Factors on Somatic Embryogenesis and Organogenesis of Soybean (Glycine max)

Kim, Kyong-Ho,Oh, Young-Jin,Suh, Sug-Kee,Kim, Hag-Sin,Kim, Tae-Soo,Lee, Mi-Ja,Park, Ho-Ki,Suh, Duck-Yong,Yeo, Up-Dong 한국콩연구회 2002 韓國콩硏究會誌 Vol.19 No.1

To enhance in vitro plantlet regeneration efficiency of soybean through embryogenesis, the culture conditions such as meterial part and size of immature seed, 2,4-D pH and solidifying agents for somatic embryogenesis were investigated. Somatic embryogenesis was induced from the immature embryo, immature cotyledon, and embryonic axis explants of the immature seed on MS medium supplemented with 2.0 mg/L 2,4-D. The highest rate (up tp 23%) of somatic embryogenesis was obtained from the immature cotyledon, followed by embryonic axis and immature embryo. The rate varied with the developmental stages of seed. The maximum rate (25%) of embryogenesis was obtained from 3-4 mm length of the seed after 25 days of flowering. The optimum concentration of 2,4-D for embryogenesis was 10mg/l. The optimum pH was at 5.8 and solidifying agent for medium was better with 0.4% gelrite than with agar. For rapid multiplication of shoot tip from germinating somatic embryos, they were cultured on MS medium containing 2 mg/L IBA and 1 mg/L BAP, After then somatic embryos with one and three cotyledons were transferred to the plant growth regulator free medium. The medium exhibited the higher rate (50%) of development than the multiplication medium. To select the section with shoot formation ability, the calli and shoot formation from three sections of 5-days-seedings of soybean were induced on MS medium supplemented with 1.0 mg/L BAP, 3% sucrose, and 0.3% gelrite for one month. The first leaf section exhibited the highest shoot formation rate (51%), followed the hypocotyl section and the cotyledon section. The shoot formation rates and shoot number of the four excised sections of the first leaf were aldo investigated on the same medium. A half of the first leaf explant and the third of the first leaf explant had higher shoot formation rates (76-80%) and number (3-4/ explants) than those in other two explants. Effects of cytokinins on shoot formation were determined, using the half of the first leaf explants. Zeatin (1.0mg/L) exhibited the highest in shoot formation rate (94%) and combination (1: 1, v/v) of zeatin (1.0mg/L) and IAA (1.0 mg/L) eshibited the highest in shoot formation rate (96%) and numbers (16/explant), twice more than zeatin alone (1.0 mg/L). The shoot cuttings were transferred and cultivated on the rooting media supplemented with only auxin, IBA at variois concentrations. The highest root formation (8/shoot) was achieved on the medium supplementes with 1.5 mg/L. After 4 weeks of cultivation, the plantlets with an extensive root system were transplanted in pots with a soil mixture of vermiculite and fine sand. Transferred to field, about 75% of the plantlets survived.

Recognition of Pharmacophore of ar-Turmerone for its Anticancer Activity

Baik, Kyong-Up,Jung, Sang-Hun,Ahn, Byung-Zun The Pharmaceutical Society of Korea 1993 Archives of Pharmacal Research Vol.16 No.3

For the evaluation of the role of .alpha., .betha.-unsaturated ketone portion of ar-tumerone for its activity, the structural variation of this structural unit was performed to omit its alkylating propery. Thus compounds 2, 3, and 4 were prepared and their cytotoxicities were determined against three different leukemiz cell lines (HL-60, K-562, and $L_{1210})$ in vitro. The biological inactivity against three different cell lines of these analogues implies that the $\alpha, \beta$-unsaturated ketone of ar-tumerone is the essential pharmacophore for its activity.

