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Prof. Dr. Kazuyuki Kuchitsu
Department of Industrial Administration, Tokyo University of Science, 2641 Yamazaki, Noda-shi, Chiba-ken 278-8510, Japan

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Research Keywords & Expertise

0 Cell Biology
0 Plant Physiology
0 Signal Transduction
0 cell death
0 Plant molecular biology

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cell death
Signal Transduction
Molecular mechanisms for Ca2+-mediated signal transduction in rice
plant immunity

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Journal article
Published: 29 March 2021 in Plants
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Inhibitors of human two-pore channels (TPC1 and TPC2), i.e., verapamil, tetrandrine, and NED-19, are promising medicines used in treatment of serious diseases. In the present study, the impact of these substances on action potentials (APs) and vacuolar channel activity was examined in the aquatic characean algae Nitellopsis obtusa and in the terrestrial liverwort Marchantia polymorpha. In both plant species, verapamil (20–300 µM) caused reduction of AP amplitudes, indicating impaired Ca2+ transport. In N. obtusa, it depolarized the AP excitation threshold and resting potential and prolonged AP duration. In isolated vacuoles of M. polymorpha, verapamil caused a reduction of the open probability of slow vacuolar SV/TPC channels but had almost no effect on K+ channels in the tonoplast of N. obtusa. In both species, tetrandrine (20–100 µM) evoked a pleiotropic effect: reduction of resting potential and AP amplitudes and prolongation of AP repolarization phases, especially in M. polymorpha, but it did not alter vacuolar SV/TPC activity. NED-19 (75 µM) caused both specific and unspecific effects on N. obtusa APs. In M. polymorpha, NED-19 increased the duration of repolarization. However, no inhibition of SV/TPC channels was observed in Marchantia vacuoles, but an increase in open probability and channel flickering. The results indicate an effect on Ca2+ -permeable channels governing plant excitation.

ACS Style

Mateusz Koselski; Vilmantas Pupkis; Kenji Hashimoto; Indre Lapeikaite; Agnieszka Hanaka; Piotr Wasko; Egle Plukaite; Kazuyuki Kuchitsu; Vilma Kisnieriene; Kazimierz Trebacz. Impact of Mammalian Two-Pore Channel Inhibitors on Long-Distance Electrical Signals in the Characean Macroalga Nitellopsis obtusa and the Early Terrestrial Liverwort Marchantia polymorpha. Plants 2021, 10, 647 .

AMA Style

Mateusz Koselski, Vilmantas Pupkis, Kenji Hashimoto, Indre Lapeikaite, Agnieszka Hanaka, Piotr Wasko, Egle Plukaite, Kazuyuki Kuchitsu, Vilma Kisnieriene, Kazimierz Trebacz. Impact of Mammalian Two-Pore Channel Inhibitors on Long-Distance Electrical Signals in the Characean Macroalga Nitellopsis obtusa and the Early Terrestrial Liverwort Marchantia polymorpha. Plants. 2021; 10 (4):647.

Chicago/Turabian Style

Mateusz Koselski; Vilmantas Pupkis; Kenji Hashimoto; Indre Lapeikaite; Agnieszka Hanaka; Piotr Wasko; Egle Plukaite; Kazuyuki Kuchitsu; Vilma Kisnieriene; Kazimierz Trebacz. 2021. "Impact of Mammalian Two-Pore Channel Inhibitors on Long-Distance Electrical Signals in the Characean Macroalga Nitellopsis obtusa and the Early Terrestrial Liverwort Marchantia polymorpha." Plants 10, no. 4: 647.

Paper
Published: 14 December 2020 in The Analyst
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Stimulated Raman scattering microscopy allows for multicolour label-free chemical imaging of plant tissues.

ACS Style

Takanori Iino; Kenji Hashimoto; Takuya Asai; Kazuyuki Kuchitsu; Yasuyuki Ozeki. Multicolour chemical imaging of plant tissues with hyperspectral stimulated Raman scattering microscopy. The Analyst 2020, 146, 1234 -1238.

