• Title/Summary/Keyword: Isozyme pattern

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Improvement of Peroxidase Productivity by Optimization of Medium Composition and Cell Inoculum Size in Suspension Cultures of Sweet Potato (Ipomoea batatas) (고구마(Ipomoea batatase)현탁배양에서 배지조성 및 세포접종량의 적정화에 의한 Pemxidase생산성 향상)

  • 곽상수;김수경;정경희;유순희;박일현;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.2
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    • pp.91-97
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    • 1994
  • To improve the productivity of peroxidase (POD) of cell line SP-47 derived from cell suspension cultures of sweet potato (Ipomoea batatas (L) Lam.cv White Star), we optimized culture conditions including the composition and concentration of plant growth regulators and carbon source, and the cell inoculum size. When one g (fr wt) of cells was inoculated into 50 mL TL medium supplemented with l mg/L 2,4-D and 30g/L sucrose in 300 mL Erlenmeyer flask at 25$^{\circ}C$ in the dark (100rpm), the POD activity per g cell dry wt was maximized to be about 6,800 units after 25 days of subculture, which was about 30 times higher than that of intact roots of horseradish plants grown in the greenhouse, but the cell growth was maximum after 15 days of subculture. The protein content per g cell dry wt maintained almost plateau and after 25 days of subculture decreased as culture Proceeded further whereas the POD specific activity (unit/mg protein) was about two times higher after subculture and continuously increased from 12 days to the end of cultures (40 days). The POD isozyme patterns showed almost the same regardless of cell growth stage, but some acidic isozymes were slightly increased after 25 days of subculture. These results indicate that POD activity in suspension cultures of sweet potato is closely associated with cell growth and stresses derived from cell culture renditions and medium depletion. Due to its high POD activity the SPL47cell line seems to be suitable for the mass production of POD.

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Superoxide Dismutase and Peroxidase Activity of Transformed Callus in Tomato (형질전환된 토마토 캘러스의 Superoxide Dismutase와 Peroxidase 활성)

  • 유정민;정형진;김경민;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.177-181
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    • 1998
  • This study was carried out to investigate activity difference in the superoxide dismutase (SOD) and peroxidase (POD) of tomato callus transformed with Agrobacterium containing the GUS gene. Than those of other two tomato cultivars, the hypocotyl explant of JA101 was shown to have higher POD and SOD specific activity of 23 unit/mg protein and 2,156 unit/mg protein, respectively. Relatively high frequency of callus formation was obtained from the hypocotyl explant on MS medium containing 1 mg/L 2,4-D for 30 days and its POD(47 unit/mg protein) and SOD (95,786 unit/mg protein) specific activities were higher than other 2,4-D concentration. The hypocotyl explant and callus cocultivated with Agrobacterium for 72 hours were transferred to MS medium supplemented with 1 mg/L 2,4-D, 30 mg/L kanamycin, 30 g/L sucrose and 4 g/L Gelrite. The hypocotyl explants transferred to the medium formed callus with 45.5% effeciency after 8 weeks. The transformation efficiency confirmed by GUS assay was 21.6%. POD specific activity of the transformed callus (54 unit/mg protein) were somewhat lower than the non-transformed callus (64 unit/mgg protein) and SOD specific activity of the transformed callus (30,300 unit/mg protein) were also lower than the non-transformed callus (37,077 unit/mg protein). However there was no significant difference in POD and SOD isozyme patterns between the transformed and the non-transformed calluses. From these results, it revealed that there was no difference of antioxidant enzyme activities between the transformed callus and the non-transformed callus in tomato.

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A Study on electrophoresis and Activity of Alcohol Dehydrogenase of Drosophila melanogaster (한국산 노랑초파리 (Drosophila melanogaster) Alcohol dehydrogenase의 전기영동산 및 활성에 대한 연구)

