'Green King', a new green tetraploid table grape cultivar with large berries, was originated from a cross between ‘Tensyu’ and ‘Beni Fuji’ grape cultivar in 2000. The cultivar was preliminarily selected among the elite breeding lines for its superior growth characteristics in 2005. After an extensive three-year evaluation of fruit and tree characteristics as 'GWG2002-02', 'Green King' was finally selected in 2008. Under Chuncheon’s ecological conditions, the average harvest date for 'Green King' is 6 September, and the number of days from flowering to harvest is 88 days. Hence, it is 12 days shorter than 'Rosario Bianco'. The average weight of a cluster in 'Green King' is 379.6 g, and its yield is 1,852 kg/10a. It has larger fruit size and higher fruit productivity than ‘Rosario Bianco’ (Registration No. 3560).

Background : Tetraploid plant is bigger in size of organs like leaf, stem, fruit, and root than diploid plant by chromosome doubling. Also, Self-ìncompatibílity disappeares and disease resistance is strengthened in tetraploid plant. To breed new boxthorn variety having bigger fruit, higher yield, pest tolerance, and, self-compatibílity, tetraploids was induced for tetraploid breeding resources by colchicine treatment in each boxthorn variety. Materials and Methods : Colchicine was treated in Myongan, Cheongdae, Bulro, and Youngha for 0.1% concentration and 48 hours. Polyploid was identified by flow cytomertry(Partec, PA-1) and chromosome numbers of polyploid individuals were examined by aceto-carmine stain. Conclusion : Colchicine was treated in Myongan, Cheongdae, Bulro, and Youngha for 0.1% concentration and 48 hours. Tetraploid was induced as 13, 9, 6, and 5 individuals in Myongan, Cheongdae, Bulro, and Youngha respectively by above colchicine treatment.

To produce high quality watermelon, three tetraploid watermelon breeding lines (‘SA03-1’, ‘SA06-1’ and ‘SB01-1’) were developed by treatment with different chromosome doubling reagents. To identify the optimal tetraploid inductive conditions, the three watermelon breeding lines were selected by counting the number of doubled chloroplasts in guard cells. Tetraploid induction rates differed depending on the genotypes and treatment with doubling reagents. However, the highest induction rate occurred with 1.0% colchicine (82.2%). These putative tetraploid lines were re-confirmed for ploidy using flow cytometric analysis and chromosome counting. The internode length of the tetraploid breeding lines was different when the leaf size was larger in all three tetraploid lines compared to their diploids. The fruit weight of the tetraploid fruits for ‘SA03-1’ and ‘SB01-1’ was lower than for their diploid, and the rind thickness and total sugar content (°Brix) of tetraploid SB01-1 were significantly different from those of its diploid. Tetraploid lines were sterile, yielded a lower number of seeds per fruit for ‘SA03-1’ (21), ‘SA06-1’ (62), and ‘SB01-1’ (34.7), and the seeds were larger and thicker than those of their diploids. These tetraploid breeding results will be useful for breeding new seedless watermelon cultivars.

본 연구에서는 4배체에서 염색체가 가감된 hypo-, hypertetraploid 포도들의 수체생육과 과실의 특성을 검정함으로서, 이들의 특성을 이용한 대립계 무핵포도 품종 육성 가능성에 대해서 검토하고자 하였다. Hypo-, hyper-tetraploid 포도들은 다른 이수체식물들에서 보고되었던 엽의 기형이나 수체생육의 불균일 현상 없이 4배체 포도인 ‘거봉’과 유사한 생육상을 보여 재배적으로 활용하는데 문제가 없는 것으로 나타났다. Hypo-teraploid 포도에서 만개기에 GA3 100 ppm 1회 처리 시 완전 무핵과 과실을 형성하였고, 과립도 8.5∼10.5 g 사이로 대립성 과실을 형성하였다. 하지만, Hyper-teraploid 포도는 hypo-teraploid 포도들에 비해 큰 과립을 가진 포도를 형성하였으나 낮은 무핵율을 나타나게 되는 것으로 조사되었다. 따라서 hypo-tetraploid 포도들의 특성을 이용하는 것은 대립계 무핵포도를 육성하기 위한 유용한 방법이 될 수 있을 것으로 판단된다.

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.

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.

