Updated on 2024/03/28

写真a

 
HAYASHI Yuki
 
Organization
Graduate School of Science Assistant Professor
Graduate School
Graduate School of Science
Undergraduate School
School of Science Department of Biological Science
Title
Assistant Professor

Degree 1

  1. 博士(理学) ( 2014.3   名古屋大学 ) 

 

Papers 11

  1. Phosphorylation of plasma membrane H(+)-ATPase Thr881 participates in light-induced stomatal opening.

    Hayashi Y, Fukatsu K, Takahashi K, Kinoshita SN, Kato K, Sakakibara T, Kuwata K, Kinoshita T

    Nature communications   Vol. 15 ( 1 ) page: 1194   2024.2

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    Plasma membrane (PM) H+-ATPase is crucial for light-induced stomatal opening and phosphorylation of a penultimate residue, Thr948 (pen-Thr, numbering according to Arabidopsis AHA1) is required for enzyme activation. In this study, a comprehensive phosphoproteomic analysis using guard cell protoplasts from Vicia faba shows that both red and blue light increase the phosphorylation of Thr881, of PM H+-ATPase. Light-induced stomatal opening and the blue light-induced increase in stomatal conductance are reduced in transgenic Arabidopsis plants expressing mutant AHA1-T881A in aha1–9, whereas the blue light-induced phosphorylation of pen-Thr is unaffected. Auxin and photosynthetically active radiation induce the phosphorylation of both Thr881 and pen-Thr in etiolated seedlings and leaves, respectively. The dephosphorylation of phosphorylated Thr881 and pen-Thr are mediated by type 2 C protein phosphatase clade D isoforms. Taken together, Thr881 phosphorylation, in addition of the pen-Thr phosphorylation, are important for PM H+-ATPase function during physiological responses, such as light-induced stomatal opening in Arabidopsis thaliana.

    DOI: 10.1038/s41467-024-45248-5

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  2. Tryptophan synthase ss subunit 1 affects stomatal phenotypes in <i>Arabidopsis thaliana</i>

    Soda, MN; Hayashi, Y; Takahashi, K; Kinoshita, T

    FRONTIERS IN PLANT SCIENCE   Vol. 13   page: 1011360   2022.11

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    Language:English   Publisher:Frontiers in Plant Science  

    Stomata open in response to several environmental stimuli, such as light and low CO2. Plasma membrane (PM) H+-ATPase in guard cells plays a pivotal role for light-induced stomatal opening. In contrast, stomata close in response to the dark or plant hormone abscisic acid (ABA). However, molecular mechanisms of stomatal movements remain unclear. To elucidate the molecular mechanism of stomatal movements, we performed a genetic screen based on stomatal aperture-dependent weight decrease of detached leaves from EMS-treated Arabidopsis thaliana and isolated a rapid transpiration in detached leaves 2 (rtl2). The rtl2 mutant showed constitutive open-stomata phenotype with lower leaf temperature. ABA had no effect on stomatal aperture in rtl2. The rtl2 mutant also showed increased stomatal density, severe dwarf phenotype with pale green leaves and dark veins. Map-based analysis of the RTL2 locus revealed that the rtl2 mutant possesses a single nucleotide substitution, which induces amino acid substitution Gly162 to Glu in the tryptophan synthase ß subunit 1 (TSB1). The TSB1 encodes an enzyme in tryptophan (Trp) biosynthetic pathway. Amount of TSB1 protein was drastically reduced in rtl2 mutant. A different allele of tsb1 mutant (tsb1-1) also showed constitutive open-stomata phenotype with reduced TSB1 protein as in rtl2. Analyses of test-crossed plants of rtl2 and tsb1-1 showed open-stomata and dwarf phenotypes. These results indicate that a responsible gene for rtl2 is TSB1. We further investigated stomatal phenotype in mutants from Trp biosynthetic pathway, such as wei2-1 wei7-1, trp3-1, and tsb2-1. The trp3-1 mutant showed significant wider stomatal aperture as well as tsb1-1. Trp biosynthetic pathway closely relates to auxin biosynthesis. Then, we investigated auxin responsible genes and found that an expression of AUR3 was up in rtl2. In contrast, auxin had no effect on stomatal aperture in Arabidopsis and the phosphorylation status of PM H+-ATPase in guard cell protoplasts from Vicia faba. In addition, auxin antagonist had no effect on stomatal aperture. Interestingly, tsb1-1 grown under hydroponic culture system showed normal stomatal aperture by exogenously application of Trp. These results suggest that open stomata phenotype in tsb1-1 is due to Trp deficiency but not auxin.

