Performance information

• Loss-of-functional mutation in ANGUSTIFOLIA3 causes leucine hypersensitivity and hypoxia response during Arabidopsis thaliana seedling growth　　　　　　　　　　　　　　　
  Kensuke Kawade; Mamoru Nozaki; Gorou Horiguchi; Tomoko Mori; Katsushi Yamaguchi; Mami Okamoto; Hiromitsu Tabeta; Shuji Shigenobu; Masami Yokota Hirai; Hirokazu Tsukaya
  Metabolomics, Mar. 2025, [Reviewed], [Lead, Corresponding]
  Scientific journal
  DOI：https://doi.org/10.1007/s11306-025-02249-9
  DOI ID：10.1007/s11306-025-02249-9, ORCID：181412372
• Functional conservation and divergence of Arabidopsis VENOSA4 and human SAMHD1 in DNA repair　　　　　　　　　　　　　　　
  Raquel Sarmiento-Mañús; Sara Fontcuberta-Cervera; Kensuke Kawade; Akira Oikawa; Hirokazu Tsukaya; Víctor Quesada; José Luis Micol; María Rosa Ponce
  Heliyon, May 2024, [Reviewed]
  DOI：https://doi.org/10.1101/2024.05.27.596045
  DOI ID：10.1101/2024.05.27.596045, ORCID：160687091
• Control of root nodule formation ensures sufficient shoot water availability in Lotus japonicus
  Kensuke Kawade; Daisuke Sugiura; Akira Oikawa; Masayoshi Kawaguchi
  Plant Physiology, Mar. 2024, [Reviewed], [Lead, Corresponding]
  Abstract
  
  Leguminous plants provide carbon to symbiotic rhizobia in root nodules to fuel the energy-consuming process of nitrogen fixation. The carbon investment pattern from the acquired sources is crucial for shaping the growth regime of the host plants. The autoregulation of nodulation (AON) signaling pathway tightly regulates the number of nodules that form. AON disruption leads to excessive nodule formation and stunted shoot growth. However, the physiological role of AON in adjusting the carbon investment pattern is unknown. Here, we show that AON plays an important role in sustaining shoot water availability, which is essential for promoting carbon investment in shoot growth in Lotus japonicus. We found that AON-defective mutants exhibit substantial accumulation of nonstructural carbohydrates, such as sucrose. Consistent with this metabolic signature, resilience against water-deficit stress was enhanced in the shoots of the AON-defective mutants. Furthermore, the water uptake ability was attenuated in the AON-defective mutants, likely due to the increased ratio of nodulation zone, which is covered with hydrophobic surfaces, on the roots. These results increase our physiological understanding of legume–rhizobia symbiosis by revealing a trade-off between root nodule formation and shoot water availability.
  Oxford University Press (OUP), Scientific journal
  DOI：https://doi.org/10.1093/plphys/kiae126
  DOI ID：10.1093/plphys/kiae126, ISSN：0032-0889, eISSN：1532-2548
• IMA peptides regulate root nodulation and nitrogen homeostasis by providing iron according to internal nitrogen status
  Momoyo Ito; Yuri Tajima; Mari Ogawa-Ohnishi; Hanna Nishida; Shohei Nosaki; Momona Noda; Naoyuki Sotta; Kensuke Kawade; Takehiro Kamiya; Toru Fujiwara; Yoshikatsu Matsubayashi; Takuya Suzaki
  Nature Communications, Volume：15, Number：1, Jan. 2024, [Reviewed]
  Abstract
  
  Legumes control root nodule symbiosis (RNS) in response to environmental nitrogen availability. Despite the recent understanding of the molecular basis of external nitrate-mediated control of RNS, it remains mostly elusive how plants regulate physiological processes depending on internal nitrogen status. In addition, iron (Fe) acts as an essential element that enables symbiotic nitrogen fixation; however, the mechanism of Fe accumulation in nodules is poorly understood. Here, we focus on the transcriptome in response to internal nitrogen status during RNS in Lotus japonicus and identify that IRON MAN (IMA) peptide genes are expressed during symbiotic nitrogen fixation. We show that LjIMA1 and LjIMA2 expressed in the shoot and root play systemic and local roles in concentrating internal Fe to the nodule. Furthermore, IMA peptides have conserved roles in regulating nitrogen homeostasis by adjusting nitrogen-Fe balance in L. japonicus and Arabidopsis thaliana. These findings indicate that IMA-mediated Fe provision plays an essential role in regulating nitrogen-related physiological processes.
  Springer Science and Business Media LLC, Scientific journal
  DOI：https://doi.org/10.1038/s41467-024-44865-4
  DOI ID：10.1038/s41467-024-44865-4, eISSN：2041-1723
• The Roles of Functional Amino Acids in Plant Growth and Development
  Kensuke Kawade; Hiromitsu Tabeta; Ali Ferjani; Masami Yokota Hirai
  Plant and Cell Physiology, Jul. 2023, [Reviewed], [Lead, Corresponding]
  Abstract
  
  Plants incorporate acquired carbon and nitrogen into amino acid metabolism, whereby the building blocks of proteins and the precursors of various metabolites are produced. This fundamental demand requires tight amino acid metabolism to sustain physiological homeostasis. There is increasing evidence that amino acid metabolism undergoes plastic alteration to orchestrate specific growth and developmental events. Consequently, there has been a gradual exploration of the interface at which amino acid metabolism and plant morphogenesis are mutually affected. This research progress offers an opportunity to explore amino acid metabolism, with the goal to understand how it can be modulated to serve special cellular needs and regulate specific growth and developmental pathways. Continuous improvements in the sensitivity and coverage of metabolomics technology, along with the development of chemoinformatics, have allowed the investigation of these research questions. In this review, we summarize the roles of threonine, serine, arginine and γ-aminobutyric acid as representative examples of amino acids relevant to specific developmental processes in plants (‘functional amino acids’). Our objective is to expand perspectives regarding amino acid metabolism beyond the conventional view that it is merely life-supporting machinery.
