• 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.

• Journal of Plant Biology
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• v.37 no.2
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• pp.231-236
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• 1994

The effect of $Cd^{2+}$ on the electron transport rate of PSI and PSII was investigated in isolated spinach chloroplasts. In photosystem II, the rate of electron transport was decreased as the concentration of $Cd^{2+}$ was increased from 1 to $100\;\mu\textrm{M}$. The inhibitory effect of $Cd^{2+}$ was reduced when diphenylcarbazide was added to the reaction medium, indicating that $Cd^{2+}$ affects primarily psn oxygen evolving complexes of thylakoid membrane. The inhibitory effect of $Cd^{2+}$ was reduced when $Mn^{2+}\;and\;Ca^{2+}$ were added to the reaction medium, but the inhibitory effect was not fully relieved. Although the activity of psn was decreased significantly by the treatment of $50\;\mu\textrm{M}\;Cd^{2+}$, Fv/Fm was decreased slightly. However, the treatment of $100\;\mu\textrm{M}\;Cd^{2+}$ resulted in the marked decrease of Fv/Fm. In photosystem I, the rate of electron transport decreased as the concentration of $Cd^{2+}$ was increased from 0.2 to 3.2 mM. The inhibitory effect of $Cd^{2+}$ was decreased when the chloroplast treated with $Cd^{2+}$ was washed by centrifugation.gation.

• Journal of Plant Biology
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• v.35 no.3
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• pp.247-252
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• 1992

The effect of iso-butanol on the electron transport rate of PS I and PS II was investigated in isolated spinach chloroplasts. In photosystem I, the rate of electron transport increased in the presence of 1 to 4% of isobutanol but decreased in 5 to 9% of iso-butanol. But in photosystem II, the rate of electron transport decreased when treated with 0.2 to 1% of iso-butanol. The inhibitory effect of isomers of butanol on PS II electron transport rate increased in the order of 2-butanol, tert-butanol, iso-butanol and I-butanol. This means that PS II activity was affected according to the arrangement of carbon atoms in butanol. The inhibitory effect of iso-butanol reduced when DPC was added in the solution. This means that iso-butanol affects PS II reduction side of thylakoid membrane primarily. The inhibitory effect of iso-butanol was reduced when $Mn^{2+},\;C^{2+}$ or BSA were added in the solution. PS II activity was restored when 1% iso-butanol treated chloroplast solution was diluted to twentyfold or when $Mn^{2+},\;C^{2+}$ or BSA was added to the diluted solution. However, the SDS-PAGE banding pattern of thylakoid membrane proteins was similar even in 2% iso-butanol treated chloroplasts and the control ones. Only in 5% iso-butanol treated chloroplasts these bands were very weak. These observations suggest that low concentrations of iso-butanol releases manganese and calcium ions from chloroplasts and inhibits the electron transport system. This inhibitory effect can be reversible in low concenterations but in high concentrations the inhibitory effect of iso-butanol become irreversible.rsible.

• Korean Journal of Weed Science
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• v.14 no.1
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• pp.1-7
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• 1994

The effects of S-23142{N-(4-chloro-2-fluoro-5-propargyloxyphenyl)-3, 4, 5, 6-tetrahydrophtalimide}, on protoporphyrin IX(PPIX) biosynthesis in Spinacia oleracea L, leaf in vivo and in vitro condition were investigated by reversed-phase HPLC with fluorescence detector. The stroma and the membrane fraction of spinach chloroplast were isolated by osmotic regulation. The conversion of ${\delta}$-aminolevulinic acid(ALA) to PPIX occured more in the stroma than in the membrane fraction. It suggested that the enzymes that catalyse PPIX biosynthesis from ALA were localized in the stroma. Also, the synthesized PPIX content from ALA was completely inhibited by $10^{-8}M$ of S-23412 or $10^{-7}M$ of acifluorfen in the stroma but not in the membrane fractions. Therefore, these results suggested that the target site of S-23142 and acifluorfen may exist in the stroma fraction of spinach chloroplast.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.39 no.6
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• pp.556-563
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• 1994

To compare the tolerance of crops to acid rain of simulated acid rains(SAR) of three pH levels (3.0, 4.5, and 6.0) were applied to rice, corn, soybean, adzuki bean, hot pepper, tomato, sesame, barley, wheat, Chinese cabbage, radish, lettuce, and spinach for 30 days from 20 days after emergence at two-day interval with 10mm at a time. No visual damages were observed. Also, no alteration of ultrastructure was observed although some lightly stained lipid granules were observed in the chloroplasts of sesame, soybean, and adzuki bean at the SAR treatment of pH 3.0. As pH of SAR decreased, chlorophyll content increased in adzuki bean, decreased in rice, tomato, and spinach, and similar in the other crops. Photosynthetic activity of adzuki bean increased, while decreased in Chinese cabbage and barley as pH of SAR decreased, and similar in other crops. Concentration and uptake of N were not affected by SAR treatments in all crops except tomato and barley. When a strong SAR of pH 2.0 was applied, rice, corn, sesame, tomato, barley, and wheat were relatively tolerant, while adzuki bean, hot pepper, soybean, Chinese cabbage, lettuce, radish, and spinach were susceptible in terms of visual damages.

