This study was to investigate the improvement of immune activities of the extracts from Codonopsis lan-ceolata by stepwise steaming process and high pressure process. The phenol contents was 8.742㎍/㎎ which was higher thanthat from conventional extraction using 70% ethyl alcohol at 80℃ for 24hours. All of extracts at a concentration of 1.0㎎/㎖showed relatively low cytotoxicity on human normal kidney cell (HEK293) in range of 16 19%. The immune B and T cellgrowth was improved by extracts using the steamed and high pressure precess of C. lanceolata up to 180×10⁴cells/㎖ and96×10⁴cells/㎖, respectively. The extract prepared also greatly increased the secretion of both IL-6 and TNF-α from thestepwise steamed and high pressure process. This results can conclude that stepwise steamed and high pressure processeffectively released active biomaterials which could important role in enhancing immune activity in the body.

Codonopsis lanceolata (Campanulaceae) has been used in traditional medicines, as its roots contain severalkinds of 3,28-bidesmosidic triterpenoid saponin with high medicinal values. In this study, we induced hairy root-derivedtransgenic plants of C. lanceolata and analyzed triterpenoid saponins from the leaf, stem and root. Transgenic plants wereregenerated from the hairy roots via somatic embryogenesis. The saponins are lancemaside A, B and E, foetidissimoside A,and aster saponin Hb. Transgenic plants contained richer triterpenoids saponin than wild-type plants. Major saponin lance-maside A was the most abundant saponin in the stem from transgenic-plant, 4.76㎎·1−¹ dry stem. These results suggest thattransgenic plants of C. lanceolata could be used as medicinal materials for the production of triterpene saponins.

세척방법 차이에 따른 신선편이 슬라이스 더덕의 저장 중 품질 변화를 알아보기 위하여 10℃에서 저장 실험을 진행하였다. 저장 10일 후에 무세척구와 손세척은 전체의 80.3% 더덕이 부패되어 상품성을 상실하였을 뿐만 아니라 조직감의 연화도 급격히 진행되어 측정이 불가능하였다. 그러나 버블세척구들은 20.4∼30.1%의 중량 손실만을 보 였으며, 특히 마이크로버블세척구는 저장초기에 비하여 11.1%정도의 조직감만 감소하여 저장 중 변화가 가장 적었 다. 슬라이스 더덕의 표면색도 갈변화를 최소화시키며 저 장 중 색을 유지하는데 효과적인 것으로 나타났다. 저장 중 수분함량의 변화는 모든 처리구에서 감소하였는데, 특 히 무세척구와 손세척은 각각 74.6, 78.3%로 수분함량이 크게 감소하였다. 저장 중 미생물의 수도 마이크로버블세 척, 일반버블세척, 손세척, 무세척구 순으로 증식하는 것으 로 나타났다. 관능검사와 video microscope system을 이용하 여서도 마이크로버블 세척구는 이물질이 발견되지 않았음 을 알 수 있었으며, 다른 처리구들보다 세척효과도 우수한 것으로 확인되었다. 이는 마이크로버블세척을 하여 슬라이스 더덕을 저장하는 것이 품질유지에 가장 적합하였 으며, 저장수명도 10일(10℃ 저장시)로 가장 길게 나타났다.

In general, stepwise hot steaming process is known to be effective in improving its biological activities; however, not much employed in processing Codonopsis lanceolata due to its hardness. The complex processed C. lanceolata showed highest free radical scavenging acitivity as 45.21%. Total phenol and flavonoid content were of complex processed C. lanceolata higher than conventional extract and alone steaming process. It was showed the lower melanogenesis rate on melanin production test by B16F10 cells as 27.46%. High inhibitory of tyrosinase was also measured as 28.61% by adding steamed Codonopsis lanceolata extracts by high pressure extraction of 1.0 mg/ml. And anti-wrinkle activity were 39.08%. In comparing phenolic acids profiles in the extract, in general higher amounts of polyphenol were obtained possibly by easy release of active components during thermal processing, which results in better antioxidant activities than that of general extract. This findings can also be supported by result that the extract by steaming process showed better activities than the general extraction extract.

According to previous reports, antioxidant activities of Codonopsis lanceolata could be increased by a steaming process. This study was performed to improve its antioxidant activity and skin whitening activities of C. lanceolata by high pressure and stepwise steaming complex process. The complex processed C. lanceolata showed highest free radical scavenging acitivity as 45.21%, and for phenol and flavonoid contents, complex processed C. lanceolata contained higher than those from conventional extraction process or steaming process alone. The Cytotoxicity of all C. lanceolata extracts also showed low cytotoxicity against human fibroblast cell (CCD-986sk) as 4.49 ~ 10.40%. In whitening activity, high inhibition of tyrosinase activity was estimated as 25.08% by adding the extracts from complex process. We found that whitening and antioxidant activity of complex processed C. lancolata extract was higher than those obtained from conventional extraction and a steaming process because various kinds of antioxidant compounds could be easily released by combined process, compared to one of each process.