  Oxford University Press (OUP), Scientific journal
  DOI：https://doi.org/10.1093/pcp/pcad071
  DOI ID：10.1093/pcp/pcad071, ISSN：0032-0781, eISSN：1471-9053
• Leaf-size control beyond transcription factors: Compensatory mechanisms
  Hiromitsu Tabeta; Shizuka Gunji; Kensuke Kawade; Ali Ferjani
  Frontiers in Plant Science, Volume：13, Jan. 2023, [Reviewed]
  Plant leaves display abundant morphological richness yet grow to characteristic sizes and shapes. Beginning with a small number of undifferentiated founder cells, leaves evolve via a complex interplay of regulatory factors that ultimately influence cell proliferation and subsequent post-mitotic cell enlargement. During their development, a sequence of key events that shape leaves is both robustly executed spatiotemporally following a genomic molecular network and flexibly tuned by a variety of environmental stimuli. Decades of work on Arabidopsis thaliana have revisited the compensatory phenomena that might reflect a general and primary size-regulatory mechanism in leaves. This review focuses on key molecular and cellular events behind the organ-wide scale regulation of compensatory mechanisms. Lastly, emerging novel mechanisms of metabolic and hormonal regulation are discussed, based on recent advances in the field that have provided insights into, among other phenomena, leaf-size regulation.
  Frontiers Media SA, Scientific journal
  DOI：https://doi.org/10.3389/fpls.2022.1024945
  DOI ID：10.3389/fpls.2022.1024945, eISSN：1664-462X
• Tissue-targeted inorganic pyrophosphate hydrolysis in a fugu5 mutant reveals that excess inorganic pyrophosphate triggers developmental defects in a cell-autonomous manner
  Shizuka Gunji; Kensuke Kawade; Hiromitsu Tabeta; Gorou Horiguchi; Akira Oikawa; Mariko Asaoka; Masami Yokota Hirai; Hirokazu Tsukaya; Ali Ferjani
  Frontiers in Plant Science, Volume：13, Aug. 2022, [Reviewed]
  Excess PPi triggers developmental defects in a cell-autonomous manner. The level of inorganic pyrophosphate (PPi) must be tightly regulated in all kingdoms for the proper execution of cellular functions. In plants, the vacuolar proton pyrophosphatase (H⁺-PPase) has a pivotal role in PPi homeostasis. We previously demonstrated that the excess cytosolic PPi in the H⁺-PPase loss-of-function fugu5 mutant inhibits gluconeogenesis from seed storage lipids, arrests cell division in cotyledonary palisade tissue, and triggers a compensated cell enlargement (CCE). Moreover, PPi alters pavement cell (PC) shape, stomatal patterning, and functioning, supporting specific yet broad inhibitory effects of PPi on leaf morphogenesis. Whereas these developmental defects were totally rescued by the expression of the yeast soluble pyrophosphatase IPP1, sucrose supply alone canceled CCE in the palisade tissue but not the epidermal developmental defects. Hence, we postulated that the latter are likely triggered by excess PPi rather than a sucrose deficit. To formally test this hypothesis, we adopted a spatiotemporal approach by constructing and analyzing fugu5-1 PDF1_pro::IPP1, fugu5-1 CLV1_pro::IPP1, and fugu5-1 ICL_pro::IPP1, whereby PPi was removed specifically from the epidermis, palisade tissue cells, or during the 4 days following seed imbibition, respectively. It is important to note that whereas PC defects in fugu5-1 PDF1_pro::IPP1 were completely recovered, those in fugu5-1 CLV1_pro::IPP1 were not. In addition, phenotypic analyses of fugu5-1 ICL_pro::IPP1 lines demonstrated that the immediate removal of PPi after seed imbibition markedly improved overall plant growth, abolished CCE, but only partially restored the epidermal developmental defects. Next, the impact of spatial and temporal removal of PPi was investigated by capillary electrophoresis time-of-flight mass spectrometry (CE-TOF MS). Our analysis revealed that the metabolic profiles are differentially affected among all the above transgenic lines, and consistent with an axial role of central metabolism of gluconeogenesis in CCE. Taken together, this study provides a conceptual framework to unveil metabolic fluctuations within leaf tissues with high spatio–temporal resolution. Finally, our findings suggest that excess PPi exerts its inhibitory effect in planta in the early stages of seedling establishment in a tissue- and cell-autonomous manner.