  • Chung, Yong-Jai;Yoon, Youn-Sik
    • The Korean Journal of Zoology
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    • v.23 no.3
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    • pp.125-136
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    • 1980
  • The purpose of the present study is to investigate the electrophoretic patterns of ADH isozymes and activity at various developmental stages of Oregon-R and Kwangju strains of D. melanogaster. The ADH isozymes were seperated by agarose gel plate eletrophoresis and they were stained for ADH activity study. The optical density was measured by using densitometer and the total activity was determined by measuring the enzyme solution's absorbance of 340 nm by spectrophotometer. The results are as follows: 1. At egg stages of Oregon-R and Kwangju strains ADH 2, ADH 3, ADH 4 were observed in te zymograms, but all of ADH 1, ADH 2, ADH 3, ADH 4 and ADH 5 were detected in larval, pupal and imaginal stages. 2. The amount of ADH 1+ADH 2 (anion) was 20-40% and that of ADH 3+ADH 4+ADH 5 (cation) was 60-805. In the developmental stages other, than egg stages, ADH 3 of both strains showed the largest amount among all ADH isozymes, and especially ADH 4 of Kwangju strain showed less amount than that of Oregon-R strain. 3. The total activity of the ADH at the egg stage was found to be the lowest, at larval stage became to be higher and to be lower at pupal stage and then became to be the highest at the imaginal stage. It is noticeable that the activity of the ADH in Kwangju strain was higher than that in Oregon-R strain.

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Effect of Shading Practices on the Chemical Compounds and Antioxidant in Aruncus dioicus (차광재배가 눈개승마(Aruncus dioicus var. kamtschaticus)의 성분 및 항산화 특성에 미치는 영향)

  • Kwon Jeong-Woo;Park Jae-Ho;Kwon Ki-Soo;Kim Dae-Seup;Jeong Jin-Boo;Lee Hee-Kyung;Sim Young-Eun;Kim Mi-Suk;Youn Ji-Young;Chung Gyu-Young;Jeong Hyung-Jin
    • Korean Journal of Plant Resources
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    • v.19 no.1
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    • pp.1-7
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    • 2006
  • This study was conducted to determine chemical compounds and antioxidant in Aruncus dioicus. Further, effects of cultural practices such as the shading conditions on the general compounds, essential oils and antioxidants. The chemical compounds and antioxidant activity of edible extracts on the shading cultivation and harvesting time were measured by crude fiber, pigments, higher fatty acids, essential oils and DPPH free radical scavenging ability and activities of SOD and POD in edible shooting parts of Aruncus dioicus. The contents of crude fiber, total chlorophyll and total carotenoids in extracts of edible shooting by shading cultivation and later harvesting were higher than those of non-shading cultivation and early harvesting. Phenolic compounds such as benzoic acid were identified as the aromatic compounds in the edible extracts of Aruncus dioicus. The contents of azulene, benzaldehyde and linalool among those compounds on the cultivation conditions increased in shading cultivation. $IC_{50}$ values of DPPH free radical scavenging activities were high from 6.644 to $14.499{mu}g/m{\ell}$ in extracts of edible parts and those of edible shooting parts was high such as edible shooting cultivated by 60% shading, 30% shading and non-shading, respectively. The activity of POD and SOD in seeds was lower than that of edible shooting and that by shading cultivation was high in extracts cultivated by non-shading. POD activity of extracts on harvesting time was high in earlier harvesting but SOD activity was low. The numbers of isozyme pattern of POD and SOD in seedling showed 7 bands and 3 bands, respectively, especially, bands of POD and SOD in the first year-growing plant did not show and show a difference according to plant positions, respectively.

Purification of Mitochondrial Matrix Aldehyde Dehydrogenase from Pig Brain

  • Kim, Kyu-Tae;Lee, Young-Don
    • BMB Reports
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    • v.28 no.2
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    • pp.177-183
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    • 1995
  • The activity of aldehyde dehydrogenase (ALDH) in the cerebrum, cerebellum, striatum, and medulla oblongata was examined and mitochondrial matrix ALDH was purified prior to immunohistochemical study on the localization of ALDH isozymes in pig brain. Relatively high enzyme activity was found in the striatum and medulla oblongata when using indole-3-acetaldehyde as substrate, and in the striatum when using 3,4-dihydroxyphenylacetaldehyde (DOPAL). The main part of mitochondrial ALDH activities with both acetaldehyde and DOPAL existed in the matrix fraction. The ratio of activity of the matrix to the membrane fraction in the cerebrum was higher than in the cerebellum, suggesting that the distribution pattern of ALDH isozymes was different according to the brain regions. The 276-fold purified mitochondrial matrix ALDH from pig brain was identified to be homologous tetramers with 53 KD subunits. The enzyme showed maximal activity at pH 9.0 and was stable in the temperature range from $25^{\circ}C$ to $37^{\circ}C$. The mitochondrial matrix ALDH activity was considerably inhibited by acetaldehyde in vitro. The $K_m$ values of the enzyme for acetaldehyde and propionaldehyde were 5.8 mM and 4.9 mM, respectively, whereas $K_m$ values for indole-3-acetaldehyde and DOPAL were 44 ${\mu}M$ and 1.6 ${\mu}M$, respectively. The $V_{max}/K_{m}$ ratio was the highest with DOPAL as compared with other substrates. These results suggested that mitochondrial matrix ALDH in the present work might be a low Km isozyme involved in biogenic aldehyde oxidation in pig brain.

