was acidified in fermentation cake and chemical fertilizer treatment, especially, chemical fertilizer treatment showed very high phosphoric-acid content than other treatments, and total N content was higher in fermentation cake, mixed organic matter and chemical fertilizer. Growth of C. lanceolata showed superior tendency in the treatment of mixed organic matter and fermentation cake. Lancemacides could be identified as foetidissimoside A, lancemaside A, lancemaside B, and lancemaside D. However, among them, quantitative analysis could not be conducted on foetidissimoside A due to its very low content, and lancemaside A was the most abundant saponin in the root from all the treatments. The content of lancemaisde A according to organic fertilizer application showed the highest value in the treatment of mixed organic matter, followed by the fermentation cake, bacterial culture, non-treatment, rice husks and chemical fertilizer, in that order. The content of lancemaside B and D was very low compared to lancemaside A, and there was no difference among treatments. Conclusion : The growth of C. lanceolata was superior in application of mixed organic matter and fermentation cake, and the major saponin, lancemaside A, was also increased.

Background : Platycodon grandiflorum is a perennial plant and a member of Campanulaceae family. Since ancient times, they have been using P. grandiflorum as an important medicinal plant in Korea. Platycodin D is the most abundant saponin derived from P. grandiflorum and pharmacologically active component. Cytochrome P450s (CYPs) are important enzymes in the saponin biosynthesis. CYP is, in general, the terminal oxidase enzymes and essential roles in saponin biosynthesis pathway by hydroxylation or oxidaition of triterpene skeletons. Methods and Results : We tried to identify CYP genes related to saponin biosynthesis of P. grandiflorum through RNA-seq analysis. The sequencing was performed using Illumina Hi-Seq platform after cDNA library preparation. The produced reads were assembled using CLC Genomics Workbench software (CLC Bio, Inc.). We obtained 122,663 contigs and found 191 putative CYP genes. Familes of CYP716, CYP708, CYP93 and CYP51 were selected as putative saponin biosynthesis related gene families using phylogenetic relationship analysis. Conclusion : The results in this study could help to find the CYPs related to saponin biosynthesis pathway of P. grandiflorum.

Background : For increasing saponin content of ginseng cultivated in shaded plastic house, this research was performed to investigate growth characteristics and saponin content of ginseng according to foliar spray of chitosan and water-soluble silicates processing. Methods and Results : The ginseng cultivars to be used this research were Cheonpung, Yeonpung and 1-year-old seedlings of Cheonpung, Yeonpung was transplanted on March 24, 2015. Planting density was 72 plant/3.3㎡and Shading material of plastic house used blue-white shading vinyl. The processing method of organic matter is as follows, we were applied chitosan (40 kg / 10a), silicate (3kg / 10a) to soil scheduled for ginseng cultivation with basal application. Then, we were diluted with chitosan and silicates to 1000-fold and investigated growth characteristics, saponin content of ginseng after foliar spray twice a month from May to September. Conclusion : Photosynthetic rate of ginseng according to treatment of organic matter was higher in the chitosan experimental group compared to the control group and photosynthetic rate of ginseng cultivars was the highest in Cheonpung’s silicate group(3.70 μmole CO2/㎡/s). The growth characteristics in above-ground part of 2-year-old ginseng was a good in the organic matter treatments compared to the control group. Above-ground part’s growth characteristics of ginseng according to treatment of organic matter showed a good trend in plant height, stem length, stem diameter of chitosan experimental group. The growth characteristics in under-ground part of 2-year-old ginseng was a good in the organic experimental groups. In Yeonpung of the chitosan experimental group, fresh root weight per plant was 9.2 g/plant that increased by 46 % compared to the control group. In Cheonpung of the silicate experimental group, fresh root weight per plant was 8.3 g/plant that increased by 56 % compared to the control group. In the chitosan experimental group, crude saponin content of under-ground part was Cheonpung(10.38 ㎎/g), Yeonpung(12.29 ㎎/g), control(11.53 ㎎/g), respectively. In the silicate experimental group, crude saponin content of under-ground part was Cheonpung(11.73 ㎎/g), Yeonpung(14.21 ㎎/g), respectively. Crude saponin content of ginseng according to treatment of organic matter was. generally higher in Yeonpung than Cheonpung.

Background : Angelica gigas, also called Dang Gui or Korean Angelica, is a major medicinal herb used in Asian countries such as Korea, Japan and China. A. gigas has many active constituents such as dercursin, decursinol angelate, nodakenetin, nodakenin, β-sisterol or α -pinene. But, there is no research on the gexpression of the genes related to saponin biosynthesis from A. gigas. In this study, we compared the expression of saponin biosynthesis related genes from various organs of A. gigas. Methods and Results : The reads of Angelica gigas mRNAs were produced using Illumina Hiseq 2000, and the reads were assembled to produce 113,597 contigs using CLC　Genomic Workbench. To select the saponin biosynthesis genes, assembled contigs were subjected to BLAST analysis at NCBI site. RNAs were extracted from five tissues, roots, stems, flowers, old leaves and young leaves of A. gigas. We produced total of 16 gene specific primers and used for RT-PCR. PCR conditions composed pre-denaturation at 95℃ for 3min, then 35 cycles of 95℃ for 30 sec, 57℃ for 30sec and 72℃ for 1min, and a final extension at 72℃ for 5min. Electrophoresis performed at 100 V, 30 min using 1.2% gel. Our experiment shows that A. gigas has several genes related to saponin biosynthesis and the genes were expressed from variety of organs. Conclusion : From the above results, we may suggest A. gigas genes related to the biosynthesis of saponins.