AMA Style

Takanori Iino, Kenji Hashimoto, Takuya Asai, Kazuyuki Kuchitsu, Yasuyuki Ozeki. Multicolour chemical imaging of plant tissues with hyperspectral stimulated Raman scattering microscopy. The Analyst. 2020; 146 (4):1234-1238.

Chicago/Turabian Style

Takanori Iino; Kenji Hashimoto; Takuya Asai; Kazuyuki Kuchitsu; Yasuyuki Ozeki. 2020. "Multicolour chemical imaging of plant tissues with hyperspectral stimulated Raman scattering microscopy." The Analyst 146, no. 4: 1234-1238.

Journal article
Published: 01 December 2020 in International Journal of Molecular Sciences
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Autophagy is ubiquitous in eukaryotic cells and plays an essential role in stress adaptation and development by recycling nutrients and maintaining cellular homeostasis. However, the dynamics and regulatory mechanisms of autophagosome formation during the cell cycle in plant cells remain poorly elucidated. We here analyzed the number of autophagosomes during cell cycle progression in synchronized tobacco BY-2 cells expressing YFP-NtATG8a as a marker for the autophagosomes. Autophagosomes were abundant in the G2 and G1 phases of interphase, though they were much less abundant in the M and S phases. Autophagosomes drastically decreased during the G2/M transition, and the CDK inhibitor roscovitine inhibited the G2/M transition and the decrease in autophagosomes. Autophagosomes were rapidly increased by a proteasome inhibitor, MG-132. MG-132-induced autophagosome formation was also markedly lower in the M phases than during interphase. These results indicate that the activity of autophagosome formation is differently regulated at each cell cycle stage, which is strongly suppressed during mitosis.

ACS Style

Shigeru Hanamata; Takamitsu Kurusu; Kazuyuki Kuchitsu. Cell Cycle-Dependence of Autophagic Activity and Inhibition of Autophagosome Formation at M Phase in Tobacco BY-2 Cells. International Journal of Molecular Sciences 2020, 21, 9166 .

AMA Style

Shigeru Hanamata, Takamitsu Kurusu, Kazuyuki Kuchitsu. Cell Cycle-Dependence of Autophagic Activity and Inhibition of Autophagosome Formation at M Phase in Tobacco BY-2 Cells. International Journal of Molecular Sciences. 2020; 21 (23):9166.

Chicago/Turabian Style

Shigeru Hanamata; Takamitsu Kurusu; Kazuyuki Kuchitsu. 2020. "Cell Cycle-Dependence of Autophagic Activity and Inhibition of Autophagosome Formation at M Phase in Tobacco BY-2 Cells." International Journal of Molecular Sciences 21, no. 23: 9166.

Original research article
Published: 06 March 2020 in Frontiers in Plant Science
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Autophagy has recently been shown to be required for tapetal programmed cell death (PCD) and pollen maturation in rice. A transcriptional regulatory network is also known to play a key role in the progression of tapetal PCD. However, the relationship between the gene regulatory network and autophagy in rice anther development is mostly unknown. Here, we comprehensively analyzed the effect of autophagy disruption on gene expression profile during the tapetal PCD in rice anther development using high-throughput RNA sequencing. Expression of thousands of genes, including specific transcription factors and several proteases required for tapetal degradation, fluctuated synchronously at specific stages during tapetal PCD progression in the wild-type anthers, while this fluctuation showed significant delay in the autophagy-deficient mutant Osatg7-1. Moreover, gene ontology enrichment analysis in combination with self-organizing map clustering as well as pathway analysis revealed that the expression patterns of a variety of organelle-related genes as well as genes involved in carbohydrate/lipid metabolism were affected in the Osatg7-1 mutant during pollen maturation. These results suggest that autophagy is required for proper regulation of gene expression and quality control of organelles and timely progression of tapetal PCD during rice pollen development.

