This study was conducted to elucidate the effect of organic fertilizer on growth characteristics, saponin contents and antioxident activity of Platycodin grandiflorum Ridix roots for organic farming. As basal fertilizers, chemical fertilizer, mixed organic fertilizer, bacterial cultures and fermented oil cake and decomposed manure were treated based on 3 ㎏/10a level before transplanting Platycodin grandiflorum Ridix. In root length, when chemical fertilizer plot and mixed organic fertilizer plot were treated, root length was recorded the highest scores (25.3 and 24.0 ㎝) Root width was recorded the highest score (26.6 ㎝) in chemical fertilizer plot. The number of fine-roots was the highest in mixed organic fertilizer plot and chemical fertilizer plot (20.0 and 17.0), respectively. Fresh weight, which affects directly yield, was the highest in organic fertilizer plot (55.7 g/plant). The content of Platycodin D was shown to 327.4~373.8 ㎎/100 g, the highest values were observed in organic fertilizer plot. The total polyphenol and flavonoid contents were recorded the highest scores (15.5 and 15.3 mg/g, respectively) in organic fertilizer plot. In this study we confirmed that application of organic fertilizer was effective to increase yield and pharmacological effect through increase of the number of fine-roots with high saponin contents.

This study was performed to compare the effect of Platycodi Radix (PG) and Black Platycodi Radix (BPG; developed from fresh Platycodon grandiflorum root by steaming nine times at 80°C for 36 hr and drying nine times at 30°C for 24 hr , at which point it becomes black in color.) on anti-oxidant effect, anti-bacterial properties and ovalbumin (OVA)-induced allergic asthma mouse model by calculating serum cytokines and IgE. To induce the allergic asthma, in the control group and the sample treated group, mice of each group were sensitized intraperitoneally with OVA solution at the 1st, the 7th and the 14th day. Sensitization was performed by aerosol allergen challenges with 1% OVA solution intratracheally at the 21th, the 23th, the 25th and the 27th day. At the 29th day, the mice were killed and the levels of interferon-γ, interleukin-4, 5 and 10, total IgE and OVA-specific IgE were measured. IFN-γ was not different among the PG, BPG, and control groups. IL-4, IL-5, IL-13, total IgE, and OVA-specific IgE were significantly in the BPG group compared with the control group. Considering the above experimental results, this study showed that nine times-repitative steaming process on the roots of Platycodon grandiflorum could enhance reduction of the allergic reaction by reducing level of OVA specific IgE and immune cell infiltration and restoring Th1/Th2 cytokine imbalance.

The present study aimed to compare and investigate the morphological characteristics and yield components according to ploidy level of diploid and tetraploid Platycodon grandiflorum under vinyl-greenhouse and open field conditions. Plant height of diploid and tetraploid P. grandiflorum was 51.3 ㎝, 54.0 ㎝, respectively. The results revealed that the plants grown in the vinyl-greenhouse showed significantly higher growth compared to those grown in the open field. Regardless of the growing place, diploid and tetraploid of P. grandiflorum showed the rapid elongation of internodes after 4 and 3 internodes respectively and elongation tends to be decreased as entering the flower-bud differentiation period. The starting day of flowering in vinyl-greenhouse cultivation was found to be faster than that of the open field cultivation by 2∼ 3 days and tended to be delayed by about 5∼6 days in tetraploid P. grandiflorum compared to diploid. Fresh weight of roots from the vinyl-greenhouse cultivation showed a high quantity as 34.2g and 49.4g in diploid and tetraploid P. grandiflorum, respectively and especially tetraploid P. grandiflorum was found to be increased by approximately 44.4% compared to other plots.

The present study was performed to compare the morpho-physiological characteristics of the tetraploid and diploid varieties of Platycodon grandiflorum and to obtain basic data for cultivating a tetraploid variety with high yield and content of functional substances. The plant height of the tetraploid variety (54.0 cm) was slightly higher than that of the diploid variety. The leaf length and width of the tetraploid variety were 10.2 cm and 7.3 cm, respectively. The results obtained from the present study revealed that the form of the leaf changed from lanceolate to ovate, and the chlorophyll content in the tetraploid variety (16.7) was slightly higher than that in the diploid variety. The photosynthetic rate significantly increased (24%) to 13.4 μmol CO2·m -2 ·s -1 in the tetraploid variety from that of the diploid variety. The pollen viability of the tetraploid variety was decreased by approximately 33% with respect to that of the diploid variety, but this did not have a significant adverse effect on seed production. The fresh weight of tetraploid P. grandiflorum was 49.4 g, which was approximately 44% higher than that of the diploid variety.

