和田 慎也 | ![]() |
ワダ シンヤ | |
大学院農学研究科 生命機能科学専攻 | |
助教 | |
農芸化学関係 |
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研究論文(学術雑誌)
PSI photoinhibition is usually avoided through P700 oxidation. Without this protective mechanism, excess light represents a potentially lethal threat to plants. PGR5 is suggested to be a major component of cyclic electron transport around PSI and is important for P700 oxidation in angiosperms. The known Arabidopsis PGR5 deficient mutant, pgr5-1, is incapable of P700 oxidation regulation and has been used in numerous photosynthetic studies. However, here it was revealed that pgr5-1 was a double mutant with exaggerated PSI photoinhibition. pgr5-1 significantly reduced growth compared to the newly isolated PGR5 deficient mutant, pgr5hope1. The introduction of PGR5 into pgr5-1 restored P700 oxidation regulation, but remained a pale-green phenotype, indicating that pgr5-1 had additional mutations. Both pgr5-1 and pgr5hope1 tended to cause PSI photoinhibition by excess light, but pgr5-1 exhibited an enhanced reduction in PSI activity. Introducing AT2G17240, a candidate gene for the second mutation into pgr5-1 restored the pale-green phenotype and partially restored PSI activity. Furthermore, a deficient mutant of PGRL1 complexing with PGR5 significantly reduced PSI activity in the double-deficient mutant with AT2G17240. From these results, we concluded that AT2G17240, named PSI photoprotection 1 (PTP1), played a role in PSI photoprotection, especially in PGR5/PGRL1 deficient mutants.
MDPI AG, 2021年10月26日, Cells, 10 (11), 2884 - 2884[査読有り]
研究論文(学術雑誌)
In response to decreases in the assimilation efficiency of CO2, plants oxidize the reaction center chlorophyll (P700) of photosystem I (PSI) to suppress reactive oxygen species (ROS) production. In hydro-cultured sunflower leaves experiencing essential mineral deficiencies, we analyzed the following parameters that characterize PSI and PSII: (1) the reduction-oxidation states of P700 [Y(I), Y(NA), and Y(ND)]; (2) the relative electron flux in PSII [Y(II)]; (3) the reduction state of the primary electron acceptor in PSII, QA (1 − qL); and (4) the non-photochemical quenching of chlorophyll fluorescence (NPQ). Deficiency treatments for the minerals N, P, Mn, Mg, S, and Zn decreased Y(II) with an increase in the oxidized P700 [Y(ND)], while deficiencies for the minerals K, Fe, Ca, B, and Mo decreased Y(II) without an increase in Y(ND). During the induction of photosynthesis, the above parameters showed specific responses to each mineral. That is, we could diagnose the mineral deficiency and identify which mineral affected the photosynthesis parameters.
MDPI AG, 2021年06月23日, Antioxidants, 10 (7), 996 - 996研究論文(学術雑誌)
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Upon exposure to environmental stress, the primary electron donor in photosystem I (PSI), P700, is oxidized to suppress the production of reactive oxygen species that could oxidatively inactivate the function of PSI. The illumination of rice leaves with actinic light induces intrinsic fluctuations in the opening and closing of stomata, causing the net CO2 assimilation rate to fluctuate. We examined the effects of these intrinsic fluctuations on electron transport reactions. Under atmospheric O2 conditions (21 kPa), the effective quantum yield of photosystem II (PSII) (Y(II)) remained relatively high while the net CO2 assimilation rate fluctuated, which indicates the function of alternative electron flow. By contrast, under low O2 conditions (2 kPa), Y(II) fluctuated. These results suggest that photorespiration primarily drove the alternative electron flow. Photorespiration maintained the oxidation level of ferredoxin (Fd) throughout the fluctuation of the net CO2 assimilation rate. Moreover, the relative activity of photorespiration was correlated with both the oxidation level of P700 and the magnitude of the proton gradient across the thylakoid membrane in 21 kPa O2 conditions. These results show that photorespiration oxidized P700 by stimulating the proton gradient formation when CO2 assimilation was suppressed by stomatal closure.