ACS Style

Shigeru Hanamata; Jumpei Sawada; Seijiro Ono; Kazunori Ogawa; Togo Fukunaga; Ken–Ichi Nonomura; Seisuke Kimura; Takamitsu Kurusu; Kazuyuki Kuchitsu. Impact of Autophagy on Gene Expression and Tapetal Programmed Cell Death During Pollen Development in Rice. Frontiers in Plant Science 2020, 11, 172 .

AMA Style

Shigeru Hanamata, Jumpei Sawada, Seijiro Ono, Kazunori Ogawa, Togo Fukunaga, Ken–Ichi Nonomura, Seisuke Kimura, Takamitsu Kurusu, Kazuyuki Kuchitsu. Impact of Autophagy on Gene Expression and Tapetal Programmed Cell Death During Pollen Development in Rice. Frontiers in Plant Science. 2020; 11 ():172.

Chicago/Turabian Style

Shigeru Hanamata; Jumpei Sawada; Seijiro Ono; Kazunori Ogawa; Togo Fukunaga; Ken–Ichi Nonomura; Seisuke Kimura; Takamitsu Kurusu; Kazuyuki Kuchitsu. 2020. "Impact of Autophagy on Gene Expression and Tapetal Programmed Cell Death During Pollen Development in Rice." Frontiers in Plant Science 11, no. : 172.

Journal article
Published: 06 December 2019 in Scientific Reports
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Autophagy plays crucial roles in the recycling of metabolites, and is involved in many developmental processes. Rice mutants defective in autophagy are male sterile due to immature pollens, indicating its critical role in pollen development. However, physiological roles of autophagy during seed maturation had remained unknown. We here found that seeds of the rice autophagy-deficient mutant Osatg7-1, that produces seeds at a very low frequency in paddy fields, are smaller and show chalky appearance and lower starch content in the endosperm at the mature stage under normal growth condition. We comprehensively analyzed the effects of disruption of autophagy on biochemical properties, proteome and seed quality, and found an abnormal activation of starch degradation pathways including accumulation of α-amylases in the endosperm during seed maturation in Osatg7-1. These results indicate critical involvement of autophagy in metabolic regulation in the endosperm of rice, and provide insights into novel autophagy-mediated regulation of starch metabolism during seed maturation.

ACS Style

Yuri Sera; Shigeru Hanamata; Shingo Sakamoto; Seijiro Ono; Kentaro Kaneko; Yuudai Mitsui; Tomoko Koyano; Naoko Fujita; Ai Sasou; Takehiro Masumura; Hikaru Saji; Ken-Ichi Nonomura; Nobutaka Mitsuda; Toshiaki Mitsui; Takamitsu Kurusu; Kazuyuki Kuchitsu. Essential roles of autophagy in metabolic regulation in endosperm development during rice seed maturation. Scientific Reports 2019, 9, 1 -14.

AMA Style

Yuri Sera, Shigeru Hanamata, Shingo Sakamoto, Seijiro Ono, Kentaro Kaneko, Yuudai Mitsui, Tomoko Koyano, Naoko Fujita, Ai Sasou, Takehiro Masumura, Hikaru Saji, Ken-Ichi Nonomura, Nobutaka Mitsuda, Toshiaki Mitsui, Takamitsu Kurusu, Kazuyuki Kuchitsu. Essential roles of autophagy in metabolic regulation in endosperm development during rice seed maturation. Scientific Reports. 2019; 9 (1):1-14.

Chicago/Turabian Style

Yuri Sera; Shigeru Hanamata; Shingo Sakamoto; Seijiro Ono; Kentaro Kaneko; Yuudai Mitsui; Tomoko Koyano; Naoko Fujita; Ai Sasou; Takehiro Masumura; Hikaru Saji; Ken-Ichi Nonomura; Nobutaka Mitsuda; Toshiaki Mitsui; Takamitsu Kurusu; Kazuyuki Kuchitsu. 2019. "Essential roles of autophagy in metabolic regulation in endosperm development during rice seed maturation." Scientific Reports 9, no. 1: 1-14.