Structure Activity Relationship of ar-Turmerone Analogues

Baik, Kyong-Up,Jung, Sang-Hun,Ahn, Byung-Zun 충남대학교 약학대학 의약품개발연구소 1993 藥學論文集 Vol.9 No.-

For the analysis of structure activity relationship of ar-turmerone analogues, the compounds containing the various substituents on the phenyl ring and 1(or 2)-naphthyl group in the place of phenyl of ar-turmerone were prepared and tested their cytotoxicity against HL-60, K-562, and L1210 leukemia cells in vitro. The substituents at para position are methoxy, phenoxy, methyl, trifluoromethyl, fluoro, and chloro. At meta position methoxy, methyl, trifluoromethyl, or chloro groups and at ortho position methoxy or chloro group were introduced. Against HL-60 and K-562 cells, ED_50 values of the analogues are ranged from 0.8 to 30.0 ㎍/㎖. Against L1210 cell, these are located more than 20.0 ㎍/㎖. However, 5-carboethoxy-2-methyl-6-(1-naphthyl)-2-octen-4-one (5n) possesses ED50 valuses 0.8, 2.1, 6.5 ㎍/㎖ against HL-60, K-562, L1210 cells, respectively. The electronic nature of the subsituents on phenyl ring of ar-turmerone dose not affect the biological activity. Therefore the flat structure of aromatic portion of ar-turmerone analogues is the more important factor for their activity rather than its electronic nature. The potentiation of the cytotoxicity with the enlargement of aromatic ring region also supports the importance of the plane structure of this area. The restriction of the single bond rotation between C-6 and aromatic ring through the introduction of substituents at the ortho position of phenyl ring and the increment of size of alkyl group at C-6 position enhances the activity. Therefore the effective conformation should be the one having the orthogonal arrangement between the aromatic ring and the side chain.

Structure Activity Relationship of ar-Turmerone Analogues

Baik, Kyong-Up,Jung, Sang-Hun,Ahn, Byung-Zun The Pharmaceutical Society of Korea 1993 Archives of Pharmacal Research Vol.16 No.3

For the analysis of structure relationship of ar-turmerone analogues, the compounds containing the various substituents on the phenyl ring and 1(or 2)-naphthyl group in the place of phenyl of ar-turmerone were prepared and tested their cytotoxicity against HL-60, K-562, and L1210 leukemia cells in vitro. The substituents at para position are methoxy, phenoxy, methyl, trifluoromethyl, fluoro, and chloro. At meta position methoxy, methyl, trifluoromethyl, or chloro groups at ortho position mathoxy or chloro group were introduced. Against HL-60 and K-562 cells, $ED_{50}$ values of the analogues are ranged from 0.8 to $30.0\;\mu{g/ml}$. Againste L1210 cell, these are located more than $20.0\;\mu{g/ml}$. However, 5-carbone-thoxy-2-methyl-6(1-naphthyl)-2-octen-4-one (5n)possesses $ED_{50}$ valuses 0.8, 2.1, $6.5\;\mu{g/ml}$ against HL-60, L1210 cells, respectively. The electronic nature of the substituents on phenyl ring of ar-tumerone dose not affect the biological activity. Therefore the flat structure of aromatic potion of ar-tumerone analogues is the more important factor for their activity rather than its electronic nature. The potentiation of the cytotoxicity with the enlargement of aromatic ring region also supports the importance of the plane structure of this area. The restriction of the single bond rotation between C-6 and aromatic ring through the introduction of substituents at the ortho position of phenyl ring and the increment of size of alkyl group at C-6 position enhances the activity. Therefore the effective conformation should by the one having the orthogonal arrangement between the aromatic ring and the side chain.

