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Dive into the research topics where Daisuke Takagi is active.

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Featured researches published by Daisuke Takagi.


Plant Physiology | 2016

Superoxide and singlet oxygen produced within the thylakoid membranes both cause photosystem I photoinhibition

Daisuke Takagi; Shigeo Takumi; Masaki Hashiguchi; Takehiro Sejima; Chikahiro Miyake

Repetitive short-pulse illumination produces both superoxide and singlet oxygen within the thylakoid membranes, leading to inactivation of photosystem I. Photosystem I (PSI) photoinhibition suppresses plant photosynthesis and growth. However, the mechanism underlying PSI photoinhibition has not been fully clarified. In this study, in order to investigate the mechanism of PSI photoinhibition in higher plants, we applied repetitive short-pulse (rSP) illumination, which causes PSI-specific photoinhibition in chloroplasts isolated from spinach leaves. We found that rSP treatment caused PSI photoinhibition, but not PSII photoinhibition in isolated chloroplasts in the presence of O2. However, chloroplastic superoxide dismutase and ascorbate peroxidase activities failed to protect PSI from its photoinhibition. Importantly, PSI photoinhibition was largely alleviated in the presence of methyl viologen, which stimulates the production of reactive oxygen species (ROS) at the stromal region by accepting electrons from PSI, even under the conditions where CuZn-superoxide dismutase and ascorbate peroxidase activities were inactivated by KCN. These results suggest that the ROS production site, but not the ROS production rate, is critical for PSI photoinhibition. Furthermore, we found that not only superoxide (O2−) but also singlet oxygen (1O2) is involved in PSI photoinhibition induced by rSP treatment. From these results, we suggest that PSI photoinhibition is caused by both O2− and 1O2 produced within the thylakoid membranes when electron carriers in PSI become highly reduced. Here, we show, to our knowledge, new insight into the PSI photoinhibition in higher plants.


Plant and Cell Physiology | 2014

Repetitive Short-Pulse Light Mainly Inactivates Photosystem I in Sunflower Leaves

Takehiro Sejima; Daisuke Takagi; Hiroshi Fukayama; Amane Makino; Chikahiro Miyake

Under field conditions, the leaves of plants are exposed to fluctuating light, as observed in sunfleck. The duration and frequency of sunfleck, which is caused by the canopy being blown by the wind, are in the ranges from 0.2 to 50 s, and from 0.004 to 1 Hz, respectively. Furthermore, >60% of the sunfleck duration ranges from 0.2 to 0.8 s. In the present research, we analyzed the effects of repetitive illumination by short-pulse (SP) light of sunflower leaves on the photosynthetic electron flow. The duration of SP light was set in the range from 10 to 300 ms. We found that repetitive illumination with SP light did not induce the oxidation of P700 in PSI, and mainly inactivated PSI. Increases in the intensity, duration and frequency of SP light enhanced PSI photoinhibition. PSI photoinhibition required the presence of O2. The inactivation of PSI suppressed the net CO2 assimilation. On the other hand, the increase in the oxidized state of P700 suppressed PSI inactivation. That is, PSI with a reduced reaction center would produce reactive oxygen species (ROS) by SP light, leading to PSI photodamage. This mechanism probably explains the PSI photodamage induced by constant light.


Plant and Cell Physiology | 2014

The Calvin Cycle Inevitably Produces Sugar-Derived Reactive Carbonyl Methylglyoxal During Photosynthesis: A Potential Cause of Plant Diabetes

Daisuke Takagi; Hironori Inoue; Mizue Odawara; Ginga Shimakawa; Chikahiro Miyake

Sugar-derived reactive carbonyls (RCs), including methylglyoxal (MG), are aggressive by-products of oxidative stress known to impair the functions of multiple proteins. These advanced glycation end-products accumulate in patients with diabetes mellitus and cause major complications, including arteriosclerosis and cardiac insufficiency. In the glycolytic pathway, the equilibration reactions between dihydroxyacetone phosphate and glyceraldehyde 3-phosphate (GAP) have recently been shown to generate MG as a by-product. Because plants produce vast amounts of sugars and support the same reaction in the Calvin cycle, we hypothesized that MG also accumulates in chloroplasts. Incubating isolated chloroplasts with excess 3-phosphoglycerate (3-PGA) as the GAP precursor drove the equilibration reaction toward MG production. The rate of oxygen (O2) evolution was used as an index of 3-PGA-mediated photosynthesis. The 3-PGA- and time-dependent accumulation of MG in chloroplasts was confirmed by HPLC. In addition, MG production increased with an increase in light intensity. We also observed a positive linear relationship between the rates of MG production and O2 evolution (R = 0.88; P < 0.0001). These data provide evidence that MG is produced by the Calvin cycle and that sugar-derived RC production is inevitable during photosynthesis. Furthermore, we found that MG production is enhanced under high-CO2 conditions in illuminated wheat leaves.