  Frontiers Media SA, Scientific journal
  DOI：https://doi.org/10.3389/fpls.2022.945225
  DOI ID：10.3389/fpls.2022.945225, eISSN：1664-462X
• Metabolic Control of Gametophore Shoot Formation through Arginine in the Moss Physcomitrium patens　　　　　　　　　　　　　　　
  Kensuke Kawade; Gorou Horiguchi; Yuu Hirose; Akira Oikawa; Masami Yokota Hirai; Kazuki Saito; Tomomichi Fujita; Hirokazu Tsukaya
  Cell Reports, Volume：32, Number：10, First page：108127, Last page：108127, Sep. 2020, [Reviewed], [Lead, Corresponding], ［International magazine］
  Shoot formation is accompanied by active cell proliferation and expansion, requiring that metabolic state adapts to developmental control. Despite the importance of such metabolic reprogramming, it remains unclear how development and metabolism are integrated. Here, we show that disruption of ANGUSTIFOLIA3 orthologs (PpAN3s) compromises gametophore shoot formation in the moss Physcomitrium patens due to defective cell proliferation and expansion. Trans-omics analysis reveals that the downstream activity of PpAN3 is linked to arginine metabolism. Elevating arginine level by chemical treatment leads to stunted gametophores and causes Ppan3 mutant-like transcriptional changes in the wild-type plant. Furthermore, ectopic expression of AtAN3 from Arabidopsis thaliana ameliorates the defective arginine metabolism and promotes gametophore formation in Ppan3 mutants. Together, these findings indicate that arginine metabolism is a key pathway associated with gametophore formation and provide evolutionary insights into the establishment of the shoot system in land plants through the integration of developmental and metabolic processes.
  Elsevier {BV}, English, Scientific journal
  DOI：https://doi.org/10.1016/j.celrep.2020.108127
  DOI ID：10.1016/j.celrep.2020.108127, ORCID：80025818, PubMed ID：32905770
• an3-Mediated Compensation Is Dependent on a Cell-Autonomous Mechanism in Leaf Epidermal Tissue　　　　　　　　　　　　　　　
  Mamoru Nozaki; Kensuke Kawade; Gorou Horiguchi; Hirokazu Tsukaya
  Plant and Cell Physiology, Volume：61, Number：6, First page：1181, Last page：1190, Jun. 2020, [Reviewed], [Corresponding], ［Domestic magazine］
  Leaves are formed by coordinated growth of tissue layers driven by cell proliferation and expansion. Compensation, in which a defect in cell proliferation induces compensated cell enlargement (CCE), plays an important role in cell-size determination during leaf development. We previously reported that CCE triggered by the an3 mutation is observed in epidermal and subepidermal layers in Arabidopsis thaliana (Arabidopsis) leaves. Interestingly, CCE is induced in a non-cell autonomous manner between subepidermal cells. However, whether CCE in the subepidermis affects cell size in the adjacent epidermis is still unclear. We induced layer-specific expression of AN3 in an3 leaves and found that CCE in the subepidermis had little impact on cell-size determination in the epidermis, and vice versa, suggesting that CCE is induced in a tissue-autonomous manner. Examination of the epidermis in an3 leaves having AN3-positive and -negative sectors generated by Cre/loxP revealed that, in contrast to the subepidermis, CCE occurred exclusively in AN3-negative epidermal cells, indicating a cell autonomous action of an3-mediated compensation in the epidermis. These results clarified that the epidermal and subepidermal tissue layers have different cell autonomies in CCE. In addition, quantification of cell-expansion kinetics in epidermal and subepidermal tissues of the an3 showed that the tissues exhibited a similar temporal profile to reach a peak cell-expansion rate as compared to wild type. This might be one feature representing that the two tissue layers retain their growth coordination even in the presence of CCE.
  Oxford University Press ({OUP}), English, Scientific journal
  DOI：https://doi.org/10.1093/pcp/pcaa048
  DOI ID：10.1093/pcp/pcaa048, ORCID：75676187, PubMed ID：32321167
• Quantitative Imaging Reveals Distinct Contributions of SnRK2 and ABI3 in Plasmodesmatal Permeability in Physcomitrella patens
  Takumi Tomoi; Kensuke Kawade; Munenori Kitagawa; Yoichi Sakata; Hirokazu Tsukaya; Tomomichi Fujita
  Plant and Cell Physiology, Volume：61, Number：5, First page：942, Last page：956, May 2020, [Reviewed], [Corresponding]
  
  Cell-to-cell communication is tightly regulated in response to environmental stimuli in plants. We previously used a photoconvertible fluorescent protein Dendra2 as a model reporter to study this process. This experiment revealed that macromolecular trafficking between protonemal cells in Physcomitrella patens is suppressed in response to abscisic acid (ABA). However, it remains unknown which ABA signaling components contribute to this suppression and how. Here, we show that ABA signaling components SUCROSE NON-FERMENTING 1-RELATED PROTEIN KINASE 2 (PpSnRK2) and ABA INSENSITIVE 3 (PpABI3) play roles as an essential and promotive factor, respectively, in regulating ABA-induced suppression of Dendra2 diffusion between cells (ASD). Our quantitative imaging analysis revealed that disruption of PpSnRK2 resulted in defective ASD onset itself, whereas disruption of PpABI3 caused an 81-min delay in the initiation of ASD. Live-cell imaging of callose deposition using aniline blue staining showed that, despite this onset delay, callose deposition on cross walls remained constant in the PpABI3 disruptant, suggesting that PpABI3 facilitates ASD in a callose-independent manner. Given that ABA is an important phytohormone to cope with abiotic stresses, we further explored cellular physiological responses. We found that the acquisition of salt stress tolerance is promoted by PpABI3 in a quantitative manner similar to ASD. Our results suggest that PpABI3-mediated ABA signaling may effectively coordinate cell-to-cell communication during the acquisition of salt stress tolerance. This study will accelerate the quantitative study for ABA signaling mechanism and function in response to various abiotic stresses.