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Influence of Hexaconazole on Biochemical Constituents of Groundnut (땅콩의 생화학성분에 미치는 Hexaconazole의 영향)

  • Johnson, I.;Marimuthu, T.;Samiyappan, R.;Cha, Byeong-Jin
    • The Korean Journal of Pesticide Science
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    • v.12 no.4
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    • pp.335-341
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    • 2008
  • In this study, hexaconazole 5% SC, an ergosterol biosynthesis inhibitor, was tested on groundnut with its recommended ($500\;mL\;ha^{-1}$) and higher ($2,000\;mL\;ha^{-1}$) concentrations under greenhouse conditions in India. Its influence on biochemical constituents of groundnut plants was assessed apart from its disease management potential against late leaf spot caused by Phaeoisariopsis personata (Berk and Curt). Likewise, leaf samples were collected from hexaconazole 5% SC-sprayed plants at different time intervals. Thereafter, their analyses showed considerable differences in the plant constituents, such as chlorophyll, soluble protein, and total phenol contents and the activity of nitrate reductase enzyme. The induction activity of defense-related enzyme, peroxidase, was also analyzed. However, no difference was observed in the isozymic pattern. Moreover, the ground kernels collected from treated plants also showed no difference in the estimated carbohydrate and other constituents.

Breeding of the native vegetables using the biotechnology

  • Iwamoto, Yuzuri
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.106-111
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    • 2005
  • For breeding of a new rootstock for eggplant production, somatic hybrids between two species, Solanum integrifolium and S. sanitwongsei were obtained through protoplast fusion. The former species has been commonly used for rootstock for eggplant production in Japan. Eggplants on these rootstocks are more productive than ungrafted plants, but are susceptible to bacterial wilt caused Ralstonia solanacearum. While the latter species is resistant, the growth of eggplants on this rootstock is rather slow and low yield. Protoplast of both species were isolated from cotyledons, and inactivated with iodoacetamide or UV-irradiation, then fused electrically. The fused products were then cultured. Regenerated plantlets were then transplanted on soil then maintained in a green house. The plants were classified into four groups. Those in the first group showed morphological characters intermediate of the parentalspecies. The plants bore fruit with viable seeds. The plants showed a chromosome number of 2n=48, the sum of those of the parental species, and are suggested to be symmetric fusion products. While plants in the other groupswas less vigorous and showed chromosome number 2n= 68 to 72 suggesting asymmetric fusion products by genomic in situ hybridization(GISH). Isozyme pattern of shikimate dehydrogenase (SKDH; EC 1.1.1.25), isocitrate dehydrogenase (IDH; EC 1.1.1.41) and phosphoglucomutase (PGM; EC 2.7.5.1) showed that 24 regenerated plants in three groups were somatic hybrids. Analysis of random amplified polymorphic DNA (RAPD) showed that 43 S. integrifolium-specific and 57 S. sanitwongsei-specific bands were all found in 24 plants. Both somatic hybrids and its S1 plants were found to be resistant to bacterial wilt, and eggplant grafted these plants using for rootstocks were more productive than grafted mother plants. Now, S1 progenies are used for commercial eggplant production in Osaka Prefecture.