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 : Platycodon grandiflorum root(PGR) was one of the primary herbs used in a phlegm-relieving herb from the past. Purified platycoside compounds from the roots of PGR may exhibit neuroprotective, antimicrobial, anti-inflammatory, anti-cancer, anti-allergy, improved insulin resistance, and cholesterol-lowering properties. To evaluate preference and functionality of PGR extracts, PGR was fermented by several lactic acid bacteria. Lactic acid bacteria used were Leuc. mesenteroides N12-4 and N58-5, L. plantarum N76-10 and 56-12, L. brevis N70-9 and E3-8. Methods and Results : This study was performed in order to investigate the changes of platycosides, as well as the antimicrobial activities on bronchus diseases inducing bacteria(C. diphtheriae, K. pnneumoniae, S. aureus, S. pyogenes) of Platycodon grandiflorum root(PGR) fermented by using lactic acid bacteria(Leuc. mesenteroides N12-4, Leuc. mesenteroides N58-5, L. plantarum N76-10, L. plantarum N56-12, L. brevis N70-9, L. brevis E3-8). Growth of L. plantarum on PGR was the most active during lactic acid fermentation by some different strains. Total platycoside, platycoside E, platycodin A, polygalacin D2, polygalacin D and diapioplatycoside E contents of PGR fermented for 96 hours at 37℃ by Leuc. mesenteroides and L. plantarum were increased, while platycodin D and platycodin D3 were decreased. The antimicribial activity on PGR fermented by L. plantarum N56-12 exhibited a strong microbial proliferation in all four kinds of bronchus diseases inducing bacteria and was higher than non-fermented PGR extract. Conclusion : Thus, this results showed antimicrobial activities on bronchus diseases inducing bacteria and platycosides content of PGR by L. plantarum N56-12 were higher than non-fermented PGR extract.

Plant breeding requires the collection of genetically diverse genetic resources. Studies on the characteristics of Platycodon grandiflorum resources have not been carried out so far. The present study was carried out to discriminate P. grandiflorum based on morphological characteristics and genetic diversity using simple sequence repeat (SSR) markers. Methods and Results :We collected 11 P. grandiflorum cultivars: Maries II, Hakone double white, Hakone double blue, Fuji white, Fuji pink, Fuji blue, Astra white, Astra pink, Astra blue, Astra semi-double blue and Jangbaek. Analyses of the morphological characteristics of the collection were conducted for aerial parts (flower, stem and leaf) and underground parts (root). Next, the genetic diversity of all P. grandiflorum resources was analyzed using SSR markers employing the DNA fragment analysis method. We determined that the 11 P. grandiflorum cultivars analyzed could be classified by plant length, leaf number and root characteristic. Based on the genetic diversity analysis, these cultivars were classified into four distinct groups. Conclusions : These findings could be used for further research on cultivar development using molecular breeding techniques and for conservation of the genetic diversity of P. grandiflorum. Moreover, the markers could be used for genetic mapping of the plant and marker-assisted selection for crop breeding.

This study was designed to improve the sensory characteristics and mitigate the bitter taste of Platycodon gradiflorum. It was aimed at investigating the qualitative properties of fermented P. gradiflorum after repeated steaming and drying treatments. P. gradiflorum was heated for 2 hr at 95℃, being the first and third treatments compared afterwards. Lactobacillus plantarum, at a concentration of 10%, was used as starter culture. As a result, the third steaming process and the addition of starter improved the physical and chemical properties of P. gradiflorum i.e., crude saponin and total polyphenol contents increased significantly. Moreover, P. gradiflorum steamed three times and fermented by L. plantarum showed the higher overall preference score. Our results indicated that the three-time steaming and drying was an effective manufacturing process for the production of high-quality fermented P. gradiflorum. Lactic acid-fermented P. gradiflorum also could have a potential use as a valuable resource for the development of functional products.

