• International Journal of Precision Engineering and Manufacturing-Green Technology
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• v.5 no.5
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• pp.631-636
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• 2018

Extraction of photosynthetic currents from thylakoids was studied using micro pillar structured electrode. Thylakoids were isolated from spinach leaves, and the size and shape of thylakoids were estimated from scanning electron microscopy images. Based on the geometry information of thylakoids, micro pillar shaped electrode was designed and fabricated using metal-assisted chemical etching of silicon wafers. Influence of photovoltaic effect on the silicon-based micro pillar electrode was confirmed to be negligible. Photosynthetic currents were measured in a three-electrode setup with an electron mediator, potassium ferricyanide. Photosynthetic currents from micro pillar electrodes were enhanced compared with the currents from flat electrodes. This indicates the significance of the enhanced contact between thylakoids and an electrode for harvesting photosynthetic electrons.

• Journal of Plant Biology
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• v.36 no.1
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• pp.97-104
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• 1993

The lipid composition of thylakoid membranes was compared between mesophyll and bundle sheath chloroplasts of maize. According to mild-denaturing gel electrophoresis, mesophyll thylakoids contained both PS I complex and PS II light-harvesting chlorophyll-protein complex(LHCP), while those of bundle sheath cells contained mainly PS I complex. The amount of lipids per mg chlorophyll was higher in bundle sheath thylakoids than in mesophyll. The major polar lipid classes were monogalactosyldiacylglycerol(MGDG), digalactosyldiacylglycreol, sulfolipid and phosphatidylglycerol (PG) in both tissues. Linolenic acid(18 : 3), linoleic acid(18 : 2) and palmitic acid(16 : 0) were the main fatty acyl components, with higher ratio of unsaturated to saturated fatty acids in bundle sheath thylakoids, suggesting these membranes are more fluid. The most striking difference in lipid composition between the two kinds of tissues was the practical absence of trans- 3-hexadecenoic acid(16 : 1t) in PG of bundle sheath thylakoids. This fatty acid is known to be involved in the association of LHCP as oligomeric form. More than 80% of MGDG molecular species was 18 : 3, 18 : 3, demonstrating that maize is a typical 18 : 3 plant. Therefore, the possibility of the functional relationships between the lamella structure, and thus the distribution of photosystems, and MGDG molecular species was excluded.

• Journal of Plant Biology
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• v.31 no.3
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• pp.217-226
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• 1988

Effects of simazine [2-chloro-4,6-bis(methylamino)-s-triazine] on the photochemical reactions of isolaed spinach chloroplasts and crude thylakoids of Anabaena inequalis UTEX B-381 were compared. Simazine inhibited photosynthetic O2 evolution and increased the chlorophyll fluorescence in whole cells of Anabaena. The electron transfer from diphenylcarbazide to 2,6-dichlorophenolindophenol was inhibited by simazine treatment in spinach chloroplasts, but not in crude thylakoids of Anabaena. In spinach chloroplasts, the chlorophyll fluorescence was increased by simazine treatment in the presence of diphenylcarbazide and ferricyanide, but not in the presence of diphenylcarbazide and silicomolybdate. In crude thylakoids of Anabaena, simazine treatment did not increase the chlorophyll fluorescence in the presence of either diphenylcarbazide and silicomolybdate, or diphenylcarbazide and ferricyanide. There results suggest that the inhibitory site of simazine on photosynthetic electron transport chain of anabaena is different from that of spinach chloroplasts. And there may be a possiblity that the inhibition site of simazine in Anabaena lies on the donor side of photosystem II, before the site of electron donation by diphenylcarbazide.

• Journal of Photoscience
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• v.9 no.2
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• pp.55-58
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• 2002

The reaction center Dl protein of photosystem II is the target of photodamage by excess illumination. The Dl protein is damaged by reactive oxygen species generated by photochemical reactions and then degraded by specific proteolytic enzymes. We found that the Dl protein also cross-links with the surrounding polypeptides, such as D2 and CP43 in isolated thylakoids or photosystem II-enriched membranes from spinach under the illumination with strong visible light. The cross-linking was observed in spinach leaf discs as well when they were illuminated at higher temperature (40°C). It was also shown that the cross-linked products are digested efficiently by a protease(s) in the stroma. Thus the cross-linking/digestion processes of the Dl protein seem to comprise a new pathway in the turnover of the photodamaged Dl protein. It should be noted, however, that the cross-linked products of the Dl protein and CP43 induced by endogenous cationic radicals in the donor-side photoinhibition are resistant to proteolytic digestion. Accumulation of these cross-linked products in the thylakoids may lead to the decay of the function of chloroplasts and finally to the death of plant cells. Thus, we suggest that the quality control of photosystem II, especially removal of the cross-linked products of the Dl protein, is crucial for the survival of chloroplasts under the light stress.

• 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.

