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Comparative Antioxidant Enzyme Activity of Diploid and Tetraploid Platycodon grandiflorum by Different Drying Methods

  • Boo, Hee-Ock;Shin, Jeoung-Hwa;Kim, Young-Seon;Park, Hee-Jeong;Kim, Hag-Hyun;Kwon, Soo-Jeong;Woo, Sun-Hee
    • Korean Journal of Plant Resources
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    • v.26 no.3
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    • pp.389-396
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    • 2013
  • 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.

Medium compositions reveal potential organogenesis in the diploid and tetrploid Codonopsis lanceolata

  • Kwon, Soo Jeong;Hwang, Ha Nule;Moon, Young Ja;Cho, Gab Yeon;Boo, Hee Ock;Lee, Moon Soon;Woo, Sun Hee;Kim, Hag Hyun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.169-169
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    • 2017
  • Medium composition plays a key role on influencing organogenesis in plant tissue culture. This study was carried out to examine the effects of medium composition on organogenesis in diploid and tetraploid Codonopsis lanceolata and obtain in-vitro mass propagation of superior species of C. lanceolata. Diploid C. lanceolata was found to be declined regarding MS medium composition for each concentration. However, shoot and adventitious root formation were suppressed with higher mineral salt concentration, and active growth of shoot and adventitious root was exhibited as 4.9 cm and 3.2 cm respectively in 1/2 MS medium. While in tetraploid C. lanceolata, it showed 2.9 cm and 3.2 cm respectively in 1/4 MS medium. In the case of sucrose concentration, no consistent decrease was observed for growth of shoot and the adventitious root of diploid both at high and low concentration. The growth of shoot (at 3% concentration) and adventitious root (at 7% concentration) was 2.3 cm and 2.0 cm respectively. Although there was no difference in shoot formation of tetraploid C. lanceolata in all concentrations with the range of 1.7~1.8, there was a slight decrease in shoot growth at high concentration. Results revealed that the adventitious root formation was suppressed at high concentration. The concentration of agar exhibited no significant difference in shoot formation of diploid C. lanceolata at all concentrations. The maximum result of adventitious growth (4.1 cm) was observed at 0.8% concentration. Slight inhibition of shoot formation and root formation of tetraploid C. lanceolata was observed at higher concentration. Shoot formation of diploid C. lanceolata also exhibited inhibition at higher concentration. Shoot formation of diploid C. lanceolata was increased at lower pH and shoot growth was the highest (2.3 cm) at pH 3.8. Adventitious root formation was higher at lower pH. However, both the adventitious root formation and growth exhibited comparatively higher result at pH 5.8. Taken together, the levels of pH had an effect on shoot and root formation in diploid and tetraploid of C. lanceolata

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Effect of Growth Regulators on Organogenesis in Diploid and Tetraploid Codonopsis Ianceolata In Vitro Culture

  • Soo-Jeong Kwon;Young-Ho Choi;Hee-Ock Boo;Sun-Hee Woo;Hag-Hyun Kim
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.161-161
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    • 2022
  • This study aimed to investigate the effect of growth regulators on the formation of the organ in the in vitro propagation of diploid and tetraploid Codonopsis lanceolata, and gain the basic data for in vitro propagation of superior C. lanceolata. In the case of diploid C. lanceolata, the highestshoot formation (3.0) was observed at 0.5 mg·L-1 addition medium with low IBA concentration. The shoot formation of tetraploid C. lanceolata was suppressed by addition of IBA. In the addition of lAA, the shoot formation of diploid C. lanceolata was slightly higher at 1.0 mg·-L-1 addition medium than that of control group, whereas tetraploid C. lanceolata showed the highest number (5.4) from control group. In the case of NAA, the shoot formation of diploid and tetra C. lanceolata tended to decrease at higher concentration. In terms of BA addition, the shoot formation of diploid C. lanceolata was increased by the addition of BA, whereaswhile the growth of shoot was decreased by the addition of BA. In the case of tetraploid C. lanceolata, shoot was found to be formed by the addition of low concentration of BA, and the growth of shoot was inhibited with the higher addition concentration of BA. With the addition of kinetin, the shoot formation of diploid C. lanceolata was slightly higher than that of control group, and the formation of adventitious root was highest (5.3) in the control group. In the case of tetraploid C. lanceolata, the shoot formation was similar in all treatment groups, but the formation and growth of adventitious root were significantly lower than that of diploid C. lanceolata. In the case of TDZ addition, the shoot formation of diploid C. lanceolata showed the pronounced results at 5.0 mg·L-1 addition medium, and the growth of shoot was inhibited by the addition of TDZ. The formation of adventitious root was 5.3 and 4.9 in the control group and 0.1 mg·L-1 addition medium respectively. The formation of the shoot of tetraploid C. lanceolata showed better results with the higher concentration of TDZ, and the growth was better with the lower concentration of TDZ. The formation and growth of adventitious root were significantly slower than that of diploid C. lanceolata.