Platycodon grandiflorum is a perennial flowering plant, known as Chinese bell flower, widespread in northeast Asia. The roots of this species are used for centuries to treat diseases, and have extensive pharmacological effects such as reducing adiposity, hyperlipidemia as well as anti-atherosclerotic disorder. In this study, systematical and targeting proteome analysis were executed from the 3, 4 and 5 months aged diploid and tetraploid roots of Platycodon grandiflorum and the proteins were separated by 2-DE and stained by CBB. In diploid roots, a total of 30 protein spots (≥ 2-fold) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Among the 30 differentially expressed proteins, 21 proteins sopts were identified as up-regulated and 9 proteins were identified as down-regulated. In contrary, a total of 40 differentially expressed proteins were confirmed from tetraploid roots whereas 28 protein spots were confirmed as up-regulated and 12 proteins were identified as down-regulated. However, the differentially expressed proteins from diploid and tetraploid roots were classified into 12 and 14 possible functional categories respectively using Protein Information Resources. The results revealed that the identified proteins from diploid and tetraploid roots were mainly involved in oxidoreductase activity, nucleotide binding, transferase activity and catalytic activity in bellflower roots. In conclusion, the exclusive proteins from diploid and tetraploid roots may provide insight clues for better understanding the characteristics and functions of proteins and metabolic activity of Platycodon grandiflorum.

Platycodon grandiflorum, known as Doraji in Korea, is used in various medications and traditional cuisine in Korea. This study was conducted to characterize the hormonal effects of diploid and tetraploid roots of P. grandiflorum using proteomics technique. Prior to proteome analysis, different kinds of growth hormones; IBA (1mg/L), NAA (1mg/L) and IAA (1mg/L) were applied in the adventitious (Diploid and tetraploid) roots for investigation. Solid (1/4MS) and liquid (1/2MS) medium were performed in the present study to investigate the hormonal effects. In diploid roots, two dimensional gels stained with CBB, a total of 1154 protein spots were identified using image analysis by Ludesi REDFIN 3 programme (Ludesi AB, Lund, Sweden: www.ludesi.com). Out of 1154 differential expressed protein spots, a total of 33 protein spots (≥ 2-fold) were selected for mass spectrometry. Among the 33 protein spots, 7 protein spots were up-regulated in IBA, 12 proteins in NAA and 14 proteins in IAA. In the case of tetraploid roots that performed under solid medium, a total of 842 differentially expressed protein spots were identified of which 34 proteins spots (≥ 1.5-fold) were selected for mass spectrometry. Out of 34 protein spots, 11 proteins were up-regulated in IBA, 10 proteins in NAA and 13proteins in IAA. However, a total of 659 differentially expressed proteins were confirmed from the liquid medium of tetraploid roots from which 32 proteins spots (≥ 1.5-fold) were sorted for MS analysis. Out of these 32 proteins, a total of 3 proteins were up-regulated in IBA, 7 proteins in NAA and 22 proteins in IAA. The identified proteins may provide insight clues for better understanding of the characteristics of proteins and biological activity from adventitious roots of Platycodon grandiflorum.

The antioxidant enzyme and DPPH radical scavenging activity with variations in drying methods of diploid and tetraploid in Platycodon grandiflorum were determined. Antioxidant enzyme activities were measured as superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), and ascorbate peroxidase (APX). The roots of Platycodon grandiflorum were freeze-dried, indoor-dried, hot-air dried, and microwave dried. The root extract of P. grandiflorum have shown the highest SOD enzyme activity of 92% in tetraploid of freeze-dried and indoor-dried while diploid of microwave dried showed the lowest SOD enzyme activity of 47.5%. The activity of CAT showed higher values in the root of tetraploid than in the diploid of P. grandiflorum in all drying methods. The APX activity showed relatively higher values in the root extract of freeze-dried both the diploid and tetraploid, but the difference in comparison with other extracts was not significant. The POX activities according to drying methods of diploid and tetraploid in P. grandiflorum showed relatively high values in freeze-dried and indoor-dried compared with other drying methods, and the POX activity between the diploid and tetraploid was not significant difference in each drying method. The DPPH radical scavenging activity with variation in drying methods of diploid and tetraploid in P. grandiflorum was the highest in the freeze-dried, and was higher in tetraploid than diploid in all the concentrations. In conclusion, the root of P. grandiflorum had the potent biological activities in both diploid and tetraploid. In particular, the tetraploid root of P. grandiflorum showing high antioxidant enzyme activity could be good materials for development of source of functional healthy food.