Journal article
Published: 25 June 2019 in Plant Biotechnology
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We have previously shown that autophagy is required for post meiotic anther development including programmed cell death-mediated degradation of the tapetum and pollen maturation in rice. However, the spatiotemporal dynamics of autophagy in the tapetum remain poorly understood. We here established an in vivo imaging technique to analyze the dynamics of autophagy in rice tapetum cells by expressing green fluorescent protein-tagged AtATG8, a marker for autophagosomes. 3D-imaging analysis revealed that the number of autophagosomes/autophagy-related structures is extremely low at the tetrad stage (stage 8), and autophagy is dramatically induced at the uninucleate stages (stage 9-10) throughout the tapetal cells during anther development. The present monitoring system for autophagy offers a powerful tool to analyze the regulation of autophagy in rice tapetal cells during pollen maturation.

ACS Style

Shigeru Hanamata; Jumpei Sawada; Bunki Toh; Seijiro Ono; Kazunori Ogawa; Togo Fukunaga; Ken-Ichi Nonomura; Takamitsu Kurusu; Kazuyuki Kuchitsu. Monitoring autophagy in rice tapetal cells during pollen maturation. Plant Biotechnology 2019, 36, 99 -105.

AMA Style

Shigeru Hanamata, Jumpei Sawada, Bunki Toh, Seijiro Ono, Kazunori Ogawa, Togo Fukunaga, Ken-Ichi Nonomura, Takamitsu Kurusu, Kazuyuki Kuchitsu. Monitoring autophagy in rice tapetal cells during pollen maturation. Plant Biotechnology. 2019; 36 (2):99-105.

Chicago/Turabian Style

Shigeru Hanamata; Jumpei Sawada; Bunki Toh; Seijiro Ono; Kazunori Ogawa; Togo Fukunaga; Ken-Ichi Nonomura; Takamitsu Kurusu; Kazuyuki Kuchitsu. 2019. "Monitoring autophagy in rice tapetal cells during pollen maturation." Plant Biotechnology 36, no. 2: 99-105.

Review
Published: 04 March 2019 in Plant Signaling & Behavior
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Inter-cellular and inter-kingdom signaling systems of various levels of complexity regulate pathogenic and mutualistic interactions between bacteria, parasites, and fungi and animal and plant hosts. Inter-kingdom interactions between mutualistic bacteria such as rhizobia and legumes during nodulation and between fungi and plants during mycorrhizal associations, are characterized by the extensive exchange of molecular signals, which allow nitrogen and phosphate assimilation, respectively. A novel aspect of this signaling exchange is the existence of specific structures, the exosomes, that carry important molecules that shape the plant–pathogen interactions. Exosomes contain a wide array of molecules, such as lipids, proteins, messenger RNA, and microRNAs, that play important roles in cell-to-cell communication in animal and plant cells by affecting gene expression and other physiological activity in distant cells within the same organism (e.g., during cancer metastases and neuron injuries). In plant cells, it has been recently reported that exosomes go beyond organism boundaries and inhibit a pathogenic interaction in plants. Plant produce and send exosomes loaded with specific small miRNA which inhibit the pathogen infection, but the pathogen can also produce exosomes carrying pro-pathogenic proteins and microRNAs. Therefore, exosomes are the important bridge regulating the signal exchange. Exosomes are small membrane-bound vesicles derived from multivesicular bodies (MVBs), which carries selected cargos from the cytoplasm (protein, lipids, and microRNAs) and under certain circumstances, they fuse with the plasma membrane, releasing the small vesicles as cargo-carrying exosomes into the extracellular space during intercellular and inter-kingdom communication. Animal and plant proteomic studies have demonstrated that tetraspanin proteins are an integral part of exosome membranes, positioning tetraspanins as essential components for endosome organization, with key roles in membrane fusion, cell trafficking, and membrane recognition. We discuss the similarities and differences between animal tetraspanins and plant tetraspanins formed during plant–microbe interactions and their potential role in mutualistic communication.