We have established adventitious root culture systems of Codonopsis lanceolata, Codonopsis pilosula and Codonopsis ussuriensis. Root segments of C. lanceolata were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with 0.5mg/l NAA and produced 18.8±1.9 roots per explant. Root segments of C. pilosula were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with 1.0mg/l NAA and produced 8.5±1.8 roots per explant. Leaf segments of C. ussuriensis were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with 0.5mg/l NAA and produced 7.8±0.4 roots per explant. In liquid culture, the best production of adventitious root (fresh weight) was obtained in 1/2 MS medium with 1.0mg/l NAA. This study demonstrated for the first time to produce adventitious roots in C. pilosula and C. ussuriensis.

In this study, the changes that occurred in the quality of minimally processed sliced Deodeok (Codonopsis lanceolata) in relation to the packing method during storage at 7℃were investigated. The storage tests were conducted for seven days using PE sealing, but PP sealing and vacuum packaging preserved the Deodeok for 14 days. On the seventh day, the vinyl-packaged Deodeok showed a remarkable fall in quality with 4.5 °Brix, but the PP-sealed and vacuum-packaged Deodeok showed slight falls with 6.4 and 6.8 °Brix, respectively. The PE- and PP-sealed Deodeok did not show significant differences in texture and moisture content for two days, and the moisture content was highest in the vacuum-packaged Deodeok during storage. In relation to the total viable cell and the coliform count, the vacuum-packaged Deodeok showed the lowest rate of increment during storage, followed by the others. Thus, the bubble-washed and vacuum-packaged minimally processed sliced Deodeok was found to have the best quality.

In general, stepwise hot steaming process is known to be effective in improving its biological activities; however, not much employed in processing Codonopsis lanceolata due to its hardness. In this study, C. lanceolata was first pretreated with warm water at 50℃ and 60℃ for two hours, then steamed for 3 hours. Antioxidant activities of 70% ethanol extracts were compared with the extract from the water solvent: 41.58% vs 8.98% of DPPH radical scavenging activity in adding 1.25mg/ml of steamed extract and water extract, respectively. Reducing power of steamed and fresh C. lanceolata were also measured as 1.39 and 0.71. Total poly phenolic of the steamed extract was estimated as 12.11mg/g, compared to 3.98mg/g fresh C. lanceolata. Total flavonoid contents were also obtained as 11.48mg/g, compared to 7.11mg/g of fresh C. lanceolata. In comparing phenolic acids profiles in the extract, in general higher amounts of gallic acid, trans-ferulic acid, vanillic acid were obtained possibly by easy release of active components during thermal processing, which results in better antioxidant activities than that of water extract. This findings can also be supported by result that the ethanol extract showed better activities than the water extract.

The objectives of this study were to compare the antioxidant activities by high pressure extraction of Codonopsis lanceolata from different cultivation areas; Hoeng-sung, Jeju island, and China. Total phenolic acid contents of Hoeng-sung, Jeju, China were estimated as 732.11, 640.25, and 584.85 mg QUR/100 g DW, respectively. The flavonoids contents of Hoeng-sung, Jeju, China were measured as 80.37, 76.46, and 74.55 mg QUR/100g DW, respectively. Generally, contents of phenolic acid and flavonoids, HPE was higher than conventional extraction process. Hoeng-sung Codonopsis lanceolata showed 64.33% of DPPH radical scavenging activity (EDA, %) in 3.2 mg/ml of Hoeng-sung Codonopsis lanceolata. The reducing power of Hoeng-sung cultivation area Codonopsis lanceolata also showed the high activity as 3.15. In generally, antioxidant activities of Codonopsis lanceolata were increased by high pressure extraction process. Based on these results, higher contents of flavonoids and total polyphenols were found extracted by high pressure extraction of Codonopsis lanceolata grow in Hoeng-sung area than others.