MDPI AG, 2020年12月12日, Plants, 9 (12), 1761 - 1761研究論文(学術雑誌)
It is of interest how photosynthetic electron transport (PET) reactions respond to excess light energy caused by the combination of drought stress and high temperatures. Since such information is scarcely available for photosystem I (PSI), this question was explored in rice (Oryza sativa L.) plants subjected to drought stress, using culture solutions that contain poly(ethylene glycol) at different concentrations under two day/night temperature regimes. At 27/22 °C (day/night), drought stress led to the oxidation of the reaction center of the chlorophyll of PSI (P700), and also led to decreases in the quantum efficiencies of photosystem II (PSII) and PSI, and a reduction of the primary quinone electron acceptor of PSI. Such drought stress responses were wholly stimulated at 35/30 °C. These parameters were strongly correlated with each other and were minimally affected by temperature. These results indicate that the drought stress responses of the respective PET reactions are closely associated with each other in the oxidization of P700 and that such responses are stimulated at high temperatures. The underlying mechanisms of these phenomena were discussed. While P700 oxidation is thought to suppress reactive oxygen species (ROS) production, PSI photoinhibition was observed under severe stress conditions, implying that P700 oxidation is not sufficient for the protection of PSI under drought stress.
MDPI AG, 2019年04月26日, International Journal of Molecular Sciences, 20 (9), 2068 - 2068[査読有り]
研究論文(学術雑誌)
Overproduction of Rubisco did not proportionately increase the rate of CO2 assimilation (A) in rice probably because the capacity of regeneration of the substrate of Rubisco, ribulose-1,5-bisphosphate (RuBP), could not afford the increased Rubisco capacity. Since sedoheptulose-1,7-bisphosphatase (SBPase) is thought to be one of the limiting factors of RuBP regeneration capacity, SBPase and Rubisco were co-overproduced in rice in order to improve photosynthesis. Although SBPase and Rubisco contents increased by 82-102% and 20-30%, respectively, A did not increase under the conditions of high irradiance and different [CO2] partial pressures. Thus, co-overproduction of SBPase and Rubisco did not improve photosynthesis in rice. Overproduction and antisense suppression of SBPase did not greatly affect A at high [CO2], although it is thought to be determined by RuBP regeneration capacity. These results strongly suggest that SBPase does not determine RuBP regeneration capacity in rice. This is likely to be the reason for the unimproved photosynthesis by co-overproduction of SBPase and Rubisco.
TAYLOR & FRANCIS LTD, 2019年01月02日, SOIL SCIENCE AND PLANT NUTRITION, 65 (1), 36 - 40, 英語[査読有り]
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Flavodiiron protein (FLV) mediates photoreduction of O2to H2O. It is conserved from cyanobacteria to gymnosperms but not in angiosperms. The introduction of a moss (Physcomitrella patens) FLV (PpFLV) gene into Arabidopsis (Arabidopsis thaliana) made photosystem I (PSI) resistant to fluctuating light. Here, we used the same strategy with three rice (Oryza sativa) genotypes. PpFLV in the wild-type rice background functioned as an efficient PSI electron sink and increased resistance to PSI photodamage under fluctuating light. The introduction of PpFLV into the PGR5-RNAi mutant [defective in PROTON GRADIENT REGULATION5 (PGR5)-dependent cyclic electron transport around PSI, CET-PSI], the crr6 mutant [defective in chloroplast NAD(P)H-dehydrogenase-like complex (NDH)-dependent CET-PSI], and the PGR5-RNAi crr6 double mutant (double defective in CET-PSI activity) alleviated PSI photodamage under fluctuating light. Furthermore, PpFLV substituted for the function of PGR5- and NDH-dependent CET-PSI without competing for CO2 assimilation under constant light, as there was no difference in CO2 assimilation per Rubisco content and biomass production was recovered to the wild-type level. Thus, the exogenous FLV system could act not only as a safety valve under fluctuating light, but also generate a proton motive force for balancing the ATP/NADPH production ratio during steady-state photosynthesis.