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Evaluation of Cytotoxicity, Antimicrobial and Antioxidant Enzyme Activity of Diploid and Tetraploid Platycodon grandiflorum

  • Boo, Hee-Ock;Kim, Young-Sun;Kim, Hag-Hyun;Kwon, Soo-Jeong;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.2
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    • pp.239-247
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    • 2015
  • 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 $IC_{50}$ (concentration causing 50% cell death) value showed the highest on HCT-116 cell ($105.91{\mu}g/mL$), and exhibited significant activity against the Hep 3B cell ($140.67{\mu}g/mL$), SNU-1066 cell ($154.01{\mu}g/mL$), Hela cell ($158.37{\mu}g/mL$), SNU-601 cell ($182.67{\mu}g/mL$), Calu-6 cell ($190.42{\mu}g/mL$), MCF-7 cell ($510.19{\mu}g/mL$). 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.

Comparative proteome analysis of diploid and tetraploid root in Platycodon grandiflorum

  • Kwon, Soo Jeong;Roy, Swapan Kumar;Yoo, Jang-Hawan;Cho, Seong-Woo;Kim, Hag Hyun;Boo, Hee Ock;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.123-123
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    • 2017
  • In spite of the potential medicinal significance and a wide range of pharmacologic properties of Platycodon grandiflorum, the molecular mechanism of its roots is still unknown. The present study was conducted to profile proteins from 3, 4 and 5 months aged diploid and tetraploid roots of Platycodon grandiflorum using high throughput proteome approach. Two-dimensional gels stained with CBB, a total of 68 differential expressed proteins were identified from the 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 (${\geq}2-fold$) 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 (${\geq}2-fold$) 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. Taken together, the identified proteins may be helpful to identify key candidate proteins for genetic improvement of plants.

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Proteome characterization of hormone-induced diploid and tetraploid roots of Platycodon grandiflorum

  • Kwon, Soo Jeong;Roy, Swapan Kumar;Cho, Seong-Woo;Kim, Hag Hyun;Boo, Hee Ock;Song, Beom-Heon;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.132-132
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    • 2017
  • Plants, including Platycodon grandiflorum have been used globally across varied cultures as a safe natural source of medicines. From time immemorial, humans have relied on plants that could meet their basic necessities such as food, shelter, fuel and health. This study was executed 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 ( ${\geq}1.5-fold$) were analyzed using LTQ-FTICR MS 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 (${\geq}1.5-fold$) 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 protein function and its metabolic activity that can help for the development of the nutritional and breeding aspects of this economically important medicinal plant.