ACS Style

Saul Jimenez-Jimenez; Kenji Hashimoto; Olivia Santana; Jesús Aguirre; Kazuyuki Kuchitsu; Luis Cárdenas. Emerging roles of tetraspanins in plant inter-cellular and inter-kingdom communication. Plant Signaling & Behavior 2019, 14, e1581559 .

AMA Style

Saul Jimenez-Jimenez, Kenji Hashimoto, Olivia Santana, Jesús Aguirre, Kazuyuki Kuchitsu, Luis Cárdenas. Emerging roles of tetraspanins in plant inter-cellular and inter-kingdom communication. Plant Signaling & Behavior. 2019; 14 (4):e1581559.

Chicago/Turabian Style

Saul Jimenez-Jimenez; Kenji Hashimoto; Olivia Santana; Jesús Aguirre; Kazuyuki Kuchitsu; Luis Cárdenas. 2019. "Emerging roles of tetraspanins in plant inter-cellular and inter-kingdom communication." Plant Signaling & Behavior 14, no. 4: e1581559.

Original article
Published: 20 December 2018 in The Plant Journal
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Reactive oxygen species (ROS) produced by NADPH oxidases, respiratory burst oxidase homologs (Rbohs), play crucial roles in development as well as biotic and abiotic stress responses in plants. Arabidopsis has ten Rboh genes, AtRbohA to AtRbohJ. Five AtRbohs (AtRbohC, D, F, H and J) are synergistically activated by Ca2+ and protein phosphorylation to produce ROS that play various roles in planta, although the activities of the other Rbohs remain unknown. With a heterologous expression system, we found a range in ROS–producing activity among the AtRbohs with differences up to 100 times, indicating that the required ROS amount is different in each situation where the AtRbohs act. To specify the functions of AtRbohs involved in cell growth, we focused on AtRbohC, H and J, which are involved in tip growth of root hairs or pollen tubes. Ectopic expression of the root hair factor AtRbohC/ROOT HAIR DEFECTIVE 2 (RHD2) in pollen tubes restored the atrbohH atrbohJ defects in tip growth of pollen tubes. However, expression of AtRbohH or J in root hairs did not complement the tip growth defect in the atrbohC/rhd2 mutant. Our data indicate that Rbohs possess different ranges of enzymatic activity, and that some Rbohs have evolved to carry specific functions in cell growth. This article is protected by copyright. All rights reserved.

ACS Style

Hidetaka Kaya; Seiji Takeda; Masaki Kobayashi; Sachie Kimura; Ayako Iizuka; Aya Imai; Haruka Hishinuma; Tomoko Kawarazaki; Kyoichiro Mori; Yuta Yamamoto; Yuki Murakami; Ayuko Nakauchi; Mitsutomo Abe; Kazuyuki Kuchitsu. Comparative analysis of the reactive oxygen species‐producing enzymatic activity of Arabidopsis NADPH oxidases. The Plant Journal 2018, 98, 291 -300.

AMA Style

Hidetaka Kaya, Seiji Takeda, Masaki Kobayashi, Sachie Kimura, Ayako Iizuka, Aya Imai, Haruka Hishinuma, Tomoko Kawarazaki, Kyoichiro Mori, Yuta Yamamoto, Yuki Murakami, Ayuko Nakauchi, Mitsutomo Abe, Kazuyuki Kuchitsu. Comparative analysis of the reactive oxygen species‐producing enzymatic activity of Arabidopsis NADPH oxidases. The Plant Journal. 2018; 98 (2):291-300.

Chicago/Turabian Style

Hidetaka Kaya; Seiji Takeda; Masaki Kobayashi; Sachie Kimura; Ayako Iizuka; Aya Imai; Haruka Hishinuma; Tomoko Kawarazaki; Kyoichiro Mori; Yuta Yamamoto; Yuki Murakami; Ayuko Nakauchi; Mitsutomo Abe; Kazuyuki Kuchitsu. 2018. "Comparative analysis of the reactive oxygen species‐producing enzymatic activity of Arabidopsis NADPH oxidases." The Plant Journal 98, no. 2: 291-300.

