• Title/Summary/Keyword: thylakoid

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Changes in Protein Synthesis Induced by Chilling in Tomato Chloroplasts

  • Kim, Won-Il;Jung, Goo-Bok;Kim, Min-Kyeong;Park, Kwang-Lai;Yun, Sun-Gang
    • Korean Journal of Environmental Agriculture
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    • v.20 no.5
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    • pp.310-316
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    • 2001
  • To find out the effect of low temperature on the regulation of tomato chloroplast genes, the optimization of the system in chloroplast protein synthesis and the identification of the changes in chloroplast protein synthesis induced by chilling were studied. Incorporation reaction occurred rapidly at the first 30 minutes and was constantly maintained after 60 minutes. A broad optimal temperature on protein synthesis was found around 20 to $30^{\circ}C$. No difference was shown in the chloroplast protein synthesis under high light intensity (1600 ${\mu}E/m^2/s$) as well as under low light intensity (400 ${\mu}E/m^2/s$) even darkness. $K^+$, $Mg^{++}$ and ATP at an optimal concentration act as an activator, while DTT, chloramphenicol, cycloheximide, $Ca^{++}$ and inorganic phosphate act as an inhibitor in the chloroplast protein synthesis. Synthesis of 15, 55 and 60 kd chloroplast encoded stromal proteins and 18, 24, 33 and 55 kd chloroplast encoded thylakoid membrane proteins were reduced by chilling, while 17 kd chloroplast encoded stromal protein and 16 kd chloroplast encoded thylakoid membrane protein was induced by chilling. It was expected that the 55 kd stromal protein would be the large subunit of rubisco and the 33 kd thylakoid membrane protein would be the D1 protein which was drastically reduced by chilling.

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Glyphosate Toxicity: III. Detection of QB Protein in Thylakoid Membrane of Tomato Apical Meristem Using an Antibody Raised from Hybrid Protein of psb A and lac Z Gene (Glyphosate 독성: III. psb A와 lac Z 유전자의 Hybrid 단백질로부터 만들어진 항체를 이용한 토마토 정단분열조직의 Thylakoid막 내 QB 단백질의 검정)

  • Kim, Tae-Wan;Amrhein, Nikolaus
    • Korean Journal of Weed Science
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    • v.15 no.3
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    • pp.206-213
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    • 1995
  • Glyphosate(N-[phosphonomethyl]glycine) applied to the assimilate-exporting leaves(i.e. third old leaf) of tomato(Lycopersicon esculentum Mil var. Moneymaker). Herbicide binding protein, QB protein(D1), has been immunoblotted using the antibodies raised against the hybrid-protein expressed by a part of spinach psb A gene cloned in frame with the 3'end of lac Z gene to allow expression of the ${\beta}$-galactosidase(EC 3.21.23) in Escherichia coli. Glyphosate has an effect on a turnover of D1 within photosystem II of thylakoid membrane. The dysfunction of D1 protein within light harvesting complex(LHC-II) seems to be a pleiotropic effect of glyphosate.

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Effects of Light and Photosynthetic Electron Transport System on the Generation of Singlet Oxygen ($^1$O$_2$) in Ginseng Thylakoid Membrane (인삼 틸라코이드에서 Singlet Oxygen($^1$O$_2$) 생성에 미치는 전자전달계의 영향)

  • Yang, Deok-Cho;Chae, Quae;Lee, Sung-Jong;Kim, Yong-Hae;Kang, Young-Hee
    • Journal of Ginseng Research
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    • v.14 no.1
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    • pp.57-62
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    • 1990
  • In order to Investigate the mechanism of the leaf-burning disease of ginseng (Panax ginseng C.A. Meyer), studies on the generation of singlet oxygen (1O2) and the photooxidation of the pigments were carried out in comparison with the ones of soybean (G1ycine max L). The studies were mainly focalized on the effects of light intensity, light intensity, inhibitor and electron donor/acceptor of the Photosynthetic electron transport system. When we measured the amounts of 1O2 generated in the thylakoids of ginseng and soybean by the irradiation of light (300 w/m2) as a function its time. It was identified that a higher amount of 1O2 was formed in the ginseng thylakoid than the case of soybean. A generation ratio of lO2 between ginseng and soybean sltbstantially identical in the range of light intensities 50∼150w/m2 However much higher amount of 1O2 was generated in ginseng by irradiation of strong intensity of light (200 500w/m2). Wave length dependency on the generation of 1O2 and the pigment photooxidation was observed on ginseng thylakoids; red light (600-700 nm) gave a maximum effect in the contrast with blur green light (400-60 nm). When the ginseng thylalioid was treated with the electron donor (Mn2+) and acceptors (DCPIP, FeCy) of the photosynthetic electron transport system. a drastic inhibition of 1O2 generation was observed. However, treatment with its inhibitors (DCMU, KCW) activated 1O2 generation. An interesting fact that an electron donor or acceptor of the photosystem II(P680) Inhibited 1O2 generation, suggests an intimate relationship between 1O2 generation and photosystem II.