  Oxford University Press ({OUP}), Scientific journal
  DOI：https://doi.org/10.1093/pcp/pcaa021
  DOI ID：10.1093/pcp/pcaa021, ISSN：0032-0781, eISSN：1471-9053, ORCID：71631777
• Pyrophosphate inhibits gluconeogenesis by restricting UDP-glucose formation in vivo　　　　　　　　　　　　　　　
  Ali Ferjani; Kensuke Kawade; Mariko Asaoka; Akira Oikawa; Takashi Okada; Atsushi Mochizuki; Masayoshi Maeshima; Masami Yokota Hirai; Kazuki Saito; Hirokazu Tsukaya
  Scientific Reports, Volume：8, Number：1, First page：14696, Last page：14696, Dec. 2018, [Reviewed], ［International magazine］
  Pyrophosphate (PPi) is produced by anabolic reactions and serves as an energy donor in the cytosol of plant cells; however, its accumulation to toxic levels disrupts several common biosynthetic pathways and is lethal. Before acquiring photosynthetic capacity, young seedlings must endure a short but critical heterotrophic period, during which they are nourished solely by sugar produced from seed reserves by the anabolic process of gluconeogenesis. Previously, we reported that excess PPi in H+-PPase-knockout fugu5 mutants of Arabidopsis thaliana severely compromised gluconeogenesis. However, the precise metabolic target of PPi inhibition in vivo remained elusive. Here, CE-TOF MS analyses of major metabolites characteristic of gluconeogenesis from seed lipids showed that the Glc6P;Fru6P level significantly increased and that Glc1P level was consistently somewhat higher in fugu5 compared to wild type. In contrast, the UDP-Glc level decreased significantly in the mutants. Importantly, specific removal of PPi in fugu5, and thus in AVP1pro:IPP1 transgenic lines, restored the Glc1P and the Glc6P;Fru6P levels, increased the UDP-Glc level ~2.0-fold, and subsequently increased sucrose synthesis. Given the reversible nature of the Glc1P/UDP-Glc reaction, our results indicate that UGP-Glc pyrophosphorylase is the major target when excess PPi exerts inhibitory effects in vivo. To validate our findings, we analyzed metabolite responses using a mathematical theory called structural sensitivity analysis (SSA), in which the responses of concentrations in reaction systems to perturbations in enzyme activity are determined from the structure of the network alone. A comparison of our experimental data with the results of pure structural theory predicted the existence of unknown reactions as the necessary condition for the above metabolic profiles, and confirmed the above results. Our data support the notion that H+-PPase plays a pivotal role in cytosolic PPi homeostasis in plant cells. We propose that the combination of metabolomics and SSA is powerful when seeking to identify and predict metabolic targets in living cells.
  Springer Science and Business Media {LLC}, English, Scientific journal
  DOI：https://doi.org/10.1038/s41598-018-32894-1
  DOI ID：10.1038/s41598-018-32894-1, ORCID：48634905, PubMed ID：30279540, PubMed Central ID：PMC6168488
• The cytochrome P450 CYP77A4 is involved in auxin-mediated patterning of the Arabidopsis thaliana embryo　　　　　　　　　　　　　　　
  Kensuke Kawade; Yimeng Li; Hiroyuki Koga; Yuji Sawada; Mami Okamoto; Ayuko Kuwahara; Hirokazu Tsukaya; Masami Yokota Hirai
  Development, Volume：145, First page：dev168369, Sep. 2018, [Reviewed], [Lead, Corresponding]
  Scientific journal
  DOI：https://doi.org/10.1242/dev.168369
  DOI ID：10.1242/dev.168369, ORCID：48237685
• Probing the stochastic property of endoreduplication in cell size determination of Arabidopsis thaliana leaf epidermal tissue　　　　　　　　　　　　　　　
  Kensuke Kawade; Hirokazu Tsukaya
  PLOS ONE, Volume：12, Number：9, First page：e0185050, Sep. 2017, [Reviewed], [Lead, Corresponding]
  Public Library of Science ({PLoS}), Scientific journal
  DOI：https://doi.org/10.1371/journal.pone.0185050
  DOI ID：10.1371/journal.pone.0185050, ORCID：36993988, PubMed ID：28926847
• Spatially Different Tissue-Scale Diffusivity Shapes ANGUSTIFOLIA3 Gradient in Growing Leaves.　　　　　　　　　　　　　　　
