• Title/Summary/Keyword: isozyme profile

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Activity and Isozyme Profile of Antioxidative Enzymes at Booting Stage of Rice Treated with Cold Water

  • Kim Ki-Young;Kim Bo-Kyeong;Shin Mun-Sik;Choung Jin-Il;Ko Jae-Kweon;Kim Jung-Kon;Lim Jung-Hyun;Yun Song-Joon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.4
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    • pp.289-294
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    • 2004
  • This study was carried out to investigate the antioxidative enzymes and isozymes between chilling-tolerant and -susceptible varieties at the booting stage under cold water stress $(13^{\circ}C)$ in japonica rice. Total SOD, CAT, POX, and GR activities on the basis of protein were found to be important factors to defend cold water stress. Especially, SOD and CAT activities showed distinctive differences between chilling-tolerant and -susceptible varieties. Chilling-tolerant varieties were higher than chilling-susceptible varieties for SOD and CAT activities. One of eight isozyme bands for SOD was a inducible isoform. Three isozymes for CAT and one isozyme for POX were closely correlated with defense to cold water stress. Total GR activities except Stejaree 45 on the basis fresh weight and POX were increased by cold water stress, but there was no difference between chilling-tolerant and -sus­ceptible varieties.

Studies on Protein Profiles and Isozymes in Germinating Seeds (종자발아에 있어서 Protein Profile과 Isozyme에 관한 연구)

  • 권오용
    • Journal of Plant Biology
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    • v.17 no.4
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    • pp.143-156
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    • 1974
  • The purpose of this experiment was to study one side of germination physiology based on that protein profiles and protease relating to protein metabolism, that peroxidase, catalase, $\alpha$-amylase, $\beta$-amylase, and malate dehydrogenase involved in the carbohydrate metabolism of seed germination. All these experiments were divided into the two groups with and without acetone treatment, and were carried out. The protein bands of each germinating stage between the groups treated with and without acetone showed certain basic pattern in polyacrylamide gel disc electrophoresis. However, there was a little difference in the number of protein band, optical density, and migration velocity between two groups. The isozyme bands of peroxidase, and catalase between two groups in polyacrylamide gel disc electrophoresis did not show the numeral difference, but the optical density of certain germinating stage treated with acetone was higher than the group untreated with it and it showed their enzyme activity. The $\alpha$-amylase and $\beta$-amylase activities which involved in starch metabolism of seed germination were higher in the treated group than the other. On one hand, the protease activity of hydrolase occurred in the seeds for germination was also higher, more or less in the treated group than in the other. The isozyme band pattern of malate dehydrogenase in TCA cycle of energy metabolism pathway was very different between two groups growing for 72 hours with and without acetone treatment in cellulose acetate electrophoresis. It indicated that two isozyme bands of malate dehydrogenase was high. Consequently these experimental results mentioned above indicated that acetone treatment before sowing had an effect on dissolving certain complexed lipid substance involved in the seed coats, the activity of carbohydrate hydrolase increased with water absorption which was most comfortable in its germination, dissolved glycerin and fatty acid became certain energy source, and they stimulated the acceleration of respiration metabolism.

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Overproduction of Lignin Peroxidase from Phanerochaete chrysosporium PSBL-1 (Phanerochaete chrysosporium PSBL-1의 배양조건 최적화를 통한 Lignin Peroxidase의 과량생산)

  • 정병철;한윤전;장승욱;정욱진;원유정
    • Korean Journal of Microbiology
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    • v.36 no.3
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    • pp.228-235
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    • 2000
  • Until now, it was diIliculi to overproduce lignin peroxidase(LiP) fiom Pl~anemchaete ch~ysosporium since the lack of optimized growth conditions. In this paper, we optimized the LIP production conditions and monitored LIP isozyines of fl chqsospoi.ium PSBL-1. The optimized condition includes sponge matrix support, no addition of $MnSO_4$, excess addition of niixogen source(48 inM diarmnonium), and addition of stabilizer(2 mM verakyl alcohol). Finally we obtained Lip activity of 1,800 unitsll. HI isozyne was overproduced when inyceliuin was cultivated in media containing $Mn^{2+}$ (2.73 inM) and excess nitrogen(48 11d4 diannnonium). Three azo dyes(acid yellow 9, congo ued, orange IT; each concenimtion of50 $\mu$M) we1-e rapidly decolorized within 2 inins by 0.4 un~t or Lip.