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Targeting Analysis of Lumenal Proteins of Chloroplast of Wheat using Proteomic Techniques

  • Kamal, Abu Hena Mostafa;Kim, Da-Eun;Oh, Myoung-Won;Chung, Keun-Yook;Cho, Yong-Gu;Kim, Hong-Sig;Song, Beom-Heon;Lee, Chul-Won;Uozumi, Nobuyuki;Choi, Jong-Soon;Cho, Kun;Woo, Sun-Hee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.05a
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    • pp.14-14
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    • 2010
  • Plastid proteomics are essential organelles present in virtually all cells in plants and green algae. Plastids are responsible for the synthesis and storage of key molecules required for the basic architecture and functions of plant cells. The proteome of plastid, and in particular of chloroplast, have received significant amounts of attention in recent years. Various fractionation and mass spectrometry (MS) techniques have been applied to catalogue the chloroplast proteome and its sub-organelles compartments. To better understanding the function of the lumenal sub-organelles within the thylakoid network, we have carried out a systematical analysis and identification of the lumenal proteins in the thylakoid of wheat by using Tricine-SDS-PAGE, and LTQ-ESI-FTICR mass spectrometry followed by SWISS-PROT database searching. We isolation and fractionation these membrane from fully developed wheat leaves using a combination of differential and gradient centrifugation couple to high speed ultra-centrifuge. After collecting all proteins to eliminate possible same proteins, we estimated that there are 407 different proteins including chloroplast, chloroplast stroma, lumenal, and thylakoid membrane proteins excluding 20 proteins, which were identified in nucleus, cytoplasm and mitochondria. A combination of these three programs (PSORT, TargetP, TMHMM, and TOPPRED) was found to provide a useful tool for evaluating chloroplast localization, transit peptide, transmembranes, and also could reveal possible alternative processing sites and dual targeting. Finally, we report also sub-cellular location specific protein interaction network using Cytoscape software, which provides further insight into the biochemical pathways of photosynthesis. The present work helps understanding photosynthesis process in wheat at the molecular level and provides a new overview of the biochemical machinery of the thylakoid in wheat.

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Stabilization of Membrane Proteins by Benzyladenine during Wheat Leaf Senescence (노쇠중인 밀잎에서 Benzyladenine에 의한 막단백질의 안정화)

  • 진창덕
    • Journal of Plant Biology
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    • v.35 no.2
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    • pp.117-123
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    • 1992
  • The effect of benzyladenine (BA) on lipid peroxidation and compositions of total insoluble proteins and chloroplast thylakoid protein from wheat primary leaves during senescence in the dark was studied. BA ($10^{-5}\;M$) treatment prevented conspicuously the loss of chlorophyll content and soluble and insoluble leaf protein contents in senescing wheat leaf segments during 4-day dark incubation. Under the BA treatment, especially, the level of insoluble protein was highly maintained than that of soluble protein. Also, the increase of malondialdehyde (MDA: the peroxidation product of membrane lipids) content was inhibited in the BA treated leaves. Three major polypeptide bands in quantity corresponding to 57, 26 and 12 KD molecular weight were clearly resolved with other minor bands by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) in the insoluble protein fraction. The insoluble protein profiles of the control leaves showed a remarkable decrease in the intensity of the 57 and 12 KD band except for 26 KD band in the 72 h dark incubation. This loss during dark incubation was reduced by BA treatment. More than 20 polypeptides were resolved in the chloroplast thylakoid membrane fraction with the most prominent bands which are 59 and 57 KD ($\alpha\;and\;\beta$ subunit of coupling factor: CF) and 26 KD (apoprotein of LHCP). The changes in thylakoid protein profile during 72 h dark incubation showed the rapid degradation in control, but this degradation was prevented in quantity by BA treatment. The above results suggested that BA would inhibit the peroxidation of membrane lipids, thereby preventing the loss of membrane proteins which led to the maintenance of the membrane integrity including chloroplast thylakoid.

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The Reason for the Loss of Photosynthetic Activity in Isolated Spinach Chloroplasts during Photosynthesis (분리된 시금치 엽록체에서 광합성중 광합성능이 소실되는 이유)

  • 김성수
    • Journal of Plant Biology
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    • v.36 no.4
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    • pp.371-375
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    • 1993
  • $ CO_2$ fixation of isolated intact spinach chloroplasts under saturating light began to decrease after 20 min, and stopped completely after 1 h. To identify the lesion sites for the die off, reconstituted chloroplast system was used with chloroplasts collected at several phases of time course. $ CO_2$ fixation was inhibited in the reconstituted chloroplasts made of thylakoids and stroma in the later phases, but showed a higher degree of inhibition by the participation of thylakoids than that of stroma in the later phases. Measurement of photophosphrylation and NADP reduction revealed that a severed thylakoidal damage was occurred at the later phases. This results indicate that the lesion sites for the die off are in the thylakoid.the thylakoid.

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Analysis of Chloroplast Mutants of Arabidopsis Induced by Diepoxybutane (Diepoxybutane에 의해 유도된 애기장대 엽록체 돌연변이체의 분석)

  • 윤용휘;이정훈;박해진;강용원;이경민;신동현;이인중;김학윤;김달웅
    • Journal of Life Science
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    • v.12 no.4
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    • pp.399-406
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    • 2002
  • The Arabidopsis mutants involved in chloroplast development were induced by seed treatment of diepoxybutane which was rarely known mutagenic compound in plant mutagenesis. Three kinds of mutants designated as iml, gev, and yev were represented by the characteristics of variegated leaves, green vein with yellow leaves, and yellow green vein with green leaves respectively. We investigated the ultrastructure of chloroplast in mutated regions using transmission electron microscopy. The ultrastructure of chroloplast in wildtype showed regularly stacked grana thylakoid and stroma thylakoid while iml, gev and yev mutants displayed different shapes of grana stacking and stroma stacking of chloroplasts. Genetic analysis of three chloroplast mutants exhibit that divergent traits were ruled by a single recessive nuclear gene.