'망종화'에서 콜히친 처리에 의한 효율적인 4배체 식물을 유도하고자 적정 식물체 부위, 적정농도 및 침지시간을 검토하였다. 종자의 발아율은 콜히친의 농도가 높을수록 또한 침지시간이 길수록 저하되었다. 대조구를 제외한 16개의 처리구에서 총 453개체가 발아된 반면 4배체는 유도되지 않았다. 기내 배양 중인 줄기 절편체의 식물체 재생율은 콜히친 처리농도 0.01%에서 최고를 나타내다가 0.1% 농도 이상으로 높아질수록 낮았다. 적정식물체 부위는 줄기 절편체로 나타났다. 4배체 식물은 콜히친을 0.05% 이상으로 6시간 침지처리 하였을 때 얻을 수 있었고, 특히 0.05%, 12시간 침지처리에서 식물체의 재생수 대비 약 42%의 높은 획득율을 보였다. Flowcytometry에 의해 DNA함량의 배가여부를 확인한 결과, G1 phase의 DNA 함량 peak가 2배체에서 94.5, 4배체는 192.5로, DNA가 배가됨을 확인할 수 있었다. 또한 공변세포 당 엽록체 수는 2배체가 약 10개인 것에 비해 4배체는 17~19개로 2배체보다 약 1.7~1.9배 정도 많았다.

Aneuploidy breeding was utilized as a method to develop a new type ofdiploid Brassica crop. Novel combinations oftheA genome of Chinese cabbage and C genome of cabbage were established by crossing to produce a sesquidiploid and successivebackcrossing with

Present studies were carried out to produce tetraploid plants by colchicine treatment using seeds, seedlings and shoot tips of Codonopsis lanceolata. Three tetraploid plants of C. lanceolata were produced from seeds which absorbed 0.1 % colchicine solution for 12 hours, and 0.5% colchicine solution for 1 and 6 hours from seedlings, respectively. But tetraploid was not produced from shoot tips treated by colchicine solution. Compared to diploid, tetraploid plants had larger stomata, but less number of stomata. Fresh weight of tetraploid plants was 1.4∼3.6 times heavier than diploid plants.

Present studies were carried out to produce tetraploid plants by colchicine treatment using seeds, seedlings and shoot tips of Platycodon grandiflorum in Campanulaceae. The most successful colchicine treatment for tetraploid production in P. grandiflorum was soaking treatment using 0.01 and 0.5% colchicine solution for 1 hour and 12 hours, respectively. Morphological characteristics of both diploid and tetraploid were similar, but tetraploid plants had more leaves. Compared to diploid, tetraploid had the larger stomata, but less number of stomata. Fresh weight of tetraploids was 20∼40% heavier than that of diploid.

The standard protocol for the production of transgenic mouse from ES-injected embryo has to process via chimera producing and several times breeding steps, In contrast, tetraploid-ES cell complementation method allows the immediate generation of targeted murine mutants from genetically modified ES cell clones. The advantage of this advanced technique is a simple and efficient without chimeric intermediates. Recently, this method has been significantly improved through the discovery that ES cells derived from hybrid strains support the development of viable ES mice more efficiently than inbred ES cells do. Therefore, the objective of this study was to generate transgenic mice overexpressing human resistin gene by using tetrapioid-ES cell complementation method. Human resistin gene was amplified from human fetal liver cDNA library by PCR and cloned into pCR 2.1 TOPO T-vector and constructed in pCMV-Tag4C vector. Human resistin mammalian expression plasmid was transfected into D3-GL ES cells by lipofectamine 2000, and then after 8~10 days of transfection, the human resistin-expressing cells were selected with G418. In order to produce tetraploid embryos, blastomeres of diploid embryos at the two-cell stage were fused with two times of electric pulse using 60 V 30 sec. (fusion rate : 93.5%) and cultured upto the blastocyst stage (development rate : 94.6%). The 15~20 previously G418-selected ES cells were injected into tetraploid blastocysts, and then transferred into the uterus of E2.5d pseudopregnant recipient mice. To investigate the gestation progress, two El9.5d fetus were recovered by Casarean section and one fetus was confirmed to contain human resistin gene by genomic DNA-PCR. Therefore, this finding demonstrates that tetraploid-ES mouse technology can be considered as a useful tool to produce transgenic mouse for the rapid analysis of gene function in vivo.