Background : Codonopsis lanceolata is a perennial plant of Campanulaceae with characteristic flavor and aroma and this plant has saponin, flavonoid, and inulin, which are reported to have physiological activity and antioxidant activity. In contrast, breeding or study of C. lanceolata varieties had not been done for a long time. Genetic polymorphism and phylogenetic relationship analysis of the plants by region of the crops can help the collection of genetic backgroud data for variety development. Methods and Results : In this study, we collected 26 C. lanceolata lines (95 individual plants) from 26 regions in Korea. We genotyped the collected lines using SSR markers developed in the previous study and analyzed the population structure based on the results. Population structures were analyzed using model-based STRUCTURE software (version 2.3.4) using the following parameters: Number of clusters (K) set = 1 to 12; Number of Iterations = 5; Length of Burning Period = 100,000; Number of MCMC (Markov Chain Monte Carlo) Reps after Burnin = 100,000. As a result, Of the 26 collections, were genetically grouped into 6 or 7 groups. Conclusion : The 26 C. lanceolata collections (95 individual plants) were genetically grouped but not grouped by collected regions. These results suggest that C. lanceolata has diverse genetic backgrounds and this data could be used as a basis for genetic polymorphism analysis of Codonopsis species.

Background : The roots of Codonopsis lanceolata have been used as a tonic crude drug and an edible plant in Korea. The plant mainly contains triterpenoid saponins, including codonolaside, codonolasideⅠ-Ⅴ, lancemaside A-G. Their saponins have shown anti-inflammatory effects such as bronchitis and cough, insomnia and hypomnesia. C. lanceolata is well known to affect various pharmacological effects for human health, and its consumption is increasing. Recently, plant and plant-derived products were treated a part of the healthcare system by applying the bioactive phytochemicals. Antioxidant and immune activity substances in food play an important role as a health-protecting factor. This study was designed to investigate the in vitro immune cell growth and xanthine oxidase Inhibitory activity of different storage period and storage temperature of C. lanceolata. Methods and Results : The plant materials were used the roots of C. lanceolata cultivated in Jeju area, Korea. Immune enhancing effect was conducted using T cell and B cell of human immune cells. Each cell incubated for 8 days with the sample extracts compared to the control group, and the immune activation was measured according to the growth of immune cells. The xanthine oxidase Inhibitory activity was measured by modifying the method of Noro(1983). In different storage period and storage temperature conditions, the immune cell growth of C. lanceolata extract promoted a concentration-dependent manner in both human T cell and B cell, and did not show a significant difference. The xanthine oxidase Inhibitory activity of C. lanceolata extract tended to decrease more, depending on the longer the storage period or the higher the storage temperature. Conclusion : These results of this study suggested that the root of C. lanceolata may assist in the potential biological activities, and can be used as a source of human health products.

Background : Codonopsis lanceolata is a flowering perennial climber. The roots are used as medicinal materials or vegetables. C. lanceolata is distributed in India and East Asia such as China, Japan as well as Korea. Recently, demand for C. lanceolata is increasing as a healthy food. In South Korea, this plant is widely cultivated in Gangwon-do province. Although, C. lanceolata is one of the most important medicinal plants in Korea, an elite, inbred line or a variety has not been developed yet. Simple sequence repeat (SSR) marker is a powerful tool for analysis of genetic relationships. In addition, it is a useful tool for studying the non-reference plant genome, due to its even distribution throughout the genome, as well as its high polymorphism between individuals. Methods and Results : We constructed microsatellite-enrichment libraries using C. lanceolata genomic DNA, and obtained a total of 226 non-redundant contig sequences. Routine PCR was performed using gDNA as templates for the polymorphic markers screening. Finally, total 15 polymorphic SSR markers based on C. lanceolata genomic sequences were successfully developed. These markers were applied to 53 C. lanceolata collected from Korea. 103 alleles of the 15 SSR markers ranged from 3 to 19 alleles at each locus, with an average of 6.87. The average of observed heterozygosity and genetic diversity were 0.42 and 0.62, respectively. The average of polymorphism information content (PIC) value was 0.57. The genetic distance value ranged from 0.73 to 0.93, and there was no observed distinct group according to the collecting areas. Conclusion : We developed 15 novel SSR markers from C. lanceolata genomic sequences for further genetic studies. Also, we concluded that the lineage of C. lanceolata collected in Korea has not been established systematically.

