• Title/Summary/Keyword: peroxidase isozyme

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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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Effects of Ethephon and Gibberellin on Sex Expression, and Subsequent Changes in Protein Contents, Peroxidase Activities, and Isoperoxidase Isoperoxidase Patterns of Cucumis sativus L. (Ethephon과 Gibberellin 처리(處理)가 오이의 성발현(性發現)과 이에 따른 단백질함량(蛋白質含量), Peroxidase 활성(活性) 및 Isoperoxidase Pattern에 미치는 영향(影響))

  • Ku, Woo Seo;Kim, Young Rae
    • Korean Journal of Agricultural Science
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    • v.12 no.1
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    • pp.1-16
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    • 1985
  • This experiment was conducted to study sex expression and subsequent changes in protein and peroxidase after three cultivars of cucumber plants were treated with ethephon and gibberellin. The three cultivars of cucumber used in this study included 'Sayeup' (monoecious type), 'Sinrokdadaki' (gynoecious type), and 'Seonghowon' (intermediate type). The ethephon at 250 ppm and gibberellin at 100 ppm were treated at the 2-leaf and 4-leaf stages, and subsequent sex expression and changes in protein contents, peroxidase activities and isoperoxidase banding patterns by disc polyacrylamide gel electrophoresis were studied. The results can be summarized as follows: 1. Ethephon treatment slightly increased number of pistillate flowers and significantly decreased number of staminate flowers in the three cultivars, while gibberellin treatment significantly increased number of staminate flowers in both gynoecious 'Sinrokdadaki' and intermediate 'Seonghowon' and did not increase number of staminate flowers in monecious 'Sayeup' 2. There were some differences among three cultivars in protein contents, protein banding and isoperoxidase banding patterns of seeds and germinating seeds. However, it was not obvious to differentiate monoecious from gynoecious cultlvars by these characters. 3. Protein contents in the leaves. and stem apex after ethephon and gibberellin treatment increased gradually at the 2-leaf stage, but decreased at the 4-leaf stage. Protein contents in stem apex at the 4-leaf stage without treatment were much higher in 'Sinrokdadaki' and 'Seonghowon' than in monoecious 'Sayeup'. Protein contents in the stern apex at the 4-leaf stage were increased in the ethephon-treated monoecious 'Sayeup' and decreased in 'Sinrokdadaki' and 'Seonghowon' compared with untreated plants. 4. Peroxidase activities in the leaves and stem apex gradually decreased at the 2- leaf stage, but increased at the 4-leaf stage. Peroxidase activities in stern apex at the 4-leaf stage were significantly increased by ethephon treatment. 5. The number of protein bands in the three cuitivars after treatment gradually decreased in leaves and stem apex at the 2-leaf stage, but increased in the 4-leaf stage. The protein banding pat terns of stern apex of the ethephon-treated monoecious 'Sayeup' at the 4-leaf stage were gradually shifted to the banding.

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High Light-Induced Changes in the Activities of Antioxidant Enzymes and the Accumulation of Astaxanthin in the Green Alga Haematococcus pluvialis

  • Park, Seul-Ki;Jin, Eon-Seon;Lee, Choul-Gyun;Lee, Mi-Young
    • Molecular & Cellular Toxicology
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    • v.4 no.4
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    • pp.300-306
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    • 2008
  • We investigated high light-induced alterations in antioxidant enzymes by exposing green vegetative cells of the alga Haematococcus pluvialis to excess irradiance to induce the production of astaxanthin, a carotenoid pigment. Total activity of catalase decreased approximately 70% after high light exposure, whereas glutathione peroxidase (GPX) activity was slightly enhanced. Total activity of superoxide dismutase and ascorbate peroxidase (APX) also slightly decreased. Overall, we did not observe dramatically elevated levels of antioxidant isozymes, although APXn, GPX2, and GPX3 isozyme increased slightly. ${H_2}{O_2}$ content increased about sixfold after high light exposure, demonstrating severe cellular oxidative stress, whereas lipid peroxidation was notably reduced. Concomitantly, astaxanthin accumulation increased about sevenfold. This result suggests that probably massively accumulated astaxanthin may be one of the antioxidant protector against high light stress.

Activities and Isozyme Profiles of Antioxidant Enzymes in Intercellular Compartment of Overwintering Barley Leaves

  • Baek, So-Hyeon;Kwon, In-Sook;Park, Tae-Il;Yun, Song-Joong;Kim, Jin-Key;Choi, Kyeong-Gu
    • BMB Reports
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    • v.33 no.5
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    • pp.385-390
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    • 2000
  • Overwintering plants should survive the various biotic and abiotic stresses that occur during winter. Previous studies indicated that active oxygen species are involved in freezing, dehydration, anoxia and pathogen infections. As the importance of the events that occur in the intercellular compartment became apparent in disease resistance, we examined the nature of intercellular antioxidant enzymes in order to access their possible involvement in the winter hardiness of barley. The levels of intercellular peroxidase, catalase, and SOD activities on the unit protein basis were 394, 18, and 9% of those of cellular activities, respectively. Major intercellular peroxidase isoforms consisted of four neutrals and four basic forms; whereas major cellular isoforms were two basic forms. Out of the two major catalase isoforms a higher molecular weight form was predominantly abundant in both cellular and intercellular compartments. Among the five major cellular SOD isoforms, three were also present in the intercellular compartment. The presence of substantial amounts of intercellular antioxidant enzymes in overwintering barley leaves may suggest the involvement of these enzymes in the tolerance mechanism to the various stresses that occur during winter.