    DOI: 10.3389/fpls.2022.1011360

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  3. Type 2C protein phosphatase clade D family members dephosphorylate guard cell plasma membrane H+-ATPase

    Akiyama Mitsumasa, Sugimoto Hodaka, Inoue Shin-ichiro, Takahashi Yohei, Hayashi Maki, Hayashi Yuki, Mizutani Miya, Ogawa Takumi, Kinoshita Daichi, Ando Eigo, Park Meeyeon, Gray William M., Kinoshita Toshinori

    PLANT PHYSIOLOGY   Vol. 188 ( 4 ) page: 2228 - 2240   2022.3

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    Language:Japanese   Publisher:Plant Physiology  

    Plasma membrane (PM) H + -ATPase in guard cells is activated by phosphorylation of the penultimate residue, threonine (Thr), in response to blue and red light, promoting stomatal opening. Previous in vitro biochemical investigation suggested that Mg2 + - and Mn2 + -dependent membrane-localized type 2C protein phosphatase (PP2C)-like activity mediates the dephosphorylation of PM H + -ATPase in guard cells. PP2C clade D (PP2C.D) was later demonstrated to be involved in PM H + -ATPase dephosphorylation during auxin-induced cell expansion in Arabidopsis (Arabidopsis thaliana). However, it is unclear whether PP2C.D phosphatases are involved in PM H + -ATPase dephosphorylation in guard cells. Transient expression experiments using Arabidopsis mesophyll cell protoplasts revealed that all PP2C.D isoforms dephosphorylate the endogenous PM H + -ATPase. We further analyzed PP2C.D6/8/9, which display higher expression levels than other isoforms in guard cells, observing that pp2c.d6, pp2c.d8, and pp2c.d9 single mutants showed similar light-induced stomatal opening and phosphorylation status of PM H + -ATPase in guard cells as Col-0. In contrast, the pp2c.d6/9 double mutant displayed wider stomatal apertures and greater PM H + -ATPase phosphorylation in response to blue light, but delayed dephosphorylation of PM H + -ATPase in guard cells; the pp2c.d6/8/9 triple mutant showed similar phenotypes to those of the pp2c.d6/9 double mutant. Taken together, these results indicate that PP2C.D6 and PP2C.D9 redundantly mediate PM H + -ATPase dephosphorylation in guard cells. Curiously, unlike auxin-induced cell expansion in seedlings, auxin had no effect on the phosphorylation status of PM H + -ATPase in guard cells.

    DOI: 10.1093/plphys/kiab571

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  4. Mechanosensory trichome cells evoke a mechanical stimuli-induced immune response in <i>Arabidopsis thaliana</i>

    Matsumura, M; Nomoto, M; Itaya, T; Aratani, Y; Iwamoto, M; Matsuura, T; Hayashi, Y; Mori, T; Skelly, MJ; Yamamoto, YY; Kinoshita, T; Mori, IC; Suzuki, T; Betsuyaku, S; Spoel, SH; Toyota, M; Tada, Y

    NATURE COMMUNICATIONS   Vol. 13 ( 1 ) page: 1216   2022.3

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    Language:English   Publisher:Nature Communications  

    Perception of pathogen-derived ligands by corresponding host receptors is a pivotal strategy in eukaryotic innate immunity. In plants, this is complemented by circadian anticipation of infection timing, promoting basal resistance even in the absence of pathogen threat. Here, we report that trichomes, hair-like structures on the epidermis, directly sense external mechanical forces, including raindrops, to anticipate pathogen infections in Arabidopsis thaliana. Exposure of leaf surfaces to mechanical stimuli initiates the concentric propagation of intercellular calcium waves away from trichomes to induce defence-related genes. Propagating calcium waves enable effective immunity against pathogenic microbes through the CALMODULIN-BINDING TRANSCRIPTION ACTIVATOR 3 (CAMTA3) and mitogen-activated protein kinases. We propose an early layer of plant immunity in which trichomes function as mechanosensory cells that detect potential risks.