Background : Platycodon grandiflorum is a perennial plant and a member of Camanulaceae family. Since ancient times, they have been using P. grandiflorum as an important medicinal plant in Korea. Platycodin D is the most abundant saponin derived from P. grandiflorum and pharmacologically active component. UDP-glycosyltransferases (UGTs) are important enzymes in the saponin biosynthesis. UGT is a glycosyltransferase and act on the final step of the secondary metabolite biosynthesis. Methods and Results : We tried to identify UGT genes related to saponin biosynthesis of P. grandiflorum through RNA-seq analysis. The sequencing was performed using Illumina Hi-Seq platform after cDNA library preparation. The produced reads were assembled using CLC Genomics Workbench software (CLC Bio, Inc.). We obtained 122,663 contigs and found 137 putative UGT genes. Familes of UGT71, UGT73, and UGT74 were selected as putative saponin biosynthesis related gene families using phylogenetic relationship analysis. qPCR condition about UGT73 is preheating 94℃ 180 sec, denaturation 94℃ 60 sec, annealing 53℃ 60 sec, extension 72℃ 90 sec, final extension 72℃ 600 sec, 45 cycles repeated. Conclusion : The results in this study could help to find the UGTs related to saponin biosynthesis pathway of P. grandiflorum.

Background : For increasing saponin content of ginseng cultivated in shaded plastic house, this study was performed to investigate growth characteristics and saponin content of ginseng (Panax ginseng C.A. Mayer) according to foliar spray of germanium and water-soluble silicates processing. Methods and Results : Used a native species in this study is violet-stem variant that most commonly cultivated in ginseng’s farms. 1-year-old violet-stem variant was transplanted on March 24, 2015 and planting distance was 11 × 20 cm. Shading material of plastic house was used blue-white shading vinyl. The processing method of inorganic dissolved matter is as follows, we were diluted with germanium and water-soluble silicates to 500-fold, 1000-fold and investigated after foliar spray twice a month from May to September. The growth characteristics in above-ground part of 2-year-old ginseng was a good in the inorganic matters treatment compared to the control treatment, the difference of above-ground growth characteristics between the inorganic dissolved matters treatments was not significant. The growth characteristics in under-ground part of 2-year-old ginseng was a good in the inorganic matter treatments and root weight per plant in the 500-fold dilution of germanium was 12.4 g that increased by 29 % compared to the control (9.6 g). Crude saponin content of under-ground part was higher generally in inorganic matter treatments compared to the control (11.53 ㎎/g). In the 1000-fold dilution of water-soluble silicate and germanium (50:50), crude saponin content was the highest in 12.91 ㎎/g. Crude saponin content of above-ground part was higher generally in inorganic matter treatments compared to the control (61.76 ㎎/g). In the 1000-fold dilution of water-soluble silicate and germanium (50:50), crude saponin content was the highest in 65.69 ㎎/g. Conclusion : From the above results, we concluded that germanium and water-soluble silicates could be useful matters in promoting growth characteristics and saponin content of ginseng (Panax ginseng C.A. Mayer).

Background : Korean Red Ginseng has been reported to have various anti-virus, anti-tumor, anti-inflammation and immune-regulating activity. However, Korean Red Ginseng Crude Saponin (KRGCS) effect on Pulmonary inflammation has been insufficient research. We have investigated the effect of KRGCS on pulmonary inflammation in mouse model. Methods and Results : Pulmonary inflammation induced by cigarette smoke condensates (CSC) and lipopolysaccharide (LPS) in BALB/c mice. 100uL (ratio 1:1) of CSC (4mg/mL) and LPS (100ug/mL) solution instilled with intratracheal injection once per a week for 3 weeks in mice. KRGCS was also administered orally a dose of 10mg/kg and 25mg/kg for 3 weeks. KRGCS treatment groups decreased the number of inflammatory total cells in lung tissues and bronchoalveolar lavage fluid (BALF) and the number of neutrophils in BALF dropped more than negative control groups. Additionally, KRGCS reduced the absolute number of CD4+ and CD8+ cells in BALF, and the number of CD11b+ Gr-1+cells in the lung and BALF. Furthermore, we found that KRGCS declined significantly the level of the cytokine chemokine (C-X-C motif)-1 (CXCL-1), Macrophage inflammatory protein-2 (MIP-2), and tumor necrosis factor alpha (TNF-α) in BALF. Conclusion : These results that KRGCS has an inhibitory effects on pulmonary inflammation such as COPD, may be a candidate of effective herbal pharmaceuticals for the cure of lung diseases.