Chapter
Published: 05 October 2018 in Studies on Biomarkers and New Targets in Aging Research in Iran
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Control of freezing in plant tissues is a key issue in cold hardiness mechanisms. Yet freeze-regulation mechanisms remain mostly unexplored. Among them, ice nucleation activity (INA) is a primary factor involved in the initiation and regulation of freezing events in plant tissues, yet the details remain poorly understood. To address this, we developed a highly reproducible assay for determining plant tissue INA and noninvasive freeze visualization tools using MRI and infrared thermography. The results of visualization studies on plant freezing behaviors and INA survey of over 600 species tissues show that (1) freezing-sensitive plants tend to have low INA in their tissues (thus tend to transiently supercool), while wintering cold-hardy species have high INA in some specialized tissues; and (2) the high INA in cold-hardy tissues likely functions as a freezing sensor to initiate freezing at warm subzero temperatures at appropriate locations and timing, resulting in the induction of tissue-/species-specific freezing behaviors (e.g., extracellular freezing, extraorgan freezing) and the freezing order among tissues: from the primary freeze to the last tissue remaining unfrozen (likely INA level dependent). The spatiotemporal distributions of tissue INA, their characterization, and functional roles are detailed. INA assay principles, anti-nucleation activity (ANA), and freeze visualization tools are also described.

ACS Style

Masaya Ishikawa; Hideyuki Yamazaki; Tadashi Kishimoto; Hiroki Murakawa; Timothy Stait-Gardner; Kazuyuki Kuchitsu; William S. Price. Ice Nucleation Activity in Plants: The Distribution, Characterization, and Their Roles in Cold Hardiness Mechanisms. Studies on Biomarkers and New Targets in Aging Research in Iran 2018, 1081, 99 -115.

AMA Style

Masaya Ishikawa, Hideyuki Yamazaki, Tadashi Kishimoto, Hiroki Murakawa, Timothy Stait-Gardner, Kazuyuki Kuchitsu, William S. Price. Ice Nucleation Activity in Plants: The Distribution, Characterization, and Their Roles in Cold Hardiness Mechanisms. Studies on Biomarkers and New Targets in Aging Research in Iran. 2018; 1081 ():99-115.

Chicago/Turabian Style

Masaya Ishikawa; Hideyuki Yamazaki; Tadashi Kishimoto; Hiroki Murakawa; Timothy Stait-Gardner; Kazuyuki Kuchitsu; William S. Price. 2018. "Ice Nucleation Activity in Plants: The Distribution, Characterization, and Their Roles in Cold Hardiness Mechanisms." Studies on Biomarkers and New Targets in Aging Research in Iran 1081, no. : 99-115.

Research letter
Published: 26 June 2018 in FEBS Letters
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Root hairs (RH) are tip growing polarized cells aiding the uptake of nutrients and water into plants. RH differentiation involves the interplay of various hormones and second messengers. Tightly regulated production of reactive oxygen species by the NADPH oxidase RBOHC crucially functions in RH differentiation and Ca2+‐dependent phosphorylation has been implemented in these processes. However, the kinases regulating RBOHC remained enigmatic. Here we identify CBL1‐CIPK26 Ca2+ sensor‐kinase complexes as modulators of RBOHC activity. Combined genetic, cell biological and biochemical analyses reveal synergistic function of CIPK26‐mediated phosphorylation and Ca2+ binding for RBOHC activation. Complementation of rbohC mutant RH phenotypes by a S318/322 phosphorylation deficient RBOHC version suggests flexible and alternating phosphorylation patterns as mechanism fine‐tuning ROS production in RH development.

ACS Style

Xinxin Zhang; Philipp Köster; Kathrin Schlücking; Daria Balcerowicz; Kenji Hashimoto; Kazuyuki Kuchitsu; Kris Vissenberg; Jörg Kudla. CBL1‐CIPK26‐mediated phosphorylation enhances activity of the NADPH oxidase RBOHC, but is dispensable for root hair growth. FEBS Letters 2018, 592, 2582 -2593.