  Kensuke Kawade; Hirokazu Tanimoto; Gorou Horiguchi; Hirokazu Tsukaya
  Biophysical Journal, Volume：113, Number：5, First page：1109, Last page：1120, Sep. 2017, [Reviewed], [Lead, Corresponding], ［International magazine］
  The spatial gradient of signaling molecules is pivotal for establishing developmental patterns of multicellular organisms. It has long been proposed that these gradients could arise from the pure diffusion process of signaling molecules between cells, but whether this simplest mechanism establishes the formation of the tissue-scale gradient remains unclear. Plasmodesmata are unique channel structures in plants that connect neighboring cells for molecular transport. In this study, we measured cellular- and tissue-scale kinetics of molecular transport through plasmodesmata in Arabidopsis thaliana developing leaf primordia by fluorescence recovery assays. These trans-scale measurements revealed biophysical properties of diffusive molecular transport through plasmodesmata and revealed that the tissue-scale diffusivity, but not the cellular-scale diffusivity, is spatially different along the leaf proximal-to-distal axis. We found that the gradient in cell size along the developmental axis underlies this spatially different tissue-scale diffusivity. We then asked how this diffusion-based framework functions in establishing a signaling gradient of endogenous molecules. ANGUSTIFOLIA3 (AN3) is a transcriptional co-activator, and as we have shown here, it forms a long-range signaling gradient along the leaf proximal-to-distal axis to determine a cell-proliferation domain. By genetically engineering AN3 mobility, we assessed each contribution of cell-to-cell movement and tissue growth to the distribution of the AN3 gradient. We constructed a diffusion-based theoretical model using these quantitative data to analyze the AN3 gradient formation and demonstrated that it could be achieved solely by the diffusive molecular transport in a growing tissue. Our results indicate that the spatially different tissue-scale diffusivity is a core mechanism for AN3 gradient formation. This provides evidence that the pure diffusion process establishes the formation of the long-range signaling gradient in leaf development.
  English, Scientific journal
  DOI：https://doi.org/10.1016/j.bpj.2017.06.072
  DOI ID：10.1016/j.bpj.2017.06.072, ORCID：36993982, PubMed ID：28877493, PubMed Central ID：PMC5611674
• A novel method for single-grain-based metabolic profiling of Arabidopsis seed　　　　　　　　　　　　　　　
  Yuji Sawada; Hirokazu Tsukaya; Yimeng Li; Muneo Sato; Kensuke Kawade; Masami Yokota Hirai
  Metabolomics, Volume：13, Number：6, May 2017, [Reviewed]
  Introduction In plant metabolomics, metabolite contents are often normalized by sample weight. However, accurate weighing of very small samples, such as individual Arabidopsis thaliana seeds ( approximately 20 mu g), is difficult, which may lead to irreproducible results.Objectives We aimed to establish alternative normalization methods for seed-grain-based comparative metabolomics of A. thaliana.Methods Arabidopsis thaliana seeds were assumed to have a prolate spheroid shape. Using a microscope image of each seed, the lengths of major and minor axes were measured by fitting a projected 2-dimensional shape of each seed as an ellipse. Metabolic profiles of individual diploid or tetraploid A. thaliana seeds were measured by our highly sensitive protocol ("widely targeted metabolomics") that uses liquid chromatography coupled with tandem quadrupole mass spectrometry. Mass spectrometric analysis of 1 mu L of solution extract identified more than 100 metabolites. The data were normalized by various seed-size measures, including seed volume ( single-grain-based analysis). For comparison, metabolites were extracted from 4 mg of diploid and tetraploid A. thaliana seeds and their metabolic profiles were analyzed by normalization of weight (weight-based analysis).Results A small number of metabolites showed statistically significant differences in the single-grain-based analysis compared to weight-based analysis. A total of 17 metabolites showed statistically different accumulation between ploidy types with similar fold changes in both analyses.Conclusion Seed-size measures obtained by microscopic imaging were useful for data normalization. Single-grain-based analysis enables evaluation of metabolism of each seed and elucidates the metabolic profiles of precious bioresources by using small amounts of samples.
  Springer Nature, English, Scientific journal
  DOI：https://doi.org/10.1007/s11306-017-1211-1
  DOI ID：10.1007/s11306-017-1211-1, ISSN：1573-3882, eISSN：1573-3890, ORCID：33950835, Web of Science ID：WOS:000401711400010
• Detection of the Cell Proliferation Zone in Leaves by Using EdU　　　　　　　　　　　　　　　
  Hokuto Nakayama; Kensuke Kawade; Hirokazu Tsukaya; Seisuke Kimura
  BIO-PROTOCOL, Volume：5, Number：18, Sep. 2015, [Reviewed]
  Bio-Protocol, {LLC}, Scientific journal
  DOI：https://doi.org/10.21769/bioprotoc.1600
  DOI ID：10.21769/bioprotoc.1600, ORCID：33950834
• Compensation: a key to clarifying the organ-level regulation of lateral organ size in plants　　　　　　　　　　　　　　　
  T. Hisanaga; K. Kawade; H. Tsukaya
  Journal of Experimental Botany, Volume：66, Number：4, First page：1055, Last page：1063, Jan. 2015, [Reviewed]
  Oxford University Press ({OUP}), Scientific journal
  DOI：https://doi.org/10.1093/jxb/erv028
  DOI ID：10.1093/jxb/erv028, ORCID：33951123
• Mobility of signaling molecules: the key to deciphering plant organogenesis　　　　　　　　　　　　　　　