American Society of Plant Biologists, 2018年02月01日, Plant Physiology, 176 (2), 1509 - 1518, 英語[査読有り]
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Chloroplasts are the primary energy suppliers for plants, and much of the total leaf nitrogen is distributed to these organelles. During growth and reproduction, chloroplasts in turn represent a major source of nitrogen to be recovered from senescing leaves and used in newly-forming and storage organs. Chloroplast proteins also can be an alternative substrate for respiration under suboptimal conditions. Autophagy is a process of bulk degradation and nutrient sequestration that is conserved in all eukaryotes. Autophagy can selectively target chloroplasts as whole organelles and or as Rubisco-containing bodies that are enclosed by the envelope and specifically contain the stromal portion of the chloroplast. Although information is still limited, recent work indicates that chloroplast recycling via autophagy plays important roles not only in developmental processes but also in organelle quality control and adaptation to changing environments. This article is part of a Special Issue entitled: Dynamic and ultrastructure of bioenergetic membranes and their components. (C) 2013 Elsevier B.V. All rights reserved.
ELSEVIER SCIENCE BV, 2014年04月, BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS, 1837 (4), 512 - 521, 英語[査読有り]
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論文集(書籍)内論文
Oxygen (O2)-evolving photosynthetic organisms oxidize the reaction center chlorophyll, P700, in photosystem I (PSI) to suppress the production of reactive oxygen species. The oxidation of P700 is accompanied by alternative electron flow in PSI (AEF-I), which is not required for photosynthetic linear electron flow (LEF). To characterize AEF-I, we compared the redox reactions of P700 and ferredoxin (Fd) during the induction of carbon dioxide (CO2) assimilation in wheat leaves, using dark-interval relaxation kinetics analysis. Switching on an actinic light (1000 μmol photons m-2 s-1) at ambient CO2 partial pressure of 40 Pa and ambient O2 partial pressure of 21 kPa gradually oxidized P700 (P700+) and enhanced the reduction rate of P700+ (vP700) and oxidation rate of reduced Fd (vFd). The vFd showed a positive linear relationship with an apparent photosynthetic quantum yield of PSII (Y[II]) originating at point zero; the redox turnover of Fd is regulated by LEF via CO2 assimilation and photorespiration. The vP700 also showed a positive linear relationship with Y(II), but the intercept was positive, not zero. That is, the electron flux in PSI included the electron flux in AEF-I in addition to that in LEF. This indicates that the oxidation of P700 induces AEF-I. We propose a possible mechanism underlying AEF-I and its physiological role in the mitigation of oxidative damage.
2019年06月05日, Plants (Basel, Switzerland), 8 (6), 英語, 国際誌研究論文(学術雑誌)
Photorespiration coupled with CO2 assimilation is thought to act as a defense system against photoinhibition caused by osmotic stress. In the present study, we examined whether such a mechanism is operative for the protection of photosystem I (PSI) in rice (Oryza sativa L.) including transgenic plants with decreased and increased Rubisco content (RBCS-antisense and RBCS-sense plants, respectively). All plants were hydroponically grown and moderate osmotic stress was imposed using hydroponic culture solutions containing poly(ethylene glycol) (PEG) at 16% or 20% (w/v) for 2 d. In wild-type plants, the rates of CO2 assimilation (A) were significantly decreased by the PEG treatment, whereas the photorespiration activity estimated from the rates of electron transport in photosystem II (PSII) and A were not affected. The maximal quantum efficiency of PSII (Fv/Fm) and the maximal activity of PSI (Pm) were also not affected. In RBCS-antisense plants, A and the estimated photorespiration activity were considerably lower than those in wild-type plants in the presence or absence of the PEG treatment. Pm and both Fv/Fm and Pm decreased in the 16% PEG-treated and 20% PEG-treated RBCS-antisense plants, respectively. Thus, the decrease in Rubisco content led to the photoinhibition of PSI and PSII, indicating the importance of photorespiration coupled with CO2 assimilation for the protection of PSI from moderate PEG-induced osmotic stress. It was also shown that PSI was more sensitive to osmotic stress than PSII. In the PEG-treated wild-type and RBCS-antisense plants, osmotic-stress responses of the photosynthetic electron transport reactions upstream of PSI led to the oxidation of P700, which is thought to prevent PSI from over-reduction. Although such a defense system operated, it was not sufficient for the protection of PSI in RBCS-antisense plants. In addition, there were no large differences in the parameters measured between wild-type and RBCS-sense plants, as overproduction of Rubisco did not increase photorespiration activity.