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Varietal Classification of Barley by Isozymes and Restriction Fragment Length Polymorphisms (RFLPs) (동위효소 분석과 제한효소 단편 다형화현상을 이용한 보리 품종의 분류)

  • Jin, Byung-Soon;Park, Ro-Dong;Eun, Moo-Young;Lee, Eun-Seop
    • Applied Biological Chemistry
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    • v.36 no.3
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    • pp.139-145
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    • 1993
  • The methods of isoelectric focusing of 4 isozymes in polyacrylamide horizontal slab gels and restriction fragment length polymorphisms (RFLPs) were applied to characterize the biochemical phenotypes of 19 cultivars of barley. Among 19 barley cultivars screened, 7 esterase, 3 phosphoglucose isomerase, 4 peroxidase and 2 alcohol dehydrogenase isozyme phenotypes were distinguished by isoelectric focusing. When purified DNA of each cultivar was digested with restriction enzyme EcoRV and analyzed its RFLPs with barley DNA markers pMSU 51 or pMSU 71, two distinct RFLP patterns were shown. Based on the four isozymes and two RELP polymorphisms, 19 cultivars of barley were classified into 13 biochemical phenotypes. Phylogenetical relationships among 13 biochemical phenotypes classified were determined using Nei's F-statistics and the phylogenetic dendrogram was constructed.

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Effects of Various Ions on the Cellular and Secretory Isoperoxidases in Rice Suspension Culture

  • Lee, Mi-Young
    • BMB Reports
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    • v.30 no.6
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    • pp.379-384
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    • 1997
  • The effects of several ions on the specific activity and isozyme patterns of cellular and secretory isoperoxidases were studied in suspension-cultured cells of rice (Oryza sativa L.). Peroxidase release into the culture medium occurred in the absence of added calcium. The addition of calcium ion greatly stimulated the secretion of cationic isoperoxidases such as C2 and C3 into the medium: a maximum 11 fold increase of secretions occurred in the presence of 5 mM $CaCl_2$, and the secretion was accomplished within 1 hour after the addition of $CaCl_2$. About a 10 fold increase of the peroxidase secretion into the medium did occur with 0. 5% NaCl, whereas cellular isoperoxidase levels were reduced notably. About a 6 fold increase of the specific activity of cellular isoperoxidase was found in 5 mM $NiCl_2$-treated cell, while $NiCl_2$ had no effect on the secretion of peroxidase into the medium. Various concentrations of KCl did not change peroxidase secretion, but 5 mM $ZnCl_2$ reduced peroxidase secretion greatly. The major secretory isoperoxidases stimulated by $CaCl_2$, NaCl and cellulase were composed of cationic isoperoxidases C2 and C3, which were found to be localized in the cell wall of rice by examination of the enzyme in the protoplast. Furthermore, the secretion rates of secretory isoperoxidases were increased rapidly when cellulase was treated in the absence of the osmotic stabilizer of 0.4 M mannitol. These results suggest that the stimulations of secretory isoperoxidase levels seem to be due to the stimulation of secretion into the culture medium of rice.

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Effects of polyamines on hydrogen peroxide-scavenging enzymes in radish seedling plants under paraquat stress (Paraquat 스트레스를 받는 무 (Raphanus sativus L) 유식물에서 H2O2 분해 효소에 대한 폴리아민의 효과)

  • Jin, Chang-Duck
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.149-156
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    • 2009
  • Application of exogenous polyamines (PAs) reduced the paraquat (PQ)-induced cotyledon injuries in radish seedling plants with 1 mM spermidine (Spd) being the most effective protectant. PQ injury symptoms in the cotyledons, e.g., large accumulation of $H_2O_2$, and losses of fresh weight, chlorophyll, and proteins, were significantly alleviated. Likewise, analysis of $H_2O_2$-scavenging enzymes such as catalase (CAT) and guaiacol peroxidase (GPX) showed that pretreatment with Spd among PAs remarkably increased total CAT activity and strongly retarded PQ-induced rapid decline in total GPX activity. In a native gel assay, one CAT isozyme (CAT1) and two GPX isozymes (GPX1 and a newly synthesized GPX isozyme) proved to be more responsible for PQ tolerance, as manifested by the strong increases in their activities by Spd pretreatment. Based on these results, we can suggest that PAs (especially 1 mM Spd) may function as antioxidant protectors by invoking CAT and GPX enzymes which control the endogenous $H_2O_2$ level in radish cotyledons exposed to PQ.