The Factors on Somatic Embryogenesis of Soybean [Glycine max. (L.) Merrill]

Kim, Kyong-Ho,Kim, Hag-Sin,Oh, Young-Jin,Suh, Sug-Kee,Kim, Tae-Soo,Park, Ho-Kee,Park, Moon-Soo,Kim, Seok-Dong,Yeo, Up-Dong The Korean Society of Plant Biotechnology 2000 Plant molecular biology and biotechnology research Vol.2 No.3

To enhance in vitro plantlet regeneration efficiency of soybean through embryogenesis, the culture conditions such as material part and size of immature seed, 2,4-D, pH and solidifying agents for somatic embryogenesis were investigated. Somatic embryogenesis was induced from the immature embryo, immature cotyledon and embryonic axis explants of the immature seed on MS medium supplemented with 2.0 mg/L 2,4-D. The highest rate (up to 22.9%) of somatic embryogenesis was obtained from the immature cotyledon, following embryonic axis and the immature embryo. The rate varied with the developmental stages of seed. The maximum rate (25.4%) of embryogenesis was obtained from 3-4 mm length of the seed (after 25 days of flowering). The optimum concentration of 2,4-D for embryogenesis was 10 mg/L. The optimum pH was at 5.8 and solidifying agent for medium was better with 0.4% gelrite than with agar. For rapid multiplication of shoot tips from the germinating somatic embryos, they were cultured on MS medium containing 2 mg/L indole-3-butyyic acid (IBA) and 1 mg/L 6-benzyladenine (BA). After then somatic embryos with one and three cotyledons were transferred to the growth regulator free medium. The medium exhibited the higher rate (ca. 50%) of development than the multiplication medium.

Recognition of Pharmacophore of ar-Turmerone for its Anticancer Activity

Baik, Kyong-Up,Jung, Sang-Hun,Ahn, Byung-Zun 충남대학교 약학대학 의약품개발연구소 1993 藥學論文集 Vol.9 No.-

For the evaluation of the role of α,β-unsaturated ketone portion of ar-turmerone for its activity, the structural variation of this structural unit was performed to omit its alkylating property. Thus compounds 2, 3, and 4 were prepared and their cytotoxicities were determined against three different leukemia cell lines (HL-60, K-562, and L_1210) in vitro. The biological inactivity against three different cell lines of these analogues implies that the α,β-unsaturated ketone of ar-turmerone is the essential pharmacophore for its activity.

Changes in Cell-Wall Polysaccharide Contents during Growth of Suspension-Cultured DCB-Habituated Cells of Tobacco (Nicotiana tabacum L. cv. BY2)

Kyong Ho Kim,Up Dong Yeo,Nakagawa Naoki,Sakurai Naoki 한국육종학회 2001 한국육종학회 학술발표회 발표요지 Vol.2001 No.9

담배 ( Nicotiana glauca Graham ) Callus 배양시 Nicotine 생성에 미치는 2 , 4-D 및 NAA 의 영향

여읍동(Up Dong Yeo),김경호(Kyong Ho Kim) 한국식물학회 1991 Journal of Plant Biology Vol.34 No.2

Effects of two auxins, 2,4-dichlorophenoxyacetic acid (2,4-D) and α-napthaleneacetic acid (NAA) on nicotine production during callus culture of a wild tobacco (Nicotiana glauca) were investigated using a high performance liquid chromatography (HPLC). The high concentration (11.5 μM) of 2,4-D and NAA had peaks of nicotine contents at 4th and 2nd week, respectively. Thereafter, the concents decreased and the nicotine was metabolized to other alkaloids, The low concentration (1.5 μM) of 2,4-D on the medium supplemented with 0.1 mM of L-aspartic acid of L-arginine inhibited nicotine production. However, the low NAA promoted it only when the medium was supplemented with L-aspartic acid. From these results, it could be concluded that both auxins exhibit different action mechanisms on nicotine production pathway and the low NAA promotes the activities for the pathway with L-aspartic acid as a precursor.