2020年, Frontiers in plant science, 11, 1121 - 1121, 英語, 国際誌研究論文(学術雑誌)
Chloroplasts contain approximately 80% of total leaf nitrogen and represent a major source of recycled nitrogen during leaf senescence. While bulk degradation of the cytosol and organelles in plants is mediated by autophagy, its role in chloroplast catabolism is largely unknown. We investigated the effects of autophagy disruption on the number and size of chloroplasts during senescence. When leaves were individually darkened, senescence was promoted similarly in both wild-type Arabidopsis (Arabidopsis thaliana) and in an autophagy-defective mutant, atg4a4b-1. The number and size of chloroplasts decreased in darkened leaves of wild type, while the number remained constant and the size decrease was suppressed in atg4a4b-1. When leaves of transgenic plants expressing stroma-targeted DsRed were individually darkened, a large accumulation of fluorescence in the vacuolar lumen was observed. Chloroplasts exhibiting chlorophyll fluorescence, as well as Rubisco-containing bodies, were also observed in the vacuole. No accumulation of stroma-targeted DsRed, chloroplasts, or Rubisco-containing bodies was observed in the vacuoles of the autophagy-defective mutant. We have succeeded in demonstrating chloroplast autophagy in living cells and provide direct evidence of chloroplast transportation into the vacuole.
AMER SOC PLANT BIOLOGISTS, 2009年02月, PLANT PHYSIOLOGY, 149 (2), 885 - 893, 英語[査読有り]
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Autophagy is an intracellular process facilitating the vacuolar degradation of cytoplasmic components and is important for nutrient recycling during starvation. We previously demonstrated that chloroplasts can be partially mobilized to the vacuole by autophagy via spherical bodies named Rubisco-containing bodies (RCBs). Although chloroplasts contain approximately 80% of total leaf nitrogen and represent a major carbon and nitrogen source for new growth, the relationship between leaf nutrient status and RCB production remains unclear. We examined the effects of nutrient factors on the appearance of RCBs in leaves of transgenic Arabidopsis (Arabidopsis thaliana) expressing stroma-targeted fluorescent proteins. In excised leaves, the appearance of RCBs was suppressed by the presence of metabolic sugars, which were added externally or were produced during photosynthesis in the light. The light-mediated suppression was relieved by the inhibition of photosynthesis. During a diurnal cycle, RCB production was suppressed in leaves excised at the end of the day with high starch content. Starchless mutants phosphoglucomutase and ADP-Glc pyrophosphorylase1 produced a large number of RCBs, while starch-excess mutants starch-excess1 and maltose-excess1 produced fewer RCBs. In nitrogen-limited plants, as leaf carbohydrates were accumulated, RCB production was suppressed. We propose that there exists a close relationship between the degradation of chloroplast proteins via RCBs and leaf carbon but not nitrogen status in autophagy. We also found that the appearance of non-RCB-type autophagic bodies was not suppressed in the light and somewhat responded to nitrogen in excised leaves, unlike RCBs. These results imply that the degradation of chloroplast proteins via RCBs is specifically controlled in autophagy.
AMER SOC PLANT BIOLOGISTS, 2010年11月, PLANT PHYSIOLOGY, 154 (3), 1196 - 1209, 英語[査読有り]
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