Production of Antibacterial Substance, and Changes in Peroxidase nd Polyphenoloxidase Activities in Tobacco Plants Inoculated with Avirulent Isolate of Pseudomonas solanacearum (비병원성 Pseudomonas solanacearum 접종에 의한 담배내 항균물질생성과 Peroxidase 및 Polyphenoloxidase의 변화)

  • Yi Young Keun;Min Tae Gi;Park Won Mok
    • Korean Journal Plant Pathology
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    • v.3 no.3
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    • pp.203-209
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    • 1987
  • The substances obtained from the leaf, stem and root of tobacco plants inoculated with avirulent and virulent isolates of Pseudomonas solanacearum were at R_f\;0.6$ and R_f\;0.9$ on TLC plate, respectively. Both substances showed antibacterial activities not only on P. solanacearum but also on Erwinia carotovora subsp. carotovora and Escherichia coli in vitro. However, the antibacterial substances were not detectable from the filtrate of the autoclaves tobacco sap medium, in which the avirulent or virulent bacterium was cultured for 3 days. Peroxidase and poly phenoloxidase activities and their isozyme patterns did not differ significantly between plants treated with the virulent and avirulent isolates, or between the susceptible cultivar BY 4 and the resistant cultivar NC 82. However, activities of the two enzymes were increased in leaves of the susceptible cultivar BY 4 treated with either the virulent or the avirulent isolate.

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Transfer of Isolated Nuclei from Agrocybe aegerita Mycelia into Pleurotus florida Protoplasts (사철느타리버섯 원형질체내(原形質體內) 버들볏짚버섯 핵(核)의 전이(轉移))

  • Yoo, Young-Bok;Shin, Pyung-Gyun;Kim, Han-Kyoung;Byun, Myung-Ok;You, Chang-Hyun;Cha, Dong-Yeul;Chang, Kwon-Yawl
    • The Korean Journal of Mycology
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    • v.17 no.3
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    • pp.114-118
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    • 1989
  • The transfer of isolated nuclei from Agrocybe aegerita mycelia of wild type into Pleurotus florida protoplasts of auxotroph was induced with polyethylene glycol. The type 1 of nuclear transfer products was spontaneous segregants of both parental morphology of colony. Hyphae of A. aegerita type had clamp connections while that of P. florida type lacked. Type 2 was main products of nuclear transfer which formed true clamp connections. Type 3 was clampless products. They all produced primordia and developed basidiocarps similar to Agrocybe aegerita. A comparison of nuclei transfer products was made using isozyme analyses of alcohol dehydrogenase, esterase, lactate dehydrogenase and peroxidase. Isozyme band patterns of some type 2 strains produced a new band. Band patterns from mycelial extracts of the other strains could be characterized by parental bands.

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Studies on the Effect of Stress-Induced Gastric Ulcer on Gastric Carcinogenesis in Rats treated with N-methyl-N'-nitro-N-nitrosoguanidine (랫드에서 스트레스에 의한 위궤양이 위발암화과정에 미치는 영향에 관한 연구)

  • 이종권;김형진;이영순
    • Journal of Food Hygiene and Safety
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    • v.6 no.1
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    • pp.27-40
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    • 1991
  • ;ABSTRACT-The effects of gastric ulcer induced by restraint and water-immersion stress on gastric carcinogenesis in Wistar male rats treated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) were examined. Rats of group 1 were 2iven stress for 8 hours before they were received MNNG (100 mg/l) in drinking water for 20 weeks. Rats of group 2 were received MNNG first for 2 weeks and then were given stress once a week from 3rd to 12th weeks, with simultaneous MNNG adminitration and followed by MNNG only until 20th weeks. Rats of group 3 were received MNNG only as a positive control and rats of group 4 were not treated with carcinogen. All groups were sacrificed in 20 weeks. Sections of the pyloric mucosa were stained by avidin-biotin peroxidase complex (ABC) immuno-histochemical method. PAPG (pepsinogen isozyme 1 altered pyloric gland), body weight change, gross lesions and histopathological changes were examined. The results obtained from these studies were summarized as follows: 1. The mean body weight gains of the rats fed with carcinogens (group 1, 2, 3) were significantly lower than that of group 4 (control group, without carcinogen. p<0.05). However, the differences of the mean body weight of rats treated with carcinogen were not significant. 2. Stress treatment (group 1 and 2) increased the appearance of the numbers of PAPG (Pepsinogen 1 Altered Pyloric Gland) induced by carcinogen significantly compared with that of group 3 (carcinogen only, p<0.01). 3. The incidence rate of mucosal hyperplasia in pylorus was significantly increased in group 2 compared with group 3 (p<0.05).0.05).

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