Background : Platycodon grandiflorum radix used as a therapeutic agent for lung and respiratory diseases in traditional Korean medicine. The saponin and sugar content in P. grandiflorum root is dependent on the drying temperature and method. In this study, we investigated saponins and antioxidant contents in dried P. grandiflorum roots either not cut or cut to different lengths. Methods and Results : P. grandiflorum roots were either cut to 0.5, 1.0 or 2.0 ㎝ in length or left uncut. They were dried at 45℃ and the effect of antioxidant substances, antioxidant ability, and saponin content were investigated. P. grandiflorum roots rapidly dried out when cut to short lengths. The saponin contents (platycodin D, polygalacin D, and deapioplatycodin D) was highest in the uncut P. grandiflorum roots. When the cut lengths were long, the levels of polyphenolic compound increased. However, 2,2'-azinobis- 3-ethylbenzo-thiazoline-6-sulfonic acid (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities were higher in the samples cut to shorter lengths. Conclusions : These results suggested that the saponin and antioxidant content of P. grandiflorum roots could be maximized by controlling the drying rate of the roots via adjustments to the root cutting length.

Soybean [Glycine max (L.) Merr.] seeds are abundant in high-quality proteins and fats. In addition, soybean seeds are also rich in secondary metabolites, such as isoflavones, lecithin, and saponins. Triterpene saponins are major components of these physiologically active metabolites in soybean seeds. Soybean saponins are classified as group A and DDMP saponins. Among them group A saponins are undesirable component of food products due to bitterness and astringency and also cause foaming in tofu production. Whereas, DDMP saponins and their derivatives are less bitter and astringent and beneficial to human health when consumed as regular diet. Therefore, reducing the group A saponins or increasing the DDMP saponins are required to improve the food quality. The present study focused to identify and characterize the gene which is encoding a protein responsible for biosynthesis of DDMP saponins. EMS mutant lines (sg-7-1 & sg-7-2) which lack DDMP saponins were developed. The breeding cross has been made with these two mutants with two cultivars, Pungsannamul and Wooram to study the segregation and genetic linkage analysis, respectively. The segregation analysis showed that the mutant phenotype is controlled by single recessive gene. TLC analysis for phenotyping F2 population of Wooram X sg-7-1 showed mutant, wild and heterozygous types. To surprise two more patterns were detected and they were named as strange type1 (ST1) and strange type2 (ST2). Further, SSR marker analysis will be carried out to locate the gene which encoding a protein responsible for biosynthesis of DDMP saponins.

Platycodon grandiflorum is a herbal flowering perennial plant belongs to Campanulaceae family. The saponins derived from P. grandiflorum were termed platycosides and platycodin D, which is the most abundant saponin in the plant and pharmacologically active component, was intensively studied. Platycodin D is synthesized from triterpenoids by several enzymes including cytochrome P450. Cytochrome P450 is known to exist in superfamily in plant kingdom and essential roles in saponin biosynthetic pathway by hydroxylation or oxidation of triterpene skeletons. However, the key genes of P450 involved in biosynthesis of saponin was not identified because of its low conservation rate in amino acid sequence level among plant species and gene superfamilies. Recently, next generation sequencing (NGS) technology is rapidly developed as a method to discover target genes. In this study, we tried to identify P450 genes involved in saponin biosynthetic pathway from the various tissues of P. grandiflorum using RNA-seq analysis. The sequencing was performed using Illumina Hi-Seq platform after cDNA library preparation. The produced reads were assembled using CLC Genomics Workbench software (CLC Bio, Inc.). We obteined 122,663 contigs and found out 191 putative P450 genes. The phylogenetic relationship was analyzed and putative genes related to platicoside biosysthesis were selected and cloned for further analysis.

Platycodon grandiflorum is a species of herbaceous flowering perennial plant of the family Campanulaceae. The major ingredients are platycosides, terpenoid saponins. It contains 1-4 % of the dry weight and there are about 20 types of platycosides. Among them, platycodin D have various pharmacological effects on cough and cold. Platycosides are synthesized from oleanane by mevalonic acid pathway and cytochrome P450s and UGTs are important enzymes in the saponin biosynthesis. UGT is glucose transfer enzyme and act on the final step of the secondary metabolite biosynthesis. In this study, we tried to identify UGT genes involved in saponin biosynthetic pathway from the various tissues of P. grandiflorum and non germinated seeds using RNA-seq analysis. The sequencing was performed using Illumina Hi-Seq platform after cDNA library preparation. The produced reads were assembled using CLC Genomics Workbench software (CLC Bio, Inc.). We obtained 122,663 contigs and found 137 putative UGT genes. The phylogenetic relationship was analyzed and putative genes related to platicoside biosysthesis were selected and cloned for further analysis.