AMA Style

Xinxin Zhang, Philipp Köster, Kathrin Schlücking, Daria Balcerowicz, Kenji Hashimoto, Kazuyuki Kuchitsu, Kris Vissenberg, Jörg Kudla. CBL1‐CIPK26‐mediated phosphorylation enhances activity of the NADPH oxidase RBOHC, but is dispensable for root hair growth. FEBS Letters. 2018; 592 (15):2582-2593.

Chicago/Turabian Style

Xinxin Zhang; Philipp Köster; Kathrin Schlücking; Daria Balcerowicz; Kenji Hashimoto; Kazuyuki Kuchitsu; Kris Vissenberg; Jörg Kudla. 2018. "CBL1‐CIPK26‐mediated phosphorylation enhances activity of the NADPH oxidase RBOHC, but is dispensable for root hair growth." FEBS Letters 592, no. 15: 2582-2593.

Short communication
Published: 04 May 2018 in Communicative & Integrative Biology
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Pharmacological indications suggest that anion channel-mediated plasma membrane (PM) anion efflux is crucial in early defense signaling to induce immune responses and programmed cell death in plants. Arabidopsis SLAC1, an S-type anion channel required for stomatal closure, is involved in cryptogein-induced PM Cl− efflux to positively modulate the activation of other ion fluxes, production of reactive oxygen species and a wide range of defense responses including hypersensitive cell death in tobacco BY-2 cells. We here analyzed disease resistance against several pathogens in multiple mutants of the SLAC/SLAH channels of Arabidopsis. Resistance against a biotrophic oomycete Hyaloperonospora arabidopsidis Noco2 was significantly enhanced in the SLAC1-overexpressing plants than in the wild-type, while that against a bacteria Pseudomonas syringae was not affected significantly. Possible regulatory roles of S-type anion channels in plant immunity and disease resistance against bacterial and oomycete pathogens is discussed.

ACS Style

Takamitsu Kurusu; Daiki Mitsuka; Chikako Yagi; Nobutaka Kitahata; Tomokazu Tsutsui; Takashi Ueda; Yoshiko Yamamoto; Juntaro Negi; Koh Iba; Shigeyuki Betsuyaku; Kazuyuki Kuchitsu. Involvement of S-type anion channels in disease resistance against an oomycete pathogen in Arabidopsis seedlings. Communicative & Integrative Biology 2018, 11, 1 -6.

AMA Style

Takamitsu Kurusu, Daiki Mitsuka, Chikako Yagi, Nobutaka Kitahata, Tomokazu Tsutsui, Takashi Ueda, Yoshiko Yamamoto, Juntaro Negi, Koh Iba, Shigeyuki Betsuyaku, Kazuyuki Kuchitsu. Involvement of S-type anion channels in disease resistance against an oomycete pathogen in Arabidopsis seedlings. Communicative & Integrative Biology. 2018; 11 (3):1-6.

Chicago/Turabian Style

Takamitsu Kurusu; Daiki Mitsuka; Chikako Yagi; Nobutaka Kitahata; Tomokazu Tsutsui; Takashi Ueda; Yoshiko Yamamoto; Juntaro Negi; Koh Iba; Shigeyuki Betsuyaku; Kazuyuki Kuchitsu. 2018. "Involvement of S-type anion channels in disease resistance against an oomycete pathogen in Arabidopsis seedlings." Communicative & Integrative Biology 11, no. 3: 1-6.