Long-Term Effects of Growth Regulators on Growth and Turnover of Symplastic and Apoplastic Sugars in the Suspension Subculture of Kidney Bean

Yeo, Up-Dong,Pandey, Dev Mani,Kim, Kyong-Ho 한국식물학회 2004 Journal of Plant Biology Vol.47 No.1

Suspension cells of kidney bean were grown for 42 d in MS medium supplemented with growth regulators (2.0 mgL^(-1) 2,4-D and 0.5 ㎎L^(-1) kinetin) or without. At the stationary growth phase (42 d), the sugars were fractionated into the symplastic (ethanol and starch) and apoplastic [low-molecular pectin (Im-pectin), high-molecular pectin (hm-pectin), hemicellulose, and cellulose] sugars. The neutral sugars (NS) of hm-pectin and hemicellulose fractions were analyzed by GLC. The growth of the suspension cells in the liquid MS media, in terms of settled cell volume (SCV), remained similar, to the end of the experiment, irrespective of the presence or absence of growth regulators, indicating the nonnecessity of the exogenous growth regulators for the subculture. Total sugar(TS) of the ethanol fraction and NS of the Im-pectin of the suspension cells grown in the medium with growth regulators were higher than in the medium without growth regulators. However, starch content in the starch fraction and uronic acid (UA) content of the Im-pectin fraction did not exhibit any differences. From these results, it was suggested that the growth regulators modulated the structure of the cell wall polysaccharide. Analysis of the NS composition of the Im-pectin fractions revealed that the Rha, Arb, and Gal contents in the presence of growth regulators were higher than in the absence, while the Xyl, Man, and Glc contents in the presence of growth regulators were higher than in the absence, indicating the turnovers of rhamnogalacturonan and/or arabinogalactan. On the other hand, analysis of NS composition of hemicellulose fractions revealed that the Ara and Glc contents in the presence of growth regulators was higher than in the absence, whereas Xyl and Glc contents were nearly consistent, indicating the turnovers of arabinaogalactan Ⅰ or Ⅱ. The cellu- lose contents remained similar, irrespective of the presence (19.1%) or absence (18.7%) of growth regulators.

Effects of Auxins and Cytokinins on Organogenesis of Soybean Glycine max L.

Kim, Kyong-Ho,Park, Ho-Ki,Park, Moon-Soo,Yeo, Up-Dong The Korean Society of Plant Biotechnology 2001 Plant molecular biology and biotechnology research Vol.3 No.2

To select the section with shoot formation ability, the calli and shoot formation from three sections (first leaf including cotyledonary node, hypocotyl and cotyledon explants) of 5-days-seedlings of soybean were induced on MS medium supplemented with 1.0 mg/L BAP, 3% sucrose, and 0.3% gelrite for one month. The first leaf section exhibited the highest shoot formation rate (51%), followed the hypocotyl section (10%) and the cotyledon section (0%). The shoot formation rates and shoot number of the four excised sections (whole first leaf, a half of the first leaf, a third of the first leaf and only node) of the first leaf were also investigated on the same medium. A half of the first leaf explant and the third of the first leaf explant had higher shoot formation rates (76-80%) and numbers (3-4 / explants) than those in other two explants. Effects of six cytokinins (kinetin, zeatin, BAP, 2iP, PBA, and TDZ) on shoot formation were determined, using the half of the first leaf explants. Zeatin (1.0 mg/L) exhibited the highest in shoot formation rate (94%) and numbers (8 / explant). In addition, the combined effects of three cytokinins (zeatin, BAP, and TDZ; 0.5, 1.0, 2.0 mg/L, respectively) and an auxin (IAA; 0.0, 0.5, 1.0, 2.0 mg/L) were determined. The combination (1:1, v/v) of zeatin (1.0 mg/L) and IAA (1.0 mg/L) exhibited the highest in shoot formation rate (96%) and numbers (16 / explant), twice more than zeatin (1.0 mg/L) alone. The shoot cuttings were transferred and cultivated on the rooting media supplemented with only auxin, IBA at various concentrations. The highest root formation (8 / shoot) was achieved on the medium supplemented with 1.5 mg/L. After 4 weeks of cultivation, the plantlets with an extensive root system were transplanted in pots with a soil mixture of vermiculite and fine sand. Transferred to field, about 75% of the plantlets survived.