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Mechanisms of Chilling Tolerance in Relation to Antioxidative Enzymes in Rice

  • Kuk, Yong-In;Shin, Ji-San;Whang, Tay-Eak;Guh, Ja-Ock
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47 no.5
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    • pp.341-351
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    • 2002
  • In order to examine the mechanistic basis for differential sensitivities to chilling and subsequent recovery between two rice (Oryza sativa L.) cutivars, a chilling-tolerant japonica type (Ilpumbyeo) and a chilling-susceptible indica type (Taebaekbyeo), changes of physiological responses and antioxidant enzymes were investigated. Both cultivars at 3 leaf stage were exposed at a low temperature of $5^{\circ}C$ for 3 days and subsequently recovered in a growth chamber at a $25^{\circ}C$ for 5 days with 250 mmol $m^{-2}$ $s^{-1}$. Physiological parameters such as leaf fresh weight, relative water content, cellular leakage, lipid peroxidation, and chlorophyll a fluorescence showed that the chilling tolerant cultivar had a high tolerance during chilling. However, the chilling-susceptible cultivar revealed severe chilling damages. The chilling-tolerant cultivar was also faster in recovery than the chilling-susceptible cultivar in all parameters examined. We analyzed the activity and isozyme profiles of four antioxidant enzymes which are: superoxide dismutase (SOD), caltalase (CAT), ascorbate peroxidase (APX), and glutation reductase (GR). We observed that chilling-tolerance was due to a result of the induced or higher antioxidant enzyme system, CAT and APX in leaves and SOD, CAT, APX, and GR in roots. Especially, we observed the most significant differences between the chilling-tolerant cultivar and -susceptible cultivar in CAT and APX activity. Also in isozyme profiles, CAT and APX band intensity in the chilling-tolerant cultivar was distinctively higher than in the chilling-susceptible cultivars during chilling and recovery. Thus, the cold stability of CAT and APX are expected to contribute to a tolerance mechanism of chilling in rice plants. In addition, the antioxidative enzymes activity in roots may be more important than in that of leaves to protect chilling damage on rice plants.

Generation of Nucler Hybrids Overcoming the Natural Barrier of Incompatibility: Transfer of Nuclei from Lentinula edodes into Protoplasts of Coriolus versicolor

  • Kim, Chaekyun;Choi, Eung-Chil;Kim, Byoung-Kak
    • Archives of Pharmacal Research
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    • v.23 no.1
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    • pp.79-86
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    • 2000
  • Heterokaryotic nuclear hybrids overcoming the natural barriers of incompatibility have been studied in basidiomycetes. To produce these nuclear hybrids between incompatible mushrooms, which have several potent pharmacological effects, nuclear transfer was performed between Lentinula edodes and Coriolus versicolor. Nuclei from serine auxotrophs of Lentinula edodes, $LE207(Ser^{-})$ were transferred into the protoplasts of arginine auxotrophs of Coriolus versicolor, $CV17(Ser^{-})$using 30% polyethylene glycol 4000 in 10 mM $Cacl_{2}$-glycine solution (pH 8.0). Nulcear transfer progenies were selected by nutritional complementation on minimal media supplemented with 0.6 M sucrose. The progenies were classified based on colony morphology to L. edodes-like, C, versicolor-like and non-parental type. Most of the progenies grew slower than either parent. The number of nuclei per cell was similar but the DNA content varied between progenies. The isozyme patterns of nuclear hybrids resembled either of the parent porfiles or showed a mixed profile.

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The RpoS Sigma Factor Negatively Regulates Production of IAA and Siderophore in a Biocontrol Rhizobacterium, Pseudomonas chlororaphis O6