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Response of colchicine for the efficient chromosome doubling in Codonopsis lanceolata

  • Kwon, Soo-Jeong;Lee, Ui Gun;Moon, Young Ja;Cho, Gab Yeon;Boo, Hee Ock;Lee, Moon Soon;Woo, Sun Hee;Kim, Hag Hyun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.273-273
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    • 2017
  • Polyploidy has opened a new horizon for selection to sculpt a variety of new gene functions, traits, and lineages. The aim of this study was to investigate the efficacy of the colchicine concentration, temporal changes, and suitable material for inducing effective tetraploid plants of Codonopsis lanceolata. A total of 180 individuals from 16 treatment groups were germinated, and exposed to different concentrations of Colchicine. The plant height of the diploid (18.1 cm) was slightly shorter than that of the tetraploid (13.4 cm). The fresh weight of the main root in the diploid (0.5 g) was 4-fold higher than the tetraploid (2.2 g). The colchicine-treated plant regeneration rate in C. lanceolata was decreased at the elevated concentration of colchicine. A total of 126 individual plants were regenerated in the entire treatment group and tetraploid (2n=4x=32) plants were obtained. In particular, 5 individuals of the tetraploid plant were induced in the 0.05% colchicine for 6h, which is a higher rate (29.4%) than other regenerated plants. As in the seed treatment result, the plant height of the diploid was significantly higher (10.4 cm) than tetraploid. The root length of the tetraploid (10.1 cm) was longer than the diploid, and the root was also thicker. Taken together, the results obtained from the present study may be helpful for the efficient recovery of such polyploid plants through the in vitro application of colchicine, and may improve the productivity and breeding of C. lanceolata.

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Effect of NaCI Concentration on Germination and Seedling Growth of Italian Ryegrass(Lolium multiflorum L.) (NaCl 농도가 이탈리안 라이그래스의 발아와 유묘생장에 미치는 영향)

  • 이강수;최선영;최철원
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.3
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    • pp.340-350
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    • 1995
  • This study was conducted to obtain the basic information for the salt stress of Italian Ryegrass. Fourteen cultivars including 7 diploid and 7 tetraploid were subjected to eight levels of NaCl treatments during germination and early seedling stages. Germination percentage decreased remarkably over 300mM of NaCl concentration. The NaCl concentration that inhibited germination of 50% of the viable seeds in the cultivars was 344mM. The decreasing degree of germination percentage was higher in tetraploid type than in diploid type. The varietal difference of germination percentage was shown to be very conspicuous as the NaCl concentration increased. Top dry weight in 166 mM NaCl was decreased by 50% compared with that of control. Inhibition of NaCl appeared to be more in early seedling stage than in germination stage. Top dry weight of cultivars in NaCl stress was not related with germination capacity. Root dry weight in 148mM NaCl decreased by 50% compared with that of control. Root dry weight of cultivars was significantly correlated with top dry weight in NaCl treatment. Top dry weight and root dry weight was higher in diploid type than in tetraploid type at each 8 levels of NaCl concentration. Total chlorophyll content increased with the increase of NaCl concentration and the degree of increase in chlorophyll a was higher than that in chlorophyll b. Chlorophyll a was higher in tetraploid type than in diploid type. But chlorophyll b was higher in diploid type than in tetraploid type. There were significant correlation between chlorophyll a and top dry weight of cultivars over 150mM of NaCl concentration. Free proline content accumulated remarkably in NaCl treatment compared with control and was not difference between diploid type and tetraploid type. Free proline content of cultivars was not related with top dry weight in NaCl treatment.

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Breeding of Tetraploid in Platycodon grandiflorum (Jacq.)A. DC. by Colchicine treatment

  • Kim, Ik-Hwan;Kim, Hag-Hyun;Hong, Eui-Yon;Yun, Jong-Sun;Yun, Tae;Hwang, Ju-Kwang;Lee, Cheol-Hee
    • Plant Resources
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    • v.6 no.3
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    • pp.188-194
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    • 2003
  • 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.

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Proteomic Responses of Diploid and Tetraploid Roots in Platycodon grandiflorum (2배체와 4배체 도라지의 단백질 발현양상 비교 분석)

  • Kim, Hye-Rim;Kwon, Soo-Jeong;Roy, Swapan Kumar;Cho, Seong-Woo;Kim, Hag-Hyun;Moon, Young-Ja;Boo, Hee-Ock;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.3
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    • pp.394-400
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    • 2015
  • 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 (${\geq}\;1.5$-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.