    DOI: 10.1038/s41467-022-28813-8

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  5. Identification of Abscisic Acid-Dependent Phosphorylated Basic Helix-Loop-Helix Transcription Factors in Guard Cells of Vicia faba by Mass Spectrometry

    Hayashi Yuki, Takahashi Yohei, Fukatsu Kohei, Tada Yasuomi, Takahashi Koji, Kuwata Keiko, Suzuki Takamasa, Kinoshita Toshinori

    FRONTIERS IN PLANT SCIENCE   Vol. 12   page: 735271   2021.12

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    Language:Japanese   Publisher:Frontiers in Plant Science  

    An unknown 61 kDa protein is phosphorylated by abscisic acid (ABA)-activated protein kinase in response to ABA and binds to 14-3-3 protein in a phosphorylation-dependent manner in guard-cell protoplasts (GCPs) from Vicia faba. Subsequently, ABA-dependent phosphorylated proteins were identified as basic helix–loop–helix transcription factors, named ABA-responsive kinase substrates (AKSs) in GCPs from Arabidopsis thaliana. However, whether the 61 kDa protein in Vicia GCPs is an AKS is unclear. We performed immunoprecipitation of ABA-treated Vicia GCPs using anti-14-3-3 protein antibodies and identified several AKS isoforms in V. faba (VfAKSs) by mass spectrometry. The 61 kDa protein was identified as VfAKS1. Phosphoproteomic analysis revealed that VfAKSs are phosphorylated at Ser residues, which are important for 14-3-3 protein binding and monomerisation, in response to ABA in GCPs. Orthologs of AtABCG40, an ABA importer in guard cells, and CHC1, a clathrin heavy chain and a regulator of stomatal movement, also co-immunoprecipitated with 14-3-3 protein from guard cells.

    DOI: 10.3389/fpls.2021.735271

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  6. Overexpression of Plasma Membrane H+-ATPase in Guard Cells Enhances Light-Induced Stomatal Opening, Photosynthesis, and Plant Growth in Hybrid Aspen

    Toh Shigeo, Takata Naoki, Ando Eigo, Toda Yosuke, Wang Yin, Hayashi Yuki, Mitsuda Nobutaka, Nagano Soichiro, Taniguchi Toru, Kinoshita Toshinori

    FRONTIERS IN PLANT SCIENCE   Vol. 12   page: 766037   2021.11

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    Language:Japanese   Publisher:Frontiers in Plant Science  

    Stomata in the plant epidermis open in response to light and regulate CO2 uptake for photosynthesis and transpiration for uptake of water and nutrients from roots. Light-induced stomatal opening is mediated by activation of the plasma membrane (PM) H+-ATPase in guard cells. Overexpression of PM H+-ATPase in guard cells promotes light-induced stomatal opening, enhancing photosynthesis and growth in Arabidopsis thaliana. In this study, transgenic hybrid aspens overexpressing Arabidopsis PM H+-ATPase (AHA2) in guard cells under the strong guard cell promoter Arabidopsis GC1 (AtGC1) showed enhanced light-induced stomatal opening, photosynthesis, and growth. First, we confirmed that AtGC1 induces GUS expression specifically in guard cells in hybrid aspens. Thus, we produced AtGC1::AHA2 transgenic hybrid aspens and confirmed expression of AHA2 in AtGC1::AHA2 transgenic plants. In addition, AtGC1::AHA2 transgenic plants showed a higher PM H+-ATPase protein level in guard cells. Analysis using a gas exchange system revealed that transpiration and the photosynthetic rate were significantly increased in AtGC1::AHA2 transgenic aspen plants. AtGC1::AHA2 transgenic plants showed a>20% higher stem elongation rate than the wild type (WT). Therefore, overexpression of PM H+-ATPase in guard cells promotes the growth of perennial woody plants.