The chromosome number was identified and fluorescence in situ hybridization(FISH) mapping of 5S and 45S rDNAs were conducted for C. minima and C. lanceolata in the genus Codonopsis from Jeju island. In this study, we have confirmed that the somatic metaphase chromosome number determined as 2n=2x=16 was the same as the findings from the previous studies. While the conventional staining method makes it rather difficult to distinguish satellite chromosomes due to high degree of variability, FISH analysis produced the exact number and location of 5S and 45S rDNAs. Both species in the genus Codonopsis have a pair of 5S rDNA and their gene loci were observed on chromosome 3. Although two pairs of 45S rDNAs (one on chromosome 1 and the other on chromosome 8) were identified in both species, the 45S rDNA signals on chromosome 8 in C. minima were significantly weaker than those on chromosome 1. In addition, the 45S rDNA signals on chromosome 1 in C. lanceolata showed that the chromosome is non-homologus. In this study, we have determined cytogenetic characteristics of C. minima and C. lanceolata according to their gene replication patterns.

Both fresh Codonopsis lanceolata and lactic acid bacteria fermented Codonopsis lanceolata were extracted with water at 100℃, and tested for anticancer activity using several human cancer cell lines. The fermented extracts inhibited the growth of hepatocellular carcinoma cells up to 90%, compared to 75% for fresh Codonopsis lanceolata. The extracts of cytotoxicity on human normal lung cells (HEK293) were as low as 15%. Especially, human hepatocellular carcinoma cell were more efficiently inhibited than other cells. This extract also inhibited α-glucosidase activity up to 60% at 1.0mg/ml. This fermented extracts showed the inhibition potency on tyrosinase by 25% at 1.0mg/ml. From the results, the fermented Codonopsis lanceolata enhanced several biological activities up to 20~30%, compared to those from fresh Codonopsis lanceolata. It implies that fermentation process could be one of useful methods of utilizing low quality Codonopsis lanceolata. Because this process could yield high amounts of biologically active compounds by the help of microbial growth.

Objectives The objective of our research was to establish the gene transformation, expression and characterization system in transgenic Codonopsis lanceolata. Materials and methods Agrobacterium tumefaciens strain LBA 4404/ binary vector pYBI121Regeneration of transgenic shoots: MS medium supplemented with 0.1 mg/ℓ NAA and 1 mg/ℓ BAP, 3% sucrose and 0.8% agar at pH 5.8. Agrobacterium cell density OD 600 between 0.8 and 1.0, Infection: 5 minutes DNA isolation and Polymerase chain reaction: DNA was extracted from young leafs excised from kanamycin resistant shoots. Two primers used for PCR amplification of the 700 bp of the npt II gene were N 1 (5′ GAA GCT ATT CGG CGG CTA TGA CTG 3′) as a sense primer and N 2 (5′ ATC GGG AGC GGC GGC GAT ACC CTA 3′) as a anti sense primer. Result and Discussion Adventitious shoots regenerated 3 weeks after Agrobacterium infection on regeneration medium containing 0.1 mg/ℓ NAA and 1 mg/ℓ BAP, 100 mg/ℓ kanamycin 250 mg/ℓ cefatoxime. Numerous adventitious shoot inductions of putative transformants were observed from the cut surface of explants which initially resembled knob like structure and later developed into new plant. PCR analysis of showed the expected bands of npt II gene. PCR analysis was carried out to confirm the insertion of the npt II gene in the genome of transformed plant. The expected amplified npt II fragments of size 700 bp was found in the T0 transformed plants, indicating the integration of npt II gene.

Field performance and morphological characterization was conducted on seven transgenic lines of Codonopsis lanceolata expressing γ-TMT gene. The shoots were obtained from leaf explants after co-cultivation with Agrobacterium tume-faciens strain LBA 4404 harboring a binary vector pYBI 121 that carried genes encoding γ-Tocopherol methyltransferase gene (γ-TMT) and a neomycin phosphotransferase II gene (npt II) for kanamycin resistance. The transgenic plants were transferred to a green house for acclimation. Integration of T-DNA into the T0 and T1 generation of transgenic Codonopsis lanceolata genome was confirmed by the polymerase chain reaction and southern blot analysis. The progenies of transgenic plants showed phenotypic differences within the different lines and with relative to control plants. When grown in field, the transgenic plants in general exhibited increased fertility, significant improvement in the shoot weight, root weight, shoot height and rachis length with relation to the control plants. However, all seven independently derived transgenic lines produced normal flower with respect to its shape, size, color and seeds number at its maturity. Indicating that the addition of a selectable marker gene in the plant genome does not effect on seed germination and agronomic performance of transgenic Codonopsis lanceolata. T1 progenies of these plants were obtained and evaluated together with control plant in a field experiment. Overall, the agronomic performance of T1 progenies of transgenic Codonopsis lanceolata showed superior to that of the seed derived non-transgenic plant. In this study, we report on the morphological variation and agronomic performance of transgenic Codonopsis lanceolata developed by Agrobacterium transformation.