Physiologia Plantarum | 2017

Diversity of strategies for escaping reactive oxygen species production within photosystem I among land plants: P700 oxidation system is prerequisite for alleviating photoinhibition in photosystem I

Daisuke Takagi; Kimitsune Ishizaki; Hitomi Hanawa; Tomohito Mabuchi; Ginga Shimakawa; Hiroshi Yamamoto; Chikahiro Miyake

In land plants, photosystem I (PSI) photoinhibition limits carbon fixation and causes growth defects. In addition, recovery from PSI photoinhibition takes much longer than PSII photoinhibition when the PSI core-complex is degraded by oxidative damage. Accordingly, PSI photoinhibition should be avoided in land plants, and land plants should have evolved mechanisms to prevent PSI photoinhibition. However, such protection mechanisms have not yet been identified, and it remains unclear whether all land plants suffer from PSI photoinhibition in the same way. In the present study, we focused on the susceptibility of PSI to photoinhibition and investigated whether mechanisms of preventing PSI photoinhibition varied among land plant species. To assess the susceptibility of PSI to photoinhibition, we used repetitive short-pulse (rSP) illumination, which specifically induces PSI photoinhibition. Subsequently, we found that land plants possess a wide variety of tolerance mechanisms against PSI photoinhibition. In particular, gymnosperms, ferns and mosses/liverworts exhibited higher tolerance to rSP illumination-induced PSI photoinhibition than angiosperms, and detailed analyses indicated that the tolerance of these groups could be partly attributed to flavodiiron proteins, which protected PSI from photoinhibition by oxidizing the PSI reaction center chlorophyll (P700) as an electron acceptor. Furthermore, we demonstrate, for the first time, that gymnosperms, ferns and mosses/liverworts possess a protection mechanism against photoinhibition of PSI that differs from that of angiosperms.


Photosynthesis Research | 2016

Photorespiration provides the chance of cyclic electron flow to operate for the redox-regulation of P700 in photosynthetic electron transport system of sunflower leaves

Daisuke Takagi; Masaki Hashiguchi; Takehiro Sejima; Amane Makino; Chikahiro Miyake

To elucidate the molecular mechanism to oxidize the reaction center chlorophyll, P700, in PSI, we researched the effects of partial pressure of O2 (pO2) on photosynthetic characteristic parameters in sunflower (Helianthus annuusxa0L.) leaves. Under low CO2 conditions, the oxidation of P700 was stimulated; however the decrease in pO2 suppressed its oxidation. Electron fluxes in PSII [Y(II)] and PSI [Y(I)] showed pO2-dependence at low CO2 conditions. H+-consumption rate, estimated from Y(II) and CO2-fixation/photorespiration rates (JgH+), showed the positive curvature relationship with the dissipation rate of electrochromic shift signal (VH+), which indicates H+-efflux rate from lumen to stroma in chloroplasts. Therefore, these electron fluxes contained, besides CO2-fixation/photorespiration-dependent electron fluxes, non-H+-consumption electron fluxes including Mehler-ascorbate peroxidase (MAP)-pathway. Y(I) that was larger than Y(II) surely implies the functioning of cyclic electron flow (CEF). Both MAP-pathway and CEF were suppressed at lower pO2, with plastoquinone-pool reduced. That is, photorespiration prepares the redox-poise of photosynthetic electron transport system for CEF activity as an electron sink. Excess Y(II), [ΔY(II)] giving the curvature relationship with VH+, and excess Y(I) [ΔCEF] giving the difference between Y(I) and Y(II) were used as an indicator of MAP-pathway and CEF activity, respectively. Although ΔY(II) was negligible and did not show positive relationship to the oxidation-state of P700, ΔCEF showed positive linear relationship to the oxidation-state of P700. These facts indicate that CEF cooperatively with photorespiration regulates the redox-state of P700 to suppress the over-reduction in PSI under environmental stress conditions.