The present study was performed to investigate the effects of the colchicine concentrations on chromosome doubling for producing of tetraploid plants of Codonopsis lanceolata, and its effect on plant morphology. A total of 180 individuals germinated from 16 treatment groups, were exposed to various concentrations (0.05-1.0% w/v) of colchicine for different soaking duration (3-24 hour). The highest numbers of tetraploid plants (3) were observed from the lowest concentration of colchicine (0.05%), and one (1) tetraploid plant was obtained from the 0.5% concentration group with a 6 hour treatment. However, no tetraploid individual was observed in any other treatment groups. The plant height of the diploid (18.1 ㎝) was slightly shorter than that of the tetraploid (13.4 ㎝). The fresh weight of the main root in the diploid (0.5 g) was four-fold higher than the tetraploid (2.2 g). The colchicine-treated plant regeneration rate in C. lanceolata was decreased when the plants were subjected to high concentration of colchicine. In particular, the highest number of tetraploid plants (5 and 3) was obtained from the lower concentration (0.05% and 0.1%) of colchicine for 6-hour treatment, which were a higher rate (29.4% and 30%) of regenerated tetraploid plants than other regenerated plants. As in the seed treatment result, the plant height of the diploid was significantly higher (10.4 ㎝) than tetraploid. The higher morphological changes were observed comparatively from tetraploid plants than the diploid.

This study was executed to evaluate the immune activity, nitrite scavenging activity and ABTS radical scavenging activity against extracts of various concentration of ethanol solvent from Codonopsis lanceolata cultured at 6 local regions. The immune responses from both human T and B cell line was significantly enhanced in the cell growth compared to control while the cell growth was influenced at a certain period of culture. The results revealed that the cell growth of both human T and B cell was altered in a time dependent manner. The nitrite scavenging activity of ethanol extracts from various solvent concentration of C. lanceolata were affected by pH. At a pH of 1.2, the nitrite scavenging effect of all of the extracts tested observed higher than that of the other two pH ranges. There was no distinct detection of nitrite scavenging effects of the pH range 6.0. The ABTS radical scavenging activity was progressively increased in a dose-dependent manner, and the activity was the highest in 100% ethanol extract. The result from this investigation suggests that the extract of Codonopsis lanceolata could be an addition to basic medicine for immune modulation and natural food additives.

This study was conducted to investigate the effects of temperature and shade, which are basic environmental conditions, on growth, yield, inorganic components, and general components of Codonopsis lanceolata, in order to obtain basic data for improving yield capacity. In natural light, in the 15, 20, and 25°C groups, the plant heights ranged between 218.9 cm and 223.9 cm, and there was no significant difference between groups. However, the leaf size was larger in shade, and the leaf area was significantly larger in the 15 and 30°C groups. In natural light, root length and diameter were shorter and thinner when the temperature was higher, and growth was highly suppressed at 30°C. With regards to macroelements, the contents of Na, Mg, and P increased as temperature increased, regardless of the plant part; however, no constant tendency was observed in K and Ca according to temperature. The contents of Mg and Ca (from highest to lowest) were in the order leaf>stem>root, whereas the contents of Na, P, and K were in the order stem>leaf>root. Contents of general components varied according to temperature, and were highest at 30°C. While the plant height was increased under the constant 25°C +DIF (Difference between day and night temperature) condition, growth was suppressed in the –DIF group, in which the night temperature was higher than the day temperature, which suggests that a change in night temperature is one of the factors that affects the growth of C. lanceolata. As in the growth of the above-ground parts, fresh weight of the root was high in the constant 25°C group and +DIF group. Notably, it was more than 2.5 times the fresh weights in the constant 15°C group, constant 20°C group, and –15 DIF group.

Background : This study was conducted to determine the effects of mulching, an environmentally friendly organic cultivation method, on antioxidant compounds and growth in Codonopsis lanceolata. Methods and Results : C. lanceolata was treated by mulching with a non-woven fabric, biodegradable film, or rice husks, as well as hand weeding, and non-treatment. The growth and levels of weed control in C. lanceolata were better in plants cultivated under mulching treatments (non-woven fabric, biodegradable film, and rice husks) than under non-mulching treatments (hand weeding, non-treatment). The contents of antioxidant compounds, such as total flavonoids, phenolics, and anthocyanin, were highest under biodegradable film treatment, followed by non-woven fabric treatment, rice husks treatment, hand weeding, and non-treatment. DPPH and ABTS activity were able to identify to have differences their activity in comparison to antioxidant compounds content by solvent fractions. Mulching treatments produced a higher activity in water and ethyl ether fractions for DPPH scavenging activity and in n-butanol fractions for ABTS scavenging activity compared to other fractions, although total activity of DPPH and ABTS were not increased with mulching treatments. Conclusion : Mulching C. lanceolata with biodegradable film and non-woven fabric is an effective method for improving plant growth and inhibiting the occurrence of weeds as well as increasing antioxidant compound content and changing of antioxidant activity.