• BMB Reports
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• v.29 no.1
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• pp.58-62
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• 1996

The structural changes in the electron donor side of the PSII reaction center have been monitored since heat treatment ($45^{\circ}C$ for 5 min) of thylakoids is known to decrease the oxygen evolving activity. In heat-treated spinach chloroplast thylakoids, the inhibitory effect of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) on the electron transport activity of the PSII reaction center from diphenyl carbazide to dichlorophenolindophenol became reduced approximately 3.8 times and [$^{14}C$]-labeled DCMU binding on the D1 polypeptide decreased to 25~30% that of intact thylakoid membranes, implying that the conformational changes of the DCMU binding pocket, residing on the D1 polypeptide, occur by heat treatment. The accessibility of trypsin to the $NH_2$-terminus of the cytochrome b-559 ${\alpha}$-subunit, assayed with Western blot using an antibody generated against the synthetic peptide (Arg-68 to Arg-80) of the COOH-terminal domain, was also increased, indicating that heat-treatment caused changes in the structural environments near the stromal side of the cytochrome b-559 ${\alpha}$-subunit, allowing trypsin more easily to cleave the $NH_2$-terminal domain. Therefore, the structural changes in the electron donor side of the PSII reaction center complexes could be one of the reasons why the oxygen evolving activity of the heat-treated thylakoid membranes decreased.

• ALGAE
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• v.18 no.2
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• pp.121-127
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• 2003

The pyrenoid ultrastructure and distribution of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and Rubisco activase in the chloroplasts of Chlamydomonas reinhardtii was studied using the immunogold localization technology with electron microscopy. There were several tubular thylakoids invading in the pyrenoid matrix to form several spokewise channels. The connections between pyrenoid matrix and stroma of chloroplast were the partial of channels. The starch sheath surrounding the pyrenoid was separated into several parts by the connections in transection. Some thylakoids were packed together near the connections in one side of the pyrenoid. Those special structures might be used to transport substance between pyrenoid and stroma of chloroplasts. With the antibody raised against the large subunits of Rubsico from C. protothecoides, the result of the gold immunolocalization of Rubisco in Chlamydomonas reinhardtii showed most of the gold particles heavily labeled the pyrenoid matrix, as well as the starch sheath matrix, and very few in the stroma of chloroplasts. The gold particle density was 880.00 $\pm$ 164.32, 190.00 $\pm$ 152.39 and 9.60 $\pm$ 5.37 ${\mu}m^{-2}$ in pyrenoid matrix, starch sheath and stroma region of chloroplast respectively (background: 5.67 $\pm$ 1.53 ${\mu}m^{-2}$). 99.59% of the total Rubiscos was calculated to be concentrated in the pyrenoid matrix and starch sheath by spatial densities. The gold immunolocalization of Rubisco activase also showed that Rubisco activase was mainly concentrated in the periphery of the pyrenoid and the starch sheath (the density was as high as 229.69 $\pm$ 96.96 ${\mu}m^{-2}$). There were very few gold particles located in the stroma of chloroplasts. These results indicated that pyrenoid surface and starch sheath was the site for Rubisco activation and $CO_2$ fixation, which supported the suggestion that pyrenoids perform photosynthesis function.

• Journal of Environmental Science International
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• v.15 no.12
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• pp.1093-1101
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• 2006

In the present study we studied the growth, photosynthetic traits and protective mechanisms against oxidative stress in the primary loaves of cucumber (Cucumis sativus L.) seedlings with or without UV-B treatment. Cucumber seedings were irradiated with UV-B for 10 days in environment-controlled growth chambers. The primary leaves irradiated with UV-B showed reduction in leaf length and decreased biomass production. The reduced biomass production seemed to be due to a negative effect of UV-B radiation on the photosynthetic process. Changes in chemical properties of leaf, such as chi a/b ratio affected photosynthesis. UV-B significantly affected chl b content compared with chi a in the light harvesting complex resulting reduced photosynthetic activity Fv/Fm decreased with an UV-B stress, suggesting that the photosynthetic apparatus, and particularly, PS II was damaged under UV-B stress. Malondialdehyde(MDA) concentration which represents the state of membrane lipid peroxidation Increased significantly under UV-B stress confirming an oxidative stress. UV-B exposure with SA solution(0.1-1.0 mM) can partially ameliorated some of the detrimental effects of UV-B stress. Leaf injuries including loss of chlorophyll and decreased ratio of Fv/Fm were reduced with combined application of UV-B and SA. ABA and JA showed similar mode of action in physiological effects on photosynthetic activities though the levels were lower than those from SA treated plants. Chloroplast ultrastructure was also affected by UV-B exposure. The thickness of leaf tissue components decreased and the number of grana and thylakoids was reduced in chloroplast applied UV-B or SA alone. At combined stress granal and stromal thylakoids were less affected. The leaves under combined stress acquired a significant tolerance to oxidative stress. From these results, it can be suggested that SA may have involved a protective role against UV-B induced oxidative damage.

• 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.

• Journal of Ginseng Research
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• v.14 no.2
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• pp.135-141
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• 1990

In order to elucidate the mechanism of the leaf-burning disease of ginseng (Panax ginseng C.A. Meyer), the relationships between thylakoid membrane peroxidation and chlorophyll bleaching were investigated in comparison with the ones of soybean (Glycine max L). When I measured the rate of lipid peroxidation in the thylakoids of ginseng and soybean by irradiation of light(60 w.m-2), it was identified that, the remarkably lower rate of lipid peroxidation was found in the ginseng thylakoid than the case of soybean. When lipid peroxidation of ginseng thylakoid was induced in the dark, chlorophyll contents of thylakoid was not changed. The results suggest that lipid peroxidation does not affect the chlorophyll bleaching in ginseng thylakoid. Thylakoid membrane peroxidation as well as chlorophyll bleaching was closely related with photosynthetic electron transport. But, according to the quenching experiment active oxygen species induced lipid peroxidation may be different species in the case of chlorophyll bleaching.