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

     More details

    Language:English   Publisher:Nature Communications  

    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

    Scopus

    PubMed

  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

     More details

    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

    Web of Science

    Scopus

    PubMed

  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

     More details

    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

    Web of Science

    Scopus

    PubMed

  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

     More details

    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

    Web of Science

    Scopus

    PubMed

  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

     More details

    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

    Web of Science

    Scopus

    PubMed

▼display all

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