Background : Codonopsis lanceolata is used as a natural medicine or vegetables. It originates in East Asia such as Korea, Japan and China. C. lanceolata roots contain various chemical compounds including saponins like Panax ginseng. Although C. lanceolata are cultivated in different regions of South Korea, no variety has been developed. Therefore, it is necessary to develop discriminating methods such as molecular markers in C. lanceolata species. Methods and Results : To find simple sequence repeat (SSR) markers, we sequenced C. lanceolata genomic DNA using Illumina HiSeq 2000 System. A total of 250,455 putative SSR loci were obtained, and 26,334 non-redundant primers were designed to amplify these SSRs. Di-nucleotied repeats were the most abundant SSR reapeats, accounting for 89.53% (23,578) of primer designed SSRs. Tri-nucleotide, tetra-nucleotide and penta-nucleotide accounted for 8.44% (2,223), 1.3%, (348) 0.2% (55), respectively. Tri-, tetra-, and penta-nucleotide (total of 2,626 SSRs) were investigated in silico to identify polymorphism between individuals. Consequently, 573 SSRs showed polymorphism. Forty genomic SSR markers were tested in 16 C. lanceolata plants for determination of PCR amplification and polymorphism. From these primers, 27 (67.5%) amplified products and the average polymorphism information content (PIC) value was 0.52. Conclusion : We development 27 SSR markers from C. lanceolata using NGS, and it could be used for breeding of new varieties in the future.

This study was conducted to evaluate the cytotoxicity and α-amylase, α-glucosidase, pancreatic lipase inhibition in vitro by different solvent fractions from the roots of Codonopsis lanceolata. The values of IC50 against Calu-6 cell showed a high effect in n-hexane fraction (10.13 μg/mL) whereas DW fraction exhibited the weakest inhibition on cell viability, having an IC50 value of over 1,000 μg/mL. The values of IC50 against HCT-116 cell showed the highest activity in n-BuOH fraction (102.01 μg/mL), followed by n-hexane fraction (145.85 μg/mL), methylene chloride fraction (332.02 μg/mL), ethyl acetate fraction (462.93 μg/mL) and DW fracion (>1,000 μg/mL). α-Amylase inhibitory activity in methylene chloride fraction and ethyl acetate fraction was found to have a higher inhibitory effect with 24.5% and 25.6% than the other fractions. The highest α-glucosidase inhibitory activity was observed from the ethyl acetate fraction extract, while the extract of DW fraction showed the lowest level of inhibitory activity at given experiment concentration. The pancreatic lipase inhibitory activity of C. lanceolata was found to have a higher the effect in ethyl acetate fraction. Inhibition of lipase activity of the ethyl acetate fraction and n-hexane fraction showed a relatively high, while the extract of DW fraction showed the lowest level at given experiment concentration. These results suggested that the roots of C. lanceolata may assist in the potential biological activity on carbohydrate, lipid Inhibitory activity and anticancer activity.

In this study, genetic diversity of wild Codonopsis lanceolata collected in Korea were analysed using SSR makers. Wild C. lanceolata roots were collected in Jeollanam-do Jangheung-gun Choentae Mountain as in roots. The wild C. lanceolata plants were cultivated in Chungbuk National University greenhouse and the leaves were sampled from 36 plants. The genomic DNA of C. lanceolata was extracted using CTAB. PCR was performed using a program of 35 cycles at 94℃ for 30 sec, 60℃ for 30 sec, and 72℃ for 30 sec with an pre-denaturation of 94℃ for 5 min and a final extension of 72℃ for 30 min. The PCR reaction mixture contains 5 pmole of primers and 20 ng of DNA template in a 20 μL reaction volume. The genotype of the analyzed samples were very different. Therefore, the wild C. lanceolata collected in Korea look genetically diverse.

Codonopsis lanceolata is a perennial climber. The roots are used as medicinal materials or vegetables. Recently, demand for C. lanceolata is increasing as a healthy food. C. lanceolata is distributed in India and East Asia such as China, Japan as well as Korea. In South Korea, this plant is widely cultivated in Gangwon-do province. No C. lanceolata varieties were developed in Korea. The objective of this study is to analyze genetic diversity of C. lanceolata cultivated in Korea using SSR makers. C. lanceolata roots were collected in each region were cultivated in Chungbuk National University greenhouse. Samples were obtained from fresh leaves of 5 plants from each collection region. The genomic DNA was extracted using CTAB. Genetic diversity was analysed using 4 sets of C. lanceolata SSR makers. PCR was performed in total 20 μL reaction volume containing 20 ng of DNA template, 5 pmole of primers. The genotypes of the analyzed samples were very similar. That means that the genetic diversity of C. lanceolata cultivated in Korea is very low.