  Kensuke Kawade; Hirokazu Tanimoto
  Journal of Plant Research, Volume：128, Number：1, First page：17, Last page：25, Jan. 2015, [Reviewed], [Lead, Corresponding]
  Springer Nature, Scientific journal
  DOI：https://doi.org/10.1007/s10265-014-0692-5
  DOI ID：10.1007/s10265-014-0692-5, ORCID：33950983
• Proliferative control of leaf cells through inter-cell-layer AN3 signaling　　　　　　　　　　　　　　　
  Kensuke Kawade
  PLANT MORPHOLOGY, Volume：26, Number：1, First page：59, Last page：63, Apr. 2014, [Reviewed], [Lead, Corresponding]
  The Japanese Society of Plant Morphology, Scientific journal
  DOI：https://doi.org/10.5685/plmorphol.26.59
  DOI ID：10.5685/plmorphol.26.59, ORCID：33951051
• Promotion of chloroplast proliferation upon enhanced post-mitotic cell expansion in leaves　　　　　　　　　　　　　　　
  Kensuke Kawade; Gorou Horiguchi; Naoko Ishikawa; Masami Hirai; Hirokazu Tsukaya
  BMC Plant Biology, Volume：13, Number：1, First page：143, Last page：143, Sep. 2013, [Reviewed], [Lead, Corresponding], ［International magazine］
  BACKGROUND: Leaves are determinate organs; hence, precise control of cell proliferation and post-mitotic cell expansion is essential for their growth. A defect in cell proliferation often triggers enhanced post-mitotic cell expansion in leaves. This phenomenon is referred to as 'compensation'. Several lines of evidence from studies on compensation have shown that cell proliferation and post-mitotic cell expansion are coordinately regulated during leaf development. Therefore, compensation has attracted much attention to the mechanisms for leaf growth. However, our understanding of compensation at the subcellular level remains limited because studies of compensation have focused mainly on cellular-level phenotypes. Proper leaf growth requires quantitative control of subcellular components in association with cellular-level changes. To gain insight into the subcellular aspect of compensation, we investigated the well-known relationship between cell area and chloroplast number per cell in compensation-exhibiting lines, and asked whether chloroplast proliferation is modulated in response to the induction of compensation. RESULTS: We first established a convenient and reliable method for observation of chloroplasts in situ. Using this method, we analyzed Arabidopsis thaliana mutants fugu5 and angustifolia3 (an3), and a transgenic line KIP-RELATED PROTEIN2 overexpressor (KRP2 OE), which are known to exhibit typical features of compensation. We here showed that chloroplast number per cell increased in the subepidermal palisade tissue of these lines. We analyzed tetraploidized wild type, fugu5, an3 and KRP2 OE, and found that cell area itself, but not nuclear ploidy, is a key parameter that determines the activity of chloroplast proliferation. In particular, in the case of an3, we uncovered that promotion of chloroplast proliferation depends on the enhanced post-mitotic cell expansion. The expression levels of chloroplast proliferation-related genes are similar to or lower than that in the wild type during this process. CONCLUSIONS: This study demonstrates that chloroplast proliferation is promoted in compensation-exhibiting lines. This promotion of chloroplast proliferation takes place in response to cell-area increase in post-mitotic phase in an3. The expression of chloroplast proliferation-related genes were not promoted in compensation-exhibiting lines including an3, arguing that an as-yet-unknown mechanism is responsible for modulation of chloroplast proliferation in these lines.
  Springer Nature, English, Scientific journal
  DOI：https://doi.org/10.1186/1471-2229-13-143
  DOI ID：10.1186/1471-2229-13-143, ORCID：33951118, PubMed ID：24074400, PubMed Central ID：PMC3849334
• ANGUSTIFOLIA3 Signaling Coordinates Proliferation between Clonally Distinct Cells in Leaves　　　　　　　　　　　　　　　
  Kensuke Kawade; Gorou Horiguchi; Takeshi Usami; Masami Yokota Hirai; Hirokazu Tsukaya
  Current Biology, Volume：23, Number：9, First page：788, Last page：792, May 2013, [Reviewed], [Lead, Corresponding], ［International magazine］
  Coordinated proliferation between clonally distinct cells via inter-cell-layer signaling largely determines the size and shape of plant organs. Nonetheless, the signaling mechanism underlying this coordination in leaves remains elusive because of a lack of understanding of the signaling molecule (or molecules) involved. ANGUSTIFOLIA3 (AN3, also called GRF-INTERACTING FACTOR1) encodes a putative transcriptional coactivator with homology to human synovial sarcoma translocation protein. AN3 transcripts accumulate in mesophyll cells but are not detectable in leaf epidermal cells. However, we found here that in addition to mesophyll cells, epidermal cells of an3 leaves show defective proliferation. This spatial difference between the accumulation pattern of AN3 transcripts and an3 leaf phenotype is explained by AN3 protein movement across cell layers. AN3 moves into epidermal cells after being synthesized within mesophyll cells and helps control epidermal cell proliferation. Interference with AN3 movement results in abnormal leaf size and shape, indicating that AN3 signaling is indispensable for normal leaf development. AN3 movement does not require type II chaperonin activity, which is needed for movement of some mobile proteins. Taking these findings together, we present a novel model emphasizing the role of mesophyll cells as a signaling source coordinating proliferation between clonally independent leaf cells.