    DOI: 10.3389/fpls.2021.766037

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  7. Molecular basis of plasma membrane H+-ATPase function and potential application in the agricultural production

    Ding Ming, Zhang Maoxing, Zeng Houqing, Hayashi Yuki, Zhu Yiyong, Kinoshita Toshinori

    PLANT PHYSIOLOGY AND BIOCHEMISTRY   Vol. 168   page: 10 - 16   2021.11

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    Language:Japanese   Publisher:Plant Physiology and Biochemistry  

    Increase of crop yield is always the desired goal, manipulation of genes in relation to plant growth is a shortcut to promote crop yield. The plasma membrane (PM) H+-ATPase is the plant master enzyme; the energy yielded by ATP hydrolysis pumps H+ out of cells, establishes the membrane potential, maintains pH homeostasis and provides the proton-motive force required for transmembrane transport of many materials. PM H+-ATPase is involved in root nutrient uptake, epidermal stomatal opening, phloem sucrose loading and unloading, and hypocotyl cell elongation. In this review, we summarize the recent progresses in roles of PM H+-ATPase in nutrient uptake and light-induced stomatal opening and discuss the pivotal role of PM H+-ATPase in crop yield improvement and its potential application in agricultural production by modulating the expression of PM H+-ATPase in crops.

    DOI: 10.1016/j.plaphy.2021.09.036

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  8. Plasma membrane H+-ATPase overexpression increases rice yield via simultaneous enhancement of nutrient uptake and photosynthesis

    Zhang Maoxing, Wang Yin, Chen Xi, Xu Feiyun, Ding Ming, Ye Wenxiu, Kawai Yuya, Toda Yosuke, Hayashi Yuki, Suzuki Takamasa, Zeng Houqing, Xiao Liang, Xiao Xin, Xu Jin, Guo Shiwei, Yan Feng, Shen Qirong, Xu Guohua, Kinoshita Toshinori, Zhu Yiyong

    NATURE COMMUNICATIONS   Vol. 12 ( 1 ) page: 735   2021.2

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    Language:Japanese   Publisher:Nature Communications  

    Nitrogen (N) and carbon (C) are essential elements for plant growth and crop yield. Thus, improved N and C utilisation contributes to agricultural productivity and reduces the need for fertilisation. In the present study, we find that overexpression of a single rice gene, Oryza sativa plasma membrane (PM) H+-ATPase 1 (OSA1), facilitates ammonium absorption and assimilation in roots and enhanced light-induced stomatal opening with higher photosynthesis rate in leaves. As a result, OSA1 overexpression in rice plants causes a 33% increase in grain yield and a 46% increase in N use efficiency overall. As PM H+-ATPase is highly conserved in plants, these findings indicate that the manipulation of PM H+-ATPase could cooperatively improve N and C utilisation, potentially providing a vital tool for food security and sustainable agriculture.

    DOI: 10.1038/s41467-021-20964-4

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  9. Identification of Genes Preferentially Expressed in Stomatal Guard Cells of <i>Arabidopsis thaliana</i> and Involvement of the Aluminum-Activated Malate Transporter 6 Vacuolar Malate Channel in Stomatal Opening.

    Ye W, Koya S, Hayashi Y, Jiang H, Oishi T, Kato K, Fukatsu K, Kinoshita T

    Frontiers in plant science   Vol. 12   page: 744991   2021

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    Stomatal guard cells (GCs) are highly specialized cells that respond to various stimuli, such as blue light (BL) and abscisic acid, for the regulation of stomatal aperture. Many signaling components that are involved in the stomatal movement are preferentially expressed in GCs. In this study, we identified four new such genes in addition to an aluminum-activated malate transporter, ALMT6, and GDSL lipase, Occlusion of Stomatal Pore 1 (OSP1), based on the expression analysis using public resources, reverse transcription PCR, and promoter-driven β-glucuronidase assays. Some null mutants of GC-specific genes evidenced altered stomatal movement. We further investigated the role played by ALMT6, a vacuolar malate channel, in stomatal opening. Epidermal strips from an ALMT6-null mutant exhibited defective stomatal opening induced by BL and fusicoccin, a strong plasma membrane H+-ATPase activator. The deficiency was enhanced when the assay buffer [Cl–] was low, suggesting that malate and/or Cl– facilitate efficient opening. The results indicate that the GC-specific genes are frequently involved in stomatal movement. Further detailed analyses of the hitherto uncharacterized GC-specific genes will provide new insights into stomatal regulation.