Codonopsis lanceolata is used as a natural medicine or vegetables. It originates in East Asia such as Korea, Japan and China. Similar to Panax ginseng, C. lanceolata contains saponins as effective components. C. lanceolata is cultivated in many regions of South Korea. But, no variety was developed yet and the origin discrimination in the distribution market of C. lanceolata became a problem. In this study, we collected 20 C. lanceolata regional groups; Hoengseong, Wonju, Samcheok, Chuncheon, Pyeongchang, Hongcheon, Yongin, Yangpyeong, Danyang, Chungju, Bonghwa, Ulleung, Yeongju, Sancheong, Muju, Gwangyang, Sinan, Hwasun, Jeju-si and Seogwipo-si, and tested the genetic relationship using RAPD molecular markers. The genomic DNA was extracted using CTAB and the RAPD analysis was performed using 32 primers of Operon Technologies. NTsys-PC program was used for the phylogenetic analysis of the data.

Asterales are dicotyledonous flowering plants and are one of the Asterid clade, incuding many species as well as Codonopsis and Platycodon. Here, we have determined the complete chloroplast genome sequences of C. lanceolata and P. grandiflorus by using the targeted denovo assembly method of short reads derived from whole genome resequencing. The total lengths of each chloroplast genome sequence are 156,180 bp for C. lanceolata and 155,453 bp for P. grandiflorus. In their chloroplast genomes, 106 genes (75 protein-coding genes, 4 rRNA genes, 23 tRNA genes, and 4 hypothetical chloroplast open reading frames [ycfs]) exhibited the relatively similar positions. Also, 7 protein-coding genes commonly showed to contain introns in both C. lanceolata and P. grandiflorus chloroplast genome, while psaA gene contain intragenic regions only in P. grandiflorus chloroplast genome. In further analysis, we identified the codon usage bias to A or T and found the different simple sequence repeat (SSR) loci of each chloroplast genome (18 SSR loci of C. lanceolata and 16 SSR loci of P. grandiflorus). In the phylogenetic trees based on 72 protein-coding genes, C. lanceolata is more closely related to P. grandiflorus than the other plant species order Asterales. Also, we found the highest sequence diversities of 12 protein-coding genes in small single copy (SSC) region than in the inverted repeat (IRs) and large single copy (LSC) region, and 3 genes such as rpoC2 (LSC region), ndhB (IRs region), and ndhF (SSC region) showed the highest number of segregating sites in each region. Additionally, we developed the molecular markers for phylogenetic applications of C. lanceolata and P. grandiflorus chloroplast genome.

This study was performed to investigate the growth characteristics and inorganic components of Codonopsis lanceolata regarding regional differences. The plant height of Japanese Codonopsis lanceolata was 373.6 cm, so it’s revealed that it has more vigorous growth than Korean won. The flowering time of Korean Codonopsis lanceolata was 2 weeks faster than Japanese one. Total fresh weight of root was 41.0 g and 39.0 g for Korean and Japanese respectively, thus, no significance difference was found. However, regarding fresh weight, Korean one had a more fresh weight (35.4 g) of main root parts, but Japanese one had a more fresh weight (9.6 g) of the lateral root part. Each inorganic component was found more in the aboveground parts, regardless of the region and the content of K was the largest. Regarding the content of macroelements for each part of Codonopsis lanceolata, the content of Na, Mg, P, S, and Ca in Korean Codonopsis lanceolata was found the highest on the leaf, followed by stem and root. In the case of Japanese Codonopsis lanceolata, same result was found on the content of Mg and Ca, however, the highest content of Na and P was found in the stem.

This study was conducted to compare the growth, inorganic components, and proximate components of Codonopsis lanceolata grown in 10 regions of Korea for selecting superior species and breeding by crossing. Among the all tested lines, the shortest plant height (217.12 cm) was observed from the Ulleungdo region line (No. 4) while the longest (273.9 cm) was observed from Hwasun region line (No. 9). In addition, the lines of central and northern region (No. 1~No. 7) tend to have shorter plant height than those of southern region (No. 8~No. 9) except Jejudo region line (No. 10). Flowering tends to be late towards southern region, and lines in central and northern regions were started flowering about 2 weeks earlier than those in southern regions. However, the heaviest root weight was 13.1 g, found in only Jejudo line (No. 10) whereas there was no significant difference found in the other regions which have a range of 8.3~11.0 g. The inorganic components were varied in each line, however, proportion of macroelements, such as K, Ca, and P, was the largest for every line. Especially for Heongseong region line (No. 2), had larger proportion of macroelements than the others. There was a difference of proximate compositions of Codonopsis lanceolata, except the moisture content, among all regions, however, it was generally shown that the content of crude protein (1.31~3.76%) and crude fiber (2.18~3.12%) was the highest.