Balloon flower (Platycodon grandiflorum A. DC.) is a perennial plant of mainly Campanulaceae family, which have been widely used as a food ingredient and herbal medicine in East Asia. Although demands on related products and yearly cultivation area for balloon flower are increasing, diverse fundamental technologies and molecular breeding studies are not very well supported in Platycodons. In this study, 30 random amplification of polymorphic DNA (RAPD) primers were test in an attempt to explore genetic diversities. In addition, sequences information of the actin gene, a well conserved gene encoding a globular protein that forms microfilaments, was retrieved and analyzed. Two actin homologs were recovered; 3.4 kb fragment is a Pg-actin and 1.4 kb fragment is a Pg-actin homolog with 28.6% similarity. We have confirmed that the Pg-actin gene is configured into 4 exons and 3 introns. A single nucleotide polymorphism (SNP), G↔A, was detected on the intron 3, which served as a target for the CAPS marker development. The marker Pg-Actin-Int3 was applied to 32 balloon flower accessions. Balloon flower DNA sequence information generated in this study is expected to contribute to the analysis and molecular breeding and genetic diversity analysis of balloon flowers.

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.

The roots of Platycodon grandiflorum species either dried or fresh, are used as an ingredient in salads and traditional cuisine in Korea. To interpret the root proteins, a systematical and targeting analysis were carried out from diploid and tetraploid roots. Two dimensional gels stained with CBB, a total of 39 differential expressed proteins were identified from the diploid root under in vivo condition using image analysis by Progenesis Same Spot software. Out of total differential expressed spots, 39 differential expressed protein spots (≥ 1.5-fold) were analyzed using LTQ-FTICR mass spectrometry. Except two proteins, the rest of the identified proteins were confirmed as down-regulated such as Isocitrate dehydrogenase, Proteasome subunit alpha type-2-B. However, the most of the identified proteins from the explants were mainly associated with the oxidoreductase activity, nucleic acid binding, transferase activity and catalytic activity. The exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.

Platycodon grandiflorum A. is a perennial plant belongs to Campanulaceae family. This plant has been used herbal medicine ingredient in East Asia. Because of the high saponin content, it is an economically important medicinal plant in Korea. It has been reported that saponins of P. grandiflorum were mainly synthesized in root tissues. The studies about root growth of the plant were few. Expansin is an important protein playing a role in root growth of plants, and is known as a nonenzymatic protein. Expansins are novel plant cell wall loosening proteins leading to turgor-driven cell extension. Expansin encoding genes exist in multigene family, and there are more than 30 genes in Arabidopsis thaliana. and more than 50 genes in Oryaza sativa. Therefore, identification of the genes was difficult in P. grandiflorum because of the lack of genome sequence. Recently, the development of next generation sequencing (NGS) technologies make it possible to obtain the target genes sequences rapidly and precisely. In this study, to identify the expansin encoding genes in P. grandiflorum, we used RNA-seq analysis with Illumina HiSeq platform. We analyzed whole transcriptome of P. grandiflorum through the RNA-seq analysis based on next generation seuqencing. CLC Genomics Workbench software (Clc Bio inc.) was used for assembly. We assembled 122,663 contigs and search 123 contigs were identified from the search using 61 expansin gene

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 perennial herbal plant belongs to Campanulaceae family. It has very important genetic value as a major plant in Asterids order. The major ingredients are platycosides, terpenoid saponins. In Korean industrial plants market, it was produced 5,633 tons in 2013, and the total amount of production was less than only five species, omija, ginger, raspberry, yam and deodeok. P. grandiflorum is called ‘Gilgyung’ and is used as a fresh vegetable and an ornamental plant. Nowadays, various components of P. grandiflorum were already published. But, genetic research is in the starting stage. In this study, 11 cultivars; 1. MariesⅡ, 2. Hakone double white, 3. Hakone double blue, 4. Fuji white, 5. Fuji pink, 6. Fuji blue, 7. Astra white, 8. Astra pink, 9. Astra blue, 10. Astrasemi double blue, 11. Jangback, were analyzed using 60 Operon Universal RAPD primers. The results were phylogenetically analyzed and related to the morphological characteristics of the cultivars.

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.