Plant Journal | 2017

Chloroplastic ATP synthase builds up a proton motive force preventing production of reactive oxygen species in photosystem I

Daisuke Takagi; Katsumi Amako; Masaki Hashiguchi; Hidehiro Fukaki; Kimitsune Ishizaki; Tatsuaki Goh; Yoichiro Fukao; Ryosuke Sano; Tetsuya Kurata; Taku Demura; Shinichiro Sawa; Chikahiro Miyake

Over-reduction of the photosynthetic electron transport (PET) chain should be avoided, because the accumulation of reducing electron carriers produces reactive oxygen species (ROS) within photosystem I (PSI) in thylakoid membranes and causes oxidative damage to chloroplasts. To prevent production of ROS in thylakoid membranes the H+ gradient (ΔpH) needs to be built up across the thylakoid membranes to suppress the over-reduction state of the PET chain. In this study, we aimed to identify the critical component that stimulates ΔpH formation under illumination in higher plants. To do this, we screened ethyl methane sulfonate (EMS)-treated Arabidopsis thaliana, in which the formation of ΔpH is impaired and the PET chain caused over-reduction under illumination. Subsequently, we isolated an allelic mutant that carries a missense mutation in the γ-subunit of chloroplastic CF0 CF1 -ATP synthase, named hope2. We found that hope2 suppressed the formation of ΔpH during photosynthesis because of the high H+ efflux activity from the lumenal to stromal side of the thylakoid membranes via CF0 CF1 -ATP synthase. Furthermore, PSI was in a more reduced state in hope2 than in wild-type (WT) plants, and hope2 was more vulnerable to PSI photoinhibition than WT under illumination. These results suggested that chloroplastic CF0 CF1 -ATP synthase adjusts the redox state of the PET chain, especially for PSI, by modulating H+ efflux activity across the thylakoid membranes. Our findings suggest the importance of the buildup of ΔpH depending on CF0 CF1 -ATP synthase to adjust the redox state of the reaction center chlorophyll P700 in PSI and to suppress the production of ROS in PSI during photosynthesis.


Physiologia Plantarum | 2016

Post-illumination transient O2-uptake is driven by photorespiration in tobacco leaves

Takehiro Sejima; Hitomi Hanawa; Ginga Shimakawa; Daisuke Takagi; Yuji Suzuki; Hiroshi Fukayama; Amane Makino; Chikahiro Miyake

This study aims to elucidate the molecular mechanism for the transient increase in the O2 -uptake rate in tobacco (Nicotiana tabacum cv Xanthi) leaves after turning off actinic lights (ALs). The photosynthetic O2 evolution rate reaches a maximum shortly after the onset of illumination with ALs and then decreases to zero in atmospheric CO2 /O2 conditions. After turning off the ALs, tobacco leaves show a transient increase in the O2 -uptake rate, the post-illumination transient O2 -uptake, and thereafter, the O2 -uptake rate decreases to the level of the dark-respiration rate. Photosynthetic linear electron flow, evaluated as the quantum yield of photosystem II [Y(II)], maintained a steady-state value distinct from the photosynthetic O2 -evolution rate. In high-[CO2 ] conditions, the photosynthetic O2 -evolution rate and Y(II) showed a parallel behavior, and the post-illumination transient O2 -uptake was suppressed. On the other hand, in maize leaves (a C4 plant), even in atmospheric CO2 /O2 conditions, Y(II) paralleled the photosynthetic O2 -evolution rate and the post-illumination transient O2 -uptake was suppressed. Hypothesizing that the post-illumination transient O2 -uptake is driven by C3 plant photorespiration in tobacco leaves, we calculated both the ribulose 1,5-bisphosphate carboxylase- and oxygenase-rates (Vc and Vo) from photosynthetic O2 -evolution and the post-illumination transient O2 -uptake rates. These values corresponded to those estimated from simultaneous chlorophyll fluorescence/O2 -exchange analysis. Furthermore, the H+ -consumption rate for ATP synthesis in both photosynthesis and photorespiration, calculated from both Vc and Vo that were estimated from chlorophyll fluorescence/CO2 -exchange analysis, showed a positive linear relationship with the dissipation rate of the electrochromic shift signal. Thus, these findings support our hypothesis.


Physiologia Plantarum | 2017

Land plants drive photorespiration as higher electron‐sink: comparative study of post‐illumination transient O2‐uptake rates from liverworts to angiosperms through ferns and gymnosperms

Hitomi Hanawa; Kimitsune Ishizaki; Kana Nohira; Daisuke Takagi; Ginga Shimakawa; Takehiro Sejima; Keiichiro Shaku; Amane Makino; Chikahiro Miyake