    DOI: 10.3389/fpls.2021.744991

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  10. Regulation of stomatal opening and histone modification by photoperiod in Arabidopsis thaliana

    Aoki Saya, Toh Shigeo, Nakamichi Norihito, Hayashi Yuki, Wang Yin, Suzuki Takamasa, Tsuji Hiroyuki, Kinoshita Toshinori

    SCIENTIFIC REPORTS   Vol. 9 ( 1 ) page: 10054   2019.7

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    Language:Japanese   Publisher:Scientific Reports  

    Stomatal movements are regulated by many environmental signals, such as light, CO2, temperature, humidity, and drought. Recently, we showed that photoperiodic flowering components have positive effects on light-induced stomatal opening in Arabidopsis thaliana. In this study, we determined that light-induced stomatal opening and increased stomatal conductance were larger in plants grown under long-day (LD) conditions than in those grown under short-day (SD) conditions. Gene expression analyses using purified guard cell protoplasts revealed that FT and SOC1 expression levels were significantly increased under LD conditions. Interestingly, the enhancement of light-induced stomatal opening and increased SOC1 expression in guard cells due to LD conditions persisted for at least 1 week after plants were transferred to SD conditions. We then investigated histone modification using chromatin immunoprecipitation–PCR, and observed increased trimethylation of lysine 4 on histone 3 (H3K4) around SOC1. We also found that LD-dependent enhancement of light-induced stomatal opening and H3K4 trimethylation in SOC1 were suppressed in the ft-2 mutant. These results indicate that photoperiod is an important environmental cue regulating stomatal opening, and that LD conditions enhance light-induced stomatal opening and epigenetic modification (H3K4 trimethylation) around SOC1, a positive regulator of stomatal opening, in an FT-dependent manner. Thus, this study provides novel insights into stomatal responses to photoperiod.

    DOI: 10.1038/s41598-019-46440-0

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  11. Control of seed dormancy and germination by DOG1-AHG1 PP2C phosphatase complex via binding to heme

    Nishimura Noriyuki, Tsuchiya Wataru, Moresco James J., Hayashi Yuki, Satoh Kouji, Kaiwa Nahomi, Irisa Tomoko, Kinoshita Toshinori, Schroeder Julian I., Yates John R. III, Hirayama Takashi, Yamazaki Toshimasa

    NATURE COMMUNICATIONS   Vol. 9 ( 1 ) page: 2132   2018.6

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    Abscisic acid (ABA) regulates abiotic stress and developmental responses including regulation of seed dormancy to prevent seeds from germinating under unfavorable environmental conditions. ABA HYPERSENSITIVE GERMINATION1 (AHG1) encoding a type 2C protein phosphatase (PP2C) is a central negative regulator of ABA response in germination; however, the molecular function and regulation of AHG1 remain elusive. Here we report that AHG1 interacts with DELAY OF GERMINATION1 (DOG1), which is a pivotal positive regulator in seed dormancy. DOG1 acts upstream of AHG1 and impairs the PP2C activity of AHG1 in vitro. Furthermore, DOG1 has the ability to bind heme. Binding of DOG1 to AHG1 and heme are independent processes, but both are essential for DOG1 function in vivo. Our study demonstrates that AHG1 and DOG1 constitute an important regulatory system for seed dormancy and germination by integrating multiple environmental signals, in parallel with the PYL/RCAR ABA receptor-mediated regulatory system.

    DOI: 10.1038/s41467-018-04437-9

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Books 2

  1. 植物の通気口「気孔」ー環境に応答する気孔の制御

    林優紀、木下俊則( Role: Joint author)

    2020.9 

  2. 植物の巧みな環境応答 ー植物の情報処理研究の最前線

    木下俊則、林優紀( Role: Joint author)

    2020.9