• Title/Summary/Keyword: isoperoxidase $A_3$

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Effects of Salicylate on the Activity of Isoperoxidase $A_3$ from Tobacco Callus (Salicylate가 담배 미분화세포 isoperoxidase $A_3$의 활성에 미치는 영향)

  • 이미영
    • Korean Journal of Environmental Biology
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    • v.19 no.3
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    • pp.211-217
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    • 2001
  • Salicylate is involved in the induction of pathogen-related proteins and plant defense response. The effects of salicylate on the activity isoperoxidase $A_3$ from tobacco callus (Nicotiana tabacum L.) and the protection against the enzyme inactivation by salicylate in the presence of $Fe^{2+}$ were examined. About 20% and 85% activity losses of peroxidase occurred at 0.48 mM and 0.6 mM salicylate, respectively, showing that isoperoxidase $A_3$ was inactivated by salicylate. The inactivation occurred depending on pH and showed noncompetitive inhibition mode. Moreover, inactivation of the enzyme by salicylate was completely protected in the presence of $Fe^{2+}$. Apoperoxidase without heme moiety was constructed and the effects of various metal ions on the recovery of enzyme activities were investigated. More than 80% of the activity was reconstituted by the addition of $Fe^{2+}$ or hemin. However, the enzyme activity was not recovered by $Cu^{2+},\;Zn^{2+},\;Co^{2+},\;or\;Mn^{2+}$.

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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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Purification and Pysiological Characterization of Isoperoxidase from Oat Root Treated with Alachlor (Alachlor 처리후(處理後) 귀리 근단(根端)에 존재(存在)하는 동위과산화효소(同位過酸化酵素) 정제(精製) 및 효소(酵素)의 생리적(生理的) 특성(特性))

  • Kwon, S.W.;Han, K.S.;Kim, J.C.
    • Korean Journal of Weed Science
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    • v.14 no.1
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    • pp.56-61
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    • 1994
  • The cationic isoperoxidases were isolated from oat root tips which had been grown in treatment with $1{\times}10^{-4}$ M alachlor and purified about 30-fold by treatment with ethylalchol and ion exchange chromatography on DEAF-celluose and CM-sephadex medium. The oat root was found to contain three isoperoxidase. The major activity peak (B) represented 65% of the total isoperoxidase activity. After purification, the major peak of isoperoxidase was purified about 37-fold from the oat root. Analysis of the major peroxidase peak(column fraction 58-78) by SDS revealed a single band which corresponed to a molecular mass of 42.5 kD. In vitro, isoperoxidase activies were inhibited by IAA. Isoperoxidase(50 unit) significantly inhibited 70.2% of cell division in oat root and 54.2% of cell elongation in oat coleoptile as compared with control.

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Studies on Peroxidase Isozymes of Tobacco and Korean Radish: implication of Specific Isoperoxidase in Shoot Formation (담배와 무 Peroxidase Isozyme들의 세포 생화학적 연구: 조직 특이성 발현과의 연관성)

  • 김승수
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.357-389
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    • 1987
  • In an attempt to clarify the physiological functions of individual isoperoxidases, we have studied enzymatic and immunological properties as well as cellular distribution of isoperoxidases from tobacco callus and Korean radish. The gene expression patterns of isoperoxidases in shoot and non-shoot-forming tobbaco callus were also examined by rabbit reticulocyte lysatein vitro translation system. These results indicate that fraction of translatable poly(A)-isoperoxidase mRNA was increased considerably in shoots. At the present time, at least 6-7 isoperoxidases could be detected from the translation mixture of total cellular RNA, among which only one cell wall localized anodic isoperoxidase (named A3) mRNA was bimorphic mRNA. These data suggest the possible regulation of peroxidase activity during shoot formation by altering the polyadenylation state of mRNA. In case of Korean radish seedlings, poly(A)- peroxidase mRNA were also increased depending upon aging.