  Elsevier {BV}, English, Scientific journal
  DOI：https://doi.org/10.1016/j.cub.2013.03.044
  DOI ID：10.1016/j.cub.2013.03.044, ORCID：33951052, PubMed ID：23602479
• Key Proliferative Activity in the Junction between the Leaf Blade and Leaf Petiole of Arabidopsis　　　　　　　　　　　　　　　
  Yasunori Ichihashi; Kensuke Kawade; Takeshi Usami; Gorou Horiguchi; Taku Takahashi; Hirokazu Tsukaya
  PLANT PHYSIOLOGY, Volume：157, Number：3, First page：1151, Last page：1162, Aug. 2011, [Reviewed]
  American Society of Plant Biologists ({ASPB}), Scientific journal
  DOI：https://doi.org/10.1104/pp.111.185066
  DOI ID：10.1104/pp.111.185066, ORCID：33951187
• Non-cell-autonomously coordinated organ size regulation in leaf development　　　　　　　　　　　　　　　
  Kensuke Kawade; Gorou Horiguchi; Hirokazu Tsukaya
  Development, Volume：137, Number：24, First page：4221, Last page：4227, Nov. 2010, [Reviewed], [Lead]
  The Company of Biologists, Scientific journal
  DOI：https://doi.org/10.1242/dev.057117
  DOI ID：10.1242/dev.057117, CiNii Articles ID：20001449881, ORCID：33951121
• Transcriptional control of two ribosome-inactivating protein genes expressed in spinach (Spinacia oleracea) embryos　　　　　　　　　　　　　　　
  Kensuke Kawade; Kiyoshi Masuda
  Plant Physiology and Biochemistry, Volume：47, Number：5, First page：327, Last page：334, May 2009, [Reviewed], [Lead, Corresponding]
  Elsevier {BV}, Scientific journal
  DOI：https://doi.org/10.1016/j.plaphy.2008.12.020
  DOI ID：10.1016/j.plaphy.2008.12.020, ORCID：33950839
• Differential expression of ribosome-inactivating protein genes during somatic embryogenesis in spinach (Spinacia oleracea)　　　　　　　　　　　　　　　
  Kensuke Kawade; Takuma Ishizaki; Kiyoshi Masuda
  Physiologia Plantarum, Volume：134, Number：2, First page：270, Last page：281, Oct. 2008, [Reviewed], [Lead]
  Wiley-Blackwell, Scientific journal
  DOI：https://doi.org/10.1111/j.1399-3054.2008.01129.x
  DOI ID：10.1111/j.1399-3054.2008.01129.x, ORCID：33950837

• コケ植物における茎葉形成に重要なアルギニン代謝　　　　　　　　　　　　　　　
  川出健介
  Volume：79, First page：222, Last page：223, 2021, [Lead, Corresponding]
• 代謝システムからみる植物の形づくり　　　　　　　　　　　　　　　
  川出健介
  Volume：3, First page：453, Last page：457, 2019, [Lead, Corresponding]
• 表皮細胞の大きさと核内倍加のサイコロゲーム　　　　　　　　　　　　　　　
  川出健介
  2019, [Lead, Corresponding]
• 拡散により植物モルフォゲンの濃度勾配ができる仕組み　　　　　　　　　　　　　　　
  川出健介; 谷本博一
  2017, [Lead, Corresponding]

• マメ科植物における根粒共生と水分獲得のトレードオフ　　　　　　　　　　　　　　　
  川出健介
  Oct. 2024, [Invited]
  Oct. 2024 - Oct. 2024
• ヒメツリガネゴケの体制とアルギニン代謝の流れる方向性　　　　　　　　　　　　　　　
  川出健介
  2023, [Invited]
• 植物の幹細胞活性とアルギニン代謝を紐付ける仕組み　　　　　　　　　　　　　　　
  川出健介
  2019, [Invited]
• 発生シグナルの時空間動態を解くための実験と理論　　　　　　　　　　　　　　　
  川出健介
  2018, [Invited]
• Biased distribution of tissue-scale diffusivity shapes ANGUSTIFOLIA3 signaling gradient in growing leaf tissue　　　　　　　　　　　　　　　
  Kensuke Kawade
  2017, [Invited]
• 組織内の偏った拡散性が細胞間シグナル因子の発現勾配を形づくる　　　　　　　　　　　　　　　
  川出健介
  2017, [Invited]
• Plant morphogen intertwined with metabolism　　　　　　　　　　　　　　　
  Kensuke Kawade
  2017, [Invited]
• Spatiotemporal control of cell proliferation activity in a heterogeneous field of protein diffusivity in plants　　　　　　　　　　　　　　　
  Kensuke Kawade
  2014, [Invited]
• Cell-to-cell variability of protein trafficking dynamics in leaf primordia　　　　　　　　　　　　　　　
  Kensuke Kawade
  2014, [Invited]
• 葉原基におけるタンパク質拡散係数の非一様性と細胞増殖活性の時空間分布　　　　　　　　　　　　　　　
  川出健介
  2013, [Invited]
• 葉原基におけるAN3 の発現勾配と細胞増殖活性の時空間分布　　　　　　　　　　　　　　　
  川出健介
  2013, [Invited]

• Transcale Systemic Signaling and Metabolomics for Symbiosis　　　　　　　　　　　　　　　
  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (A), Apr. 2023 - Mar. 2027
  National Institute for Basic Biology
  Grant amount(Total)：47060000, Direct funding：36200000, Indirect funding：10860000
  Grant number：23H00381
• Novel role for arginine metabolism in the evolution of plant body plans　　　　　　　　　　　　　　　
  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Apr. 2022 - Mar. 2025
  National Institute for Basic Biology
  Grant amount(Total)：4290000, Direct funding：3300000, Indirect funding：990000
  Grant number：22K06289
• Deciphering developmental signal intertwined with primary metabolism by trans-omics approach　　　　　　　　　　　　　　　
  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B), Grant-in-Aid for Young Scientists (B), 01 Apr. 2017 - 31 Mar. 2020
  Kawade Kensuke
  Grant amount(Total)：4550000, Direct funding：3500000, Indirect funding：1050000
  Pattern formation of organisms requires cell proliferation and expansion, indicating that there are mechanisms, which connect tissue patterning with metabolic reprogramming to stimulate these cellular processes. However, it is unclear how metabolism is modified in response to cell proliferation and expansion. We here addressed this question using plant seedlings as a model. One important finding is an identification of a mechanism for cell proliferation mediated by a regulation of amino acid metabolism. Moreover, we clarified that the integration between tissue patterning and amino acid metabolism has been already established in an early land plant. Interestingly, in the early land plant, this mechanism may facilitate complex tissue patterning including shoot formation because we observed stunted and simple shoot morphology when disrupted. These results thus augmented our understanding in mechanisms by which developmental and metabolic processes are integrated for tissue patterning.