• Journal of Plant Biology
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• v.36 no.4
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• pp.313-319
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• 1993

Photoproduction of hydrogen by free and polyvinylalcohol (PVA)-immobilized spinach chloroplasts was investigated. Immobilization of chloroplast with PVA increased the functional stability of the chloroplast during storage. PVA-immobilized chloroplasts preserved photosynthetic electron transport activity much better than free chloroplasts. The hydrogen production of free chloroplast decreased to 17% of initial activity after storage of six days. The hydrogen production of the PVA-immobilized chloroplast, however, showed 44% of initial activity after storage of 15 days. The maximal rate of hydrogen production was accomplished at 2$^{\circ}C$ under the light intensity above 116 $\mu$E.m-2.s-1. The amount of hydrogen produced was proportional to the chlorophyll concentration. The hydrogen production was inhibited by DCMU treatment, indicating hydrogen production is dependent on photosynthetic electron transport. These results suggest that PVA is a good candidate for the immobilization matrix of chloroplasts for the photoproduction of hydrogen.

• Journal of Plant Biology
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• v.34 no.3
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• pp.215-221
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• 1991

Photoxynthetic properties of polyvinylalcohol (PVA)-immobilized chloroplast especially regarded to stability of photosynthetic electron transport and the fluorescence induction pattern were studied. When isolated spinach chloroplasts were immobilized with PVA, it showed good preservation of photosynthetic electron transport activity, especially PS II activity, during storage at -15$^{\circ}C$, 4$^{\circ}C$ and 2$0^{\circ}C$. And immobilized chloroplasts revealed similar thermostability of whole chain electron transport to free chloroplsts. And the absorption peak of red band of chloroplasts showed the blue-shift of 2-4 nm after immobilization. Fv/Fm ratio of chlorophyll fluorescence slightly decreased after immobilization. White light pulse after continuous light do not induced the additional fluorescence rise. This means chlorophyll fluorescence at room temperature reached to Fmax under continuous light in the immobilized chloroplasts. It seems that PVA may be a good candidate for immobilization matrix for the preservation of photosynthetic function of thylakoids and for the continuous use of chloroplast membranes of higher plants for solar energy storage and conversion.

• Journal of Plant Biology
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• v.19 no.3
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• pp.71-84
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• 1976

In order to observe the phosphate metabolism in chloroplast, the contents of inorganic phosphate and various compounds in chloroplast from spinach leaf tissues were investigated during the reaction in the light and dark in the reaction mixture and the turnover of phosphate in chloroplast was compared with that of whole cell system: 1. The phosphorus of DNA in chloroplast appears to be transferred from inorganic phosphate, while in whole cell system from phosphate pool. 2. $^{32}P-phosphate$ content of acid soluble fraction in chloroplast as well as in whole cell system was more increased in the light than dark during the reaction. It was noted to be caused by the stimulation of sugar phosphate synthesis in the light. 3. It was confirmed that polyphosphate exists in chloroplast as well as whole cell. Acid insoluble polyphosphate content in whole cell system was significantly decreased during the reaction and the similar tendency was also observed in chloroplst. It is, therefore, considered that acid insoluble polyphosphate also play an most important role as a phosphate pool respectively in chloroplast and in cytoplasm. 4. Protein and lipid phosphorus in chloroplast as well as whole cell system were transferred from acid insoluble polyphosphate.

• Journal of Plant Biology
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• v.19 no.4
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• pp.95-99
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• 1976

This work deals with different extraction media and reaction mixtures on photosystem II activity of Spinach chloroplasts. The photoreduction rate of ferricyanide and DPIP by intact chloroplasts which extracted with four kinds of extraction media; S-Tris-N pH 7.2, 8.0, S-Tricine-N pH 7.2, 8.0, was measured in five kinds of reaction mixtures; S-Tris-N pH 7.2, 8.0, S-Tricine-N pH 7.2, 8.0, 0.05 M-Tris pH 7.2. The extraction medium which shows the highest photoreduction rate was S-Tris-N at pH 7.2 and S-Tricine-N at pH 8.0. As to the reaciton mixture, S-Tricine-N pH 8.0 showed the highest rate. On the complex effects of extraction media and reaction mixtures, the highest photordeuction rate of Hill oxidant by intact chloroplasts was obtained by S-Tris-N pH 7.2 extraction medium and S-Tricine-N pH 8.0 reaction mixture. The second highest activity was obtained by S-Tricine-N pH 8.0 extraction medium and reaction mixture.

• Journal of Plant Biology
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• v.33 no.4
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• pp.217-223
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• 1990

Restriction enzyme inhibiting substance (REIS) extracted from spinach chloroplast and Chlamydomonas seems not to be proteinaceous, because its inhibiting activity was not lost by heat or trypsin treatment. And it seems not to be lipid or polysacchride, because its inhibiting activity was not lost by lipase or $\alpha$-amylase treatment, respectively. In Chlamydomonas, putrescine, spermidine and spermine were present. The amount of putrescine was the smallest and that of spermie was the greatest. But only spermine was contained in REIS and the activity of REIS. It was proportional to the amount of spermine in REIS and it was hindered by Na+ ion. So, the inhibiting activity of REIS seems to be deeply related to spermine contained in REIS. But restriction enzyme inhibiting activity remained to the some extent although salts and spermine were eliminated by dialysis.