In higher plants, the electron-sink capacity of photorespiration contributes to alleviation of photoinhibition by dissipating excess energy under conditions when photosynthesis is limited. We addressed the question at which point in the evolution of photosynthetic organisms photorespiration began to function as electron sink and replaced the flavodiiron proteins which catalyze the reduction of O2 at photosystem I in cyanobacteria. Algae do not have a higher activity of photorespiration when CO2 assimilation is limited, and it can therefore not act as an electron sink. Using land plants (liverworts, ferns, gymnosperms, and angiosperms) we compared photorespiration activity and estimated the electron flux driven by photorespiration to evaluate its electron-sink capacity at CO2 -compensation point. In vivo photorespiration activity was estimated by the simultaneous measurement of O2 -exchange rate and chlorophyll fluorescence yield. All C3-plants leaves showed transient O2 -uptake after actinic light illumination (post-illumination transient O2 -uptake), which reflects photorespiration activity. Post-illumination transient O2 -uptake rates increased in the order from liverworts to angiosperms through ferns and gymnosperms. Furthermore, photorespiration-dependent electron flux in photosynthetic linear electron flow was estimated from post-illumination transient O2 -uptake rate and compared with the electron flux in photosynthetic linear electron flow in order to evaluate the electron-sink capacity of photorespiration. The electron-sink capacity at the CO2 -compensation point also increased in the above order. In gymnosperms photorespiration was determined to be the main electron-sink. C3-C4 intermediate species of Flaveria plants showed photorespiration activity, which intermediate between that of C3- and C4-flaveria species. These results indicate that in the first land plants, liverworts, photorespiration started to function as electron sink. According to our hypothesis, the dramatic increase in partial pressure of O2 in the atmosphere about 0.4u2009billion years ago made it possible to drive photorespiration with higher activity in liverworts.


Soil Science and Plant Nutrition | 2012

O2 supports 3-phosphoglycerate-dependent O2 evolution in chloroplasts from spinach leaves

Daisuke Takagi; Hiroshi Yamamoto; Katsumi Amako; Amane Makino; Toshio Sugimoto; Chikahiro Miyake

We tested the hypothesis that the Mehler-ascorbate peroxidase (MAP) pathway supports 3-phosphoglycerate (PGA)-dependent oxygen (O2) evolution using intact chloroplasts. Lowering O2 concentration (<1u2009µM) suppressed PGA-dependent O2 evolution rate. High O2 concentration (about 250u2009µM) enhanced the electron fluxes in Photosystem II (PSII). Also, high O2 concentration oxidized both QA in PSII and Cyt f in thylakoid membranes. These results indicated that the MAP pathway stimulated photosynthetic electron transport. Furthermore, electrochromic shift signal was also increased at high O2 concentration, compared to low O2 concentration. Non-photochemical quenching of chlorophyll fluorescence was also enhanced at high O2 concentration. These data support our hypothesis that the MAP pathway functioned in intact chloroplasts and accelerated PGA-dependent O2 evolution by inducing ΔpH formation to produce and supply adenosine triphosphate (ATP) to the conversion reaction of PGA to glyceraldehyde 3-phosphate through 1,3-diphosphoglycerate in chloroplasts.


Plant Physiology | 2016

Suppression of Chloroplastic Alkenal/One Oxidoreductase Represses the Carbon Catabolic Pathway in Arabidopsis Leaves during Night

Daisuke Takagi; Kentaro Ifuku; Kenichi Ikeda; Kanako Inoue; Pyoyun Park; Masahiro Tamoi; Hironori Inoue; Katsuhiko Sakamoto; Ryota Saito; Chikahiro Miyake

Suppressing a chloroplast oxidoreductase decreases carbon utilization during the night, and inhibits plant growth. Lipid-derived reactive carbonyl species (RCS) possess electrophilic moieties and cause oxidative stress by reacting with cellular components. Arabidopsis (Arabidopsis thaliana) has a chloroplast-localized alkenal/one oxidoreductase (AtAOR) for the detoxification of lipid-derived RCS, especially α,β-unsaturated carbonyls. In this study, we aimed to evaluate the physiological importance of AtAOR and analyzed AtAOR (aor) mutants, including a transfer DNA knockout, aor (T-DNA), and RNA interference knockdown, aor (RNAi), lines. We found that both aor mutants showed smaller plant sizes than wild-type plants when they were grown under day/night cycle conditions. To elucidate the cause of the aor mutant phenotype, we analyzed the photosynthetic rate and the respiration rate by gas-exchange analysis. Subsequently, we found that both wild-type and aor (RNAi) plants showed similar CO2 assimilation rates; however, the respiration rate was lower in aor (RNAi) than in wild-type plants. Furthermore, we revealed that phosphoenolpyruvate carboxylase activity decreased and starch degradation during the night was suppressed in aor (RNAi). In contrast, the phenotype of aor (RNAi) was rescued when aor (RNAi) plants were grown under constant light conditions. These results indicate that the smaller plant sizes observed in aor mutants grown under day/night cycle conditions were attributable to the decrease in carbon utilization during the night. Here, we propose that the detoxification of lipid-derived RCS by AtAOR in chloroplasts contributes to the protection of dark respiration and supports plant growth during the night.

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