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Effect of $GA_3$ and ABA on Peroxidase and Catalase Activities and Isoperoxidase Patterns in Mung Bean Seedling (녹두의 발아과정 중 $GA_3$ 및 ABA의 처리가 Peroxidase, Catalase 활성변화와 Isoperoxidase Pattern에 미치는 영향)

  • Lee, Sang-Kap;Park, Woo-Churl
    • Applied Biological Chemistry
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    • v.31 no.2
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    • pp.205-210
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    • 1988
  • The changes in peroxidase and catalase activities, and isoperoxidase patterns in different parts of mung bean seedling caused by the treatment with plant growth substances, $GA_3$ and ABA, were examined. As germination proceeded, the activity of peroxidase in all part except hypocotyl was increased, while that of catalase decreased. The separate application of $GA_3$ and ABA increased the activity of peroxidase which was more influenced by $GA_3$ than by ABA only in cotyledon, while that of catalase was more affected by ABA than by $GA_3$. Electrophoretic study revealed that the number of isoperoxidase was increased continuously in all parts during development. A greater influence was exerted on the intensity of isozyme than the number of isozyme by the hormonal treatment.

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Characteristics of a Low Molecular Weight Minor Anionic Isoperoxidase $A_{3n}$ from Radish

  • Lee, Mi-Young;Kim, Soung-Soo
    • BMB Reports
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    • v.31 no.6
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    • pp.548-553
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    • 1998
  • A minor anionic isoperoxidase named $A_{3n}$, was isolated from Korean radish (Raphanus sativus L.) root. Purification of the enzyme was accomplished by CMcellulose chromatography, DEAE-Sephacel chromatography, and Sephadex G-75 gel filtration. The enzyme was a glycoprotein with molecular weight of approximately 31,000 as determined by SDS-PAGE and 33,000 by Sepadex G-150 gel filtration, which is by far the smallest among the reported isoperoxidases. The pI value was 3.5. The optimum pH of the enzyme was 6.5 for guaiacol and $H_2O_2$, and the $K_m$ values for guaiacol and $H_2O_2$ were 13.3 mM and 1.5 mM, respectively. Kinetic studies with various substrates revealed that only A3n, unlike other isoperoxidases from radish, did not use scopoletin as a substrate and had very low $K_m$ value of 0.25 mM for ferolic acid among naturally occurring phenolic substrates.

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The Changes of Peroxidase Activity and Isoperoxidase Patterns from Pine Needles under the Salinary Stress (염분스트레스에 의한 소나무잎 Peroxidase의 활성 및 Isozyme Pattern의 변화)

  • 이미영
    • The Korean Journal of Ecology
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    • v.20 no.5
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    • pp.315-321
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    • 1997
  • Peroxidase activities and isozyme patte군 of the pine needles (Pinus densiflora) were examined and compared in the coastal regions of Anmyum-Do(Choongnam, Taean-Gun) and inland regions of Shinchang-Myun(Choongnam, Asan-City). The pine needle peroxidase from Anmyum-Do showed approximately three times higher specfic activity than Shinchang pine needle peroxidase. The pine needle extracts of Anmyun-Do and Shinchang contained three anionic isoperoxidases, named A1, A2 and A3, when subjected to starch gel electrophoresis at pH 7.0. Cjationic isoperoxidases could not be found in both extracts., However, there existed unique isoperoxidase An only from the extracts of Anmyun-Do pine needles under the salinary environment. Moreover, the specific activities of catalase and glucose-6-phosphate dehydrogenase from Anmyun-Do, known for the inducible enzymes under the stress condition, were about 1.8 times higher than those of Shinchang pine needles. However, the specific activities of other enzymes did not show great differences between the two regions. Considering the above results of the higher specific activity of peroxidase and the unique expression of isoperoxidase An, pine needle peroxidase might involve in the defence mechanism against the salinary stress of Anmyun-Do.