  • Oh, Sang A;Kim, Ji Soo;Park, Ju Yeon;Han, Song Hee;Dimkpa, Christian;Anderson, Anne J.;Kim, Young Cheol
    • The Plant Pathology Journal
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    • v.29 no.3
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    • pp.323-329
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    • 2013
  • The stationary-phase sigma factor, RpoS, influences the expression of factors important in survival of Pseudomonas chlororaphis O6 in the rhizosphere. A partial proteomic profile of a rpoS mutant in P. chlororaphis O6 was conducted to identify proteins under RpoS regulation. Five of 14 differentially regulated proteins had unknown roles. Changes in levels of proteins in P. chlororaphis O6 rpoS mutant were associated with iron metabolism, and protection against oxidative stress. The P. chlororaphis O6 rpoS mutant showed increased production of a pyoverdine-like siderophore, indole acetic acid, and altered isozyme patterns for peroxidase, catalase and superoxide dismutase. Consequently, sensitivity to hydrogen peroxide exposure increased in the P. chlororaphis O6 rpoS mutant, compared with the wild type. Taken together, RpoS exerted regulatory control over factors important for the habitat of P. chlororaphis O6 in soil and on root surfaces. The properties of several of the proteins in the RpoS regulon are currently unknown.

Electrochemical properties of the mugwort-embedded biosensor for the determination of hydrogen peroxide (쑥을 이용한 과산화수소 정량 바이오센서의 전기화학적 성질)

  • Lee, Beom-Gyu;Park, Sung-Woo;Yoon, Kil-Joong
    • Analytical Science and Technology
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    • v.19 no.1
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    • pp.58-64
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    • 2006
  • A mugwort-tissue-based modified carbon paste electrode was constructed for the amperometric detection of hydrogen peroxide and its electrochemical properties are described. Especially the amperometric signal was very stable and bigger than any other enzyme electrode studied in this lab. The effect of tissue composition on the response was linear within the wide range of experiment and the linearity of Lineweaver-Burk plot showed that the sensing process of the biosensor is by enzymatic catalysis. And pH dependent current profile connoted that two isozymes are active in this system.

Genetic Diversity Studies and Identification of Molecular and Biochemical Markers Associated with Fusarium Wilt Resistance in Cultivated Faba Bean (Vicia faba)

  • Mahmoud, Amer F.;Abd El-Fatah, Bahaa E.S.
    • The Plant Pathology Journal
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    • v.36 no.1
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    • pp.11-28
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    • 2020
  • Faba bean (Vicia faba L.) is one of the most important legume crops in Egypt. However, production of faba bean is affected by several diseases including fungal diseases. Fusarium wilt incited by Fusarium oxysporum Schlecht. was shown to be the most common wilt disease of faba bean in Assiut Governorate. Evaluation of 16 faba bean genotypes for the resistance to Fusarium wilt was carried out under greenhouse conditions. Three molecular marker systems (inter-simple sequence repeat [ISSR], sequence related amplified polymorphism [SRAP], and simple sequence repeat [SSR]) and a biochemical marker (protein profiles) were used to study the genetic diversity and detect molecular and biochemical markers associated with Fusarium wilt resistance in the tested genotypes. The results showed that certain genotypes of faba bean were resistant to Fusarium wilt, while most of the genotypes were highly susceptible. The percentage of disease severity ranged from 32.83% in Assiut-215 to 64.17% in Misr-3. The genotypes Assiut-215, Roomy-3, Marut-2, and Giza2 were the most resistant, and the genotypes Misr-3, Misr-1, Assiut-143, Giza-40, and Roomy-80 performed as highly susceptible. The genotypes Assiut-215 and Roomy-3 were considered as promising sources of the resistance to Fusarium wilt. SRAP markers showed higher polymorphism (82.53%) compared with SSR (76.85%), ISSR markers (62.24%), and protein profile (31.82%). Specific molecular and biochemical markers associated with Fusarium wilt resistance were identified. The dendrogram based on combined data of molecular and biochemical markers grouped the 16 faba bean genotypes into three clusters. Cluster I included resistant genotypes, cluster II comprised all moderate genotypes and cluster III contained highly susceptible genotypes.

Difference in Electrophoretic Phenotypes of rice Cultivars Selected to Bensulfuron (Bensulfuron에 대(對)한 내성(耐性) 및 감수성(感受性) 수도품종(水稻品種)의 전기영동(電氣泳動) 표현형(表現型) 차이(差異))