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Evaluation of the Antioxidant Potential and ldentification of Active Principles of Solanum nigrum L. on Antioxidant Defense Systems (까마중내 (Solanum nigrum L.) 항산화방어계의 항산화력 및 물질의 동정)

  • 임종국;정규영;정형진
    • Journal of Life Science
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    • v.11 no.6
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    • pp.509-516
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    • 2001
  • Enzymes and non-enzymatic antioxidants are involved in defense of oxgen free radical intermediates in all aerobic cells. The non -enzymatic antioxidants and antioxidant enzyme from the extracts of Solanum nigrum L. known to be anticancer medicinal plant were examined in other to utilize the discovery in natural products as cancer chem-opereventive agents. The DPPH(1,1-diphenyl-2-picryl-hydrazyl) free radical scavening activity on plant position of Solanum nigrum L. was the highest in root, with stem, whole plant, seed, leaf and flower, at higher activities respectively. In extraction methods, the DPPH free radical scavenging activity by circulating extraction with 80 % MeOH. The DPPH activity of L6 fraction by LH-20 column chromatography showed about 6.7 times higher than that of ethyl acetate-fraction. These were identified as phenolic compounds such as 2-6-methano-3-benzazocin-11-ol, 2[1H]-phyidinethione and 2-hydroxy -5-methyl-benzaldehyde. Peroxidase(POD) and superoxide dismutase(SOD) activities of stem and root were higher than that of other plant positions and those of plant positions according to growing stage were the highest in 60 days after seeding. The numbers of isozyme pattern of POD and SOD showed 10 hands and 5 bands, respectively, especially, 8 bands of POD and 3 bands of SOC showed a difference according to plant positions.

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Protoplast Culture by Electrofusion of Protoplasts from Solanum sisymbriifolium and Other Solanum Species

  • Kim Hag-Hyun;Jung Hye-Jin;Shin Un-Dong
    • Korean Journal of Plant Resources
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    • v.19 no.3
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    • pp.422-426
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    • 2006
  • This research was conducted to get the basic materials necessary to obtain the somatic hybrid plant between Solanum sisymbriifolium and other Solanum species (S. integrifolium and S. toxicarium). Regarding the formation of colony from the protoplast in S. sisymbriifolium, S. integrifolium and the fused protoplast mixture; for the S. sisymbriifolium, a colony was observed in F medium(Kao medium containing $5.0mg{\cdot}L^{-1}\;NAA,\;1.0mg{\cdot}L^{-1}$ 2,4-D and $1.0mg{\cdot}L^{-1}$ BA); and for the S. integrifolium, in G medium (a half strength MS medium containing 0.03 M sucrose, 0.4 M mannitol, $1.0mg{\cdot}L^{-1}\;NAA,\;1.0mg{\cdot}L^{-1}$ kinetin) respectively. In mixed cultured protoplast after electriofusion treatment, the cell division and colony formation were observed in both media F and G. For the shoot and root formation rate, there was no difference between the parent of each breed and mixed protoplast regardless of the medium. In the fused protoplast mixture of S. sisymbriifolium and S. toxicarium, a colony formation was also observed in both media F and H(a half strength MS medium containing 0.03 M sucrose, 0.4 M mannitol, $1.0mg{\cdot}L^{-1}\;NAA,\;1.0mg{\cdot}L^{-1}$ kinetin); and there was no difference in the shoot and root formation rate between the parent and the mixed protoplast.

Activity of Alkaline Phosphatase from the Mosquito, Culex pipiens pallens (홍모기(Culex pipiens pallens)의 난성숙 과정 중 alkaline phosphase의 활성)

  • 이영수;이승훈;박영민;성기창
    • The Korean Journal of Zoology
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    • v.36 no.3
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    • pp.425-432
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    • 1993
  • Alkaline phosphatase from Culex pipiens pallens was examined to determine the optimal assay condition and to assay the activity during ovarian development. The activity of alkaline phosphatase in a male and a nongravid female continuously were declined after eclosion. But by the stimulus of a blood meal, the enzyme activity was increased dramatically. At 30 hr. after a blood meal, the maximal activity was reached and then declined. And after 48 hr. after a blood meal, the second activity increase was revealed. This second increase was maintained up to oviposition. The first activity increase was revealed in the midgut and the second increase was done in the ovary to assay the organ distribution of alkaline phosphatase. In electrophresis data, it was shown 5 isozyme bands, ALP-1 and ALP-2 in the ovary, ALP-3 in the thorax and the midgut, and ALP-4 and ALP-5 in the thorax, the fatbody and the midgut in crude extract at 30 hr. after a blood meal. One the same ovary pattern were shown at 72 hr. after a blood meal.

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