  Grant number：17K15147
• Elucidation of metabolic network underlying plant development　　　　　　　　　　　　　　　
  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Jun. 2013 - Mar. 2018
  Hirai Masami; SAWADA Yuji; LI Yimeng; SAKAI Yuji; SRIYUDTHSAK Kansuporn; OKAMURA Eiji; KAWADE Kensuke; VERGARA Fredd, Institute of Physical and Chemical Research
  Grant amount(Total)：141570000, Direct funding：108900000, Indirect funding：32670000
  Metabolism is a basis of life, but its dynamics and regulatory function in development have not been fully understood. In this study, we aimed to understand the dynamics of plant metabolic system via a mathematical approach, and to identify metabolic pathways that regulate developmental processes by molecular genetics. We performed integrated mathematical and biological studies, i.e., theory construction and its application, and, prediction and confirmation, which have led to understanding of several developmental processes. We also clarified by a molecular genetic approach that a metabolic enzyme CYP77A4-mediated metabolic pathway regulates patterning of Arabidopsis embryo.
  Grant number：25113010
• Multidimensional Exploration of Logics of Leaf Development　　　　　　　　　　　　　　　
  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Jun. 2013 - Mar. 2018
  Tsukaya Hirokazu; SAKAKIBARA Keiko; YAMAGUCHI Takahiro; KAWADE Kensuke; Koga Hiroyuki, The University of Tokyo
  Grant amount(Total)：165750000, Direct funding：127500000, Indirect funding：38250000
  We could substitute past knowledge or add a new topic to the present textbooks during this research project. Namely, (1) the relationship between the ploidy level regulated by endoredupication and cell size is not as simple as formerly believed (Tsukaya 2013; Katagiri et al. 2016: the latter received The Hirase Award from The Japanese Society of Plant Morphology); (2) spatial-temporal regulation of leaf meristem can be explained by a classic morphogen-like diffusion of AN3 protein (Kawade et al. 2017).
  Grant number：25113002
• Biased distribution of protein diffusivity in leaf primordia　　　　　　　　　　　　　　　
  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B), Grant-in-Aid for Young Scientists (B), 01 Apr. 2015 - 31 Mar. 2017
  KAWADE Kensuke, Okazaki Research Facilities, National Institutes of Natural Sciences
  Grant amount(Total)：3120000, Direct funding：2400000, Indirect funding：720000
  We experimentally and theoretically studied how AN3 signaling gradient is formed along leaf proximal-to-distal axis, and how this gradient regulates the spatiotemporal dynamics of cell proliferation in leaves. We succeeded in establishing a quantitative model, which well summarize the AN3 gradient formation in a realistic spatial field. This model predicted that differential distribution of cell size has impact to determine the signaling gradient. Based on this finding, we proposed that biased distribution of tissue-scale diffusivity is a core mechanism to achieve the AN3 signaling gradient; hence cell proliferation dynamics in leaves. We reported these results in The Japanese Society of Plant Morphology and The Botanical Society of Japan. Now, we are submitting this to an international journal for publication (Kawade et al., submitted).
  Grant number：15K18549
• 葉原基におけるＡＮ３の発現勾配と細胞増殖活性の時空間分布　　　　　　　　　　　　　　　
  25 Apr. 2014 - 31 Mar. 2015
  Grant amount(Total)：1820000, Direct funding：1400000, Indirect funding：420000
  Grant number：14J02337
• CRE/Loxシステムを用いたキメラ解析による葉器官サイズ制御機構の解明　　　　　　　　　　　　　　　
  2009 - 2010
  Grant amount(Total)：1400000, Direct funding：1400000
  Grant number：09J07214

• プレスリリース「植物の形づくりを促すアミノ酸代謝を発見」　　　　　　　　　　　　　　　
  Myself, Sep. 2020
• プレスリリース「植物の双葉を２枚にする酵素を発見」　　　　　　　　　　　　　　　
  Myself, Sep. 2018
• プレスリリース「シンプルな物理現象「拡散」で葉の形や大きさが決まる仕組みを解明」　　　　　　　　　　　　　　　
  Myself, Sep. 2017
• プレスリリース「植物細胞の大きさを決めるサイコロゲームを発見」　　　　　　　　　　　　　　　
  Myself, Sep. 2017
• プレスリリース「組織間での情報伝達を介した葉の成長メカニズムを解明 ‒農作物の収量 増産など応用分野に期待‒」　　　　　　　　　　　　　　　
  Myself, Apr. 2013
• プレスリリース「葉の大きさは細胞間のコミュニケーションにより制御される」　　　　　　　　　　　　　　　
  Myself, Nov. 2010