  • Kuk, Y.I.;Guh, J.O.;Kim, Y.J.;Lee, D.J.
    • Korean Journal of Weed Science
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    • v.8 no.3
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    • pp.250-257
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    • 1988
  • The study was intended to know any relations between the rice tolerance to bensulfuron and varietal speciation in seed protein composition or any enzymatical allelies with or without chemical treatment. Rice varieties used were UCP-28, Chinsurah Boro II, Fukunohama, Fadehpur-2, IR 14252-13-2-2-5 as the tolerant group, and HP 93(3) FA, HP94(9) FA, Padilabou Alumbis, KH-17854, and IR 1846-2841-1 as the susceptible, respectively. Electrophoretic methods used were SDS-PAGE for seed protein, 7% PAGE for isozymes (acid phosphatase, peroxidase, malate dehydrogenase, and esterase from rice seedling) and variation in isoenzyme profiles (malate dehydrogenase, peroxidase, and esterase) as affected by different concentrations of bensulfuron(0, $10^{-6}$, $10^{-5}$ and $3{\times}10^{-5}M$) was also studied. The results are summarized as follows. -Among 16 bands separated in seed proteins, two different rice groups selected in terms of tolerance to bensulfuron were clustered in dissimilarity, which was based on relatively larger area in whole peaks and higher activities in N, O, P bands for the tolerant group. -Among isozymes obtained from rice seedlings without chemical treatments, the following specificities were obtained. The tolerant varieties had the relatively higher activity in D band out of 4 peroxidase bands. Malate dehydrogenase was separated into 3 bands and only tolerant varieties had A band and higher activities in Band C bands. Esterase was separated into 3-4 bands with higher activities in A and B bands for tolerant varieties. There were one major band accompanied by 2-3 minor bands for acid phosphatase in which only tolerant varieties had the B band. -The effect of Bensulfuron concentration on the isozyme activities showed that the activity of C band in peroxidase was not present in tolerant varieties which was contrary to the increased activities in susceptible varieties. However, D band was gradually disappeared only in susceptible varieties as the concentration of bensulfuron was increased. For malate dehydrogenase in the susceptible varieties, major bands D, E and F kept consistantly higher activities while minor bands A, B and C disappeared sensitively. Among 5 bands of esterase separated, D band was present only in the tolerant varieties while E band only in the susceptible. The activities in A, C, E bands were sharply decreased in the susceptible varieties as the concentration of bensulfuron was increased.

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Germination and Proteome Profile Characteristics of Wheat Seeds Treated under Different Concentrations of Abscisic Acid (Abscisic acid 농도에 따른 밀 종자의 발아와 단백질체의 발현 특성)

  • Jeong, Jae-Hyeok;Kim, Dae-Wook;Hwang, Woon-Ha;An, Sung-Hyun;Jeong, Han-Yong;Lee, Hyeon-Seok;Choi, In-Bea;Choi, Kyung-Jin;Yun, Jong-Tak;Yun, Song Joong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.63 no.1
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    • pp.25-34
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    • 2018
  • This study was conducted to investigate the germination and proteome profile characteristics of wheat seeds treated under various concentrations of abscisic acid (ABA). After-ripening, the seeds of three wheat cultivars (Baegjoong, Keumkang, and Uri) showing different levels of dormancy were used. Germination index and germination rate of the cultivars was higher than 0.95% and 98%, respectively, and these were not significantly different under 0, 10, 30, and $50{\mu}M$ ABA at 7 d after germination. However, the growth of the shoot and radicle was significantly inhibited at 10, 30, and $50{\mu}M$ ABA compared to that at $0{\mu}M$ ABA. Mean ABA content of the embryos of seeds germinated at 0 and $50{\mu}M$ ABA for 7 d was 0.8 and $269.0ngmg^{-1}DW$, respectively. Proteins extracted from embryos germinated for 4 d were analyzed by two-dimensional gel electrophoresis, and proteins showing a difference of 1.5-fold or greater in their spot volume relative to that of $0{\mu}M$ ABA were identified. The expression of four protein spots increased at $50{\mu}M$ ABA and two protein spots were detected only at $50{\mu}M$ ABA; these six proteins were all identified as globulin types. Conversely, the expression of three protein spots decreased at $50{\mu}M$ ABA and were identified as cytosolic glutamine sysnthetase, isocitrate dehydrogenase, and S-adenosylmethionine synthetase 2. In conclusion, ABA did not inhibit the germination rate regardless of pre-harvest sprouting characteristics of the cultivars. However, the growth of the shoot and radicle was significantly inhibited by ABA, most likely through the down regulation of glutamine, methyl group donor, and polyamines biosynthesis, among others, while accompanied by globulin accumulation in the embryos.