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A Study of Mode of Action of Alachlor - II. Effect of Alachlor on Peroxidase Synthesis in Oat(Avena sativa L.) (Alachlor의 제초기구(除草機構)에 관한 연구(硏究) - II. Alachlor가 귀리의 Peroxidase합성(合成)에 미치는 영향(影響))

  • Kwon, S.W.;Kim, J.C.
    • Korean Journal of Weed Science
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    • v.10 no.3
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    • pp.233-239
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    • 1990
  • The effect of alachlor treatment on peroxidase synthesis in oat root tips was studied. Alachlor caused increase in the amount of soluble peroxidase in oat root tips, peroxidase activity increase as the rate of alachlor application increased, Alachlor treatment of oats with $1{\times}10^{-6}M$, peroxidase activity increased 0.20 unit higher than that of nontreatment. After 12hr, 65mM of peroxide treatment of oats inhibited 16% root growth, and 130 mM peroxide treatment caused 59% inhibition. With PAGE of peroxidase extracted from normal root tips, PAGE give 4 species($P_2$, $P_3$, $P_4$, and $P_5$ band) of peroxidase. Alachlor significantly activated isoperoxidase. Three isoperoxidase($P_1$, $P_6$, and $P_7$) are synthesized at a increased concentration of alachlor, SDS-PAGE analysis of proteins extracted from oat root tips showed that they were made up of subunits blow 100 kD polypeptide.

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Use of Zymography for Identification of the Same Clone in a Clone Bank of Pinus densiflora Sieb. et Zucc (Isoperoxidase 변이형(變異型)에 의(依)한 소나무 Clone 감별(鑑別))

  • Park, Young-Goo;Choi, Jung Suk
    • Journal of Korean Society of Forest Science
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    • v.18 no.1
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    • pp.17-22
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    • 1973
  • Using of zymography for identification of same clone of Pinus densiflora, the two year old needle leaves of 48 ramtes including 8 clones (6 ramets per clone) were collected in the clone bank which has been established on the near campus of Institute of Forest Genetics, Suwon, in 1962. All 8 bands are named from cathod to anode, G, H, K, M, N, Q, S and Y. Only CB-1 clone shows all bands, while KW-3 clone reveals only 5 bands. Other 6 clones were found 7 bands but the occurred frequencies of those bands are variable among those clones. Though the grafting stock are used various individuals grown seed propagation of P. densiflora, moreover, three stocks have been different species and that one has been P. rigida and two individuals have been P. koraiensis, the zymograms of the ramets belonging to the same clone reveals the identified patterns. The results show that the stocks for grafting have not been affected on the isoperoxidase patterns of their scions in P. densiflora. Among 48 ramets of 8 clones, 4 ramets are found the different isoperoxidase patterns from that of the remained rametes within same clone. Thus, it is conluded that zymography is useful for testing genuineness of the grafted clones of P. densiflora.

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Cytogenetic and Physiological Studies in Natural Populations of Torreya nucifera (비자나무 천연생집단(天然生集團)의 세포유전학적(細胞遺傳學的) 및 생리학적(生理學的) 연구(硏究))

  • Kim, Yeung Du;Kwon, Yeong Cheol
    • Journal of Korean Society of Forest Science
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    • v.78 no.1
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    • pp.42-54
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    • 1989
  • These studies were carried out to examine the anatomical, karyological characteristics, the variation of isozymes in needle and the frost damage of tissue for the purpose of investigating the relationships among the three natural Torreya nucifera populations of Cheju-do, Namhae, and Mt. Naejang in Korea. The results obtained can be summed up as follows : 1. The numbers of endodermal cells and inner layer cells of mucilage canal of needle conductive tissue were different in each population. The number of those of Cheju-do population was the largest. 2. In somatic chromosome, Namhae population showed one more secondary constriction. The values of $b^{arm}/a^{arm}$ were the same in all the three populations, but Cheju-do population particularly showed a different minimum value. And the karyotype formulas of each population showed difference. 3. During the meiosis, each population showed no significant difference in the irregular phenomena of chromosome. 4. In isoperoxidase and esterase variations of needle, each population showed its particular number and variation of bands. Cheju-do population showed the largest number and greatest variation of bands. 5. Under the same freeging conditions, the frost damage of Mt. Naejang population was comparatively slight, and Cheju-do population suffered from a greater frost damage than the others.

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