The roots of Platycodon grandiflorum are massively used in traditional herbal medicine as a remedy for pulmonary disease and respiratory disorders. However, in spite of its potential medicinal significance, the molecular mechanism of its roots is still unknown. In the present study, high throughput proteome approach was conducted to profile proteins from 3, 4 and 5 months aged diploid and tetraploid roots of Platycodon grandiflorum. Two dimensional gels stained with CBB, a total of 68 differential expressed proteins were identified from diploid root out of 767 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 29 differential expressed protein spots (≥ 2-fold) were analyzed using LTQ-FTICR MS whereas a total of 24 protein spots were up regulated and 5 protein spots were down-regulated. On the contrary, in the case of tetraploid root, a total of 86 differential expressed proteins were identified from tetraploid root out of 1033 protein spots of which a total of 39 differential expressed protein spots (≥ 2-fold) were analyzed using LTQ-FTICR MS whereas a total of 21 protein spots were up regulated and a total of 18 protein spots were down-regulated. It was revealed that the identified proteins from the explants were mainly associated with the nucleotide binding, oxidoreductase activity, transferase activity. In that way, the exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.

The roots of Platycodon grandiflorum are known as traditional medicine, has been extensively used since ancient times as a therapeutic to treat cold, cough and asthma in Korean traditional medications. This study was conducted in order to profile proteins from the hormone induced diploid and tetraploid roots using high throughput proteome approach. Two dimensional gels stained with CBB, a total of 64 differential expressed proteins were identified from the diploid root using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 20 differential expressed protein spots (≥ 2-fold) were analyzed using MALDI-TOF-TOF mass spectrometry whereas a total of 13 protein spots were up regulated and 7 protein spots were down-regulated. However, in the case of tetraploid root, a total of 78 differential expressed proteins were identified from tetraploid root of which a total of 28 differential expressed protein spots (≥ 2-fold) were analyzed by mass spectrometry whereas a total of 16 protein spots were up regulated and a total of 12 protein spots were down-regulated. However, proteins identified using iProClass databases revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding, oxidoreductase activity, transporter activity and isomers activity. The exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.

This experiment was conducted to obtain the have higher contents of pharmaceutical constituents as well as higher yield from colchicine induced diploid and tetraploid extracts of Platycodon grandiflorum. In order to determine the biological activity, this study was focused to evaluate the cytotoxicity, antimicrobial on the bronthus disease bacteria, antioxidant enzyme activity of diploid and tetraploid extracts in P. grandiflorum. The activities of antioxidant enzyme according to different solvent extracts were measured as superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX). The cytotoxicity of methanol extracts of P. grandiflorum showed significant differences between tetraploid and diploid. That is, the cytotoxic effect against human cancer cell was higher in tetraploid than in diploid. At all extracts concentration, tetraploid samples showed high toxicity and the IC50 (concentration causing 50% cell death) value showed the highest on HCT-116 cell (105.91 μg/mL), and exhibited significant activity against the Hep 3B cell (140.67 μg/mL), SNU-1066 cell (154.01 μg/mL), Hela cell (158.37 μg/mL), SNU-601 cell (182.67 μg/mL), Calu-6 cell (190.42 μg/mL), MCF-7 cell (510.19 μg/mL). Antimicrobial activities of diploid P. grandiflorum were relatively low compared to tetraploid P. grandiflorum on most of the bacterial strains. In tetraploid P. grandiflorum, K. pneumoniae showed the clear zone formation (18~19 mm) of growth inhibition, followed by the clear zone formation of 13~15 mm on C. diphtheria and S. pyogenes. The antimicrobial activities in diploid P. grandiflorum were the highest on K. pneumonia (14~15 mm), and showed the clear zone formation of 11~12 mm on C. diphtheria and 12~13 mm on S. pyogenes. The antimicrobial activity is thought to look different depending on the bacterial strains and the polyploidy of P. grandiflorum. The root extract of P. grandiflorum had the highest (97.2%) SOD enzyme activity in ethyl acetate partition layer of tetraploid while water partition layer of diploid showed the lowest (48.6%) SOD enzyme activity. The activity of CAT showed higher values in the root of tetraploid than in the diploid of P. grandiflorum in all partition layers except butyl alcohol. The activities of APX and POD showed higher values in the root of tetraploid than in the diploid of P. grandiflorum in all fraction solvents except water layer. These results indicate that the tetraploid P. grandiflorum can be used as a source for developing cytotoxic agent and antimicrobials which can act against bronchus diseases bacterial strains.