Journal article
Published: 16 October 2017 in Sustainability
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The agriculture sector is known to be the one of the major contributors to global greenhouse gas (GHG) emissions. At the same time, global climate changes have affected the agriculture sector. In order to strengthen the sustainable development of agriculture, it is important to promote environmentally friendly farming and simultaneously increase the economic value. To improve the productivity of agriculture, technical advancements have occurred. Among those, we have focused on CO2 treatment in cultivation. We aimed to clarify the effectiveness of the elevated CO2 treatment of spinach based on GHG emission and the economic value using the eco-efficiency score. We assumed that nutrition could represent the value of the vegetable. We measured weights, vitamin C, and CO2 emissions of elevated CO2 treatment and conventional production. We used life cycle assessment (LCA) to estimate CO2 emissions. CO2 emissions of a 100-g bouquet of spinach were estimated from agricultural inputs, farming, transport, and distribution center processes at a model spinach farm in Japan. CO2 emission of elevated CO2 treatment was 29.0 g-CO2, and was 49.0 g-CO2 for conventional production. The net weight of a bouquet of elevated CO2-treated spinach was 1.69-fold greater than that of conventional production. Vitamin C per 100 g spinach produced via elevated CO2 treatment was 15.1 mg, and that of conventional production was 13.5 mg on average. Finally, based on the above results, we assessed the eco-efficiency scores of the elevated CO2 treatment and conventional production of spinach, enabling integration of the nutritional value and the environmental impact. The score showed that elevated CO2 treatment (0.76) was 2.9-fold more efficient than conventional production (0.26). This study suggested that elevated CO2 treatment could enhance growth and nutritional value of spinach, and further contribute to CO2 reduction.

ACS Style

Yuna Seo; Keisuke Ide; Nobutaka Kitahata; Kazuyuki Kuchitsu; Kiyoshi Dowaki. Environmental Impact and Nutritional Improvement of Elevated CO2 Treatment: A Case Study of Spinach Production. Sustainability 2017, 9, 1854 .

AMA Style

Yuna Seo, Keisuke Ide, Nobutaka Kitahata, Kazuyuki Kuchitsu, Kiyoshi Dowaki. Environmental Impact and Nutritional Improvement of Elevated CO2 Treatment: A Case Study of Spinach Production. Sustainability. 2017; 9 (10):1854.

Chicago/Turabian Style

Yuna Seo; Keisuke Ide; Nobutaka Kitahata; Kazuyuki Kuchitsu; Kiyoshi Dowaki. 2017. "Environmental Impact and Nutritional Improvement of Elevated CO2 Treatment: A Case Study of Spinach Production." Sustainability 9, no. 10: 1854.

Journal article
Published: 01 October 2017 in Journal of Virology
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Primary infection of a plant with a pathogen that causes high accumulation of salicylic acid in the plant typically via a hypersensitive response confers enhanced resistance against secondary infection with a broad spectrum of pathogens, including viruses. This phenomenon is called systemic acquired resistance (SAR), which is a plant priming for adaption to repeated biotic stress. However, the molecular mechanisms of SAR-mediated enhanced inhibition, especially of virus infection, remain unclear. Here, we show that SAR against cucumber mosaic virus (CMV) in tobacco plants ( Nicotiana tabacum ) involves a calmodulin-like protein, rgs-CaM. We previously reported the antiviral function of rgs-CaM, which binds to and directs degradation of viral RNA silencing suppressors (RSSs), including CMV 2b, via autophagy. We found that rgs-CaM-mediated immunity is ineffective against CMV infection in normally growing tobacco plants but is activated as a result of SAR induction via salicylic acid signaling. We then analyzed the effect of overexpression of rgs-CaM on salicylic acid signaling. Overexpressed and ectopically expressed rgs-CaM induced defense reactions, including cell death, generation of reactive oxygen species, and salicylic acid signaling. Further analysis using a combination of the salicylic acid analogue benzo-(1,2,3)-thiadiazole-7-carbothioic acid S -methyl ester (BTH) and the Ca 2+ ionophore A23187 revealed that rgs-CaM functions as an immune receptor that induces salicylic acid signaling by simultaneously perceiving both viral RSS and Ca 2+ influx as infection cues, implying its autoactivation. Thus, secondary infection of SAR-induced tobacco plants with CMV seems to be effectively inhibited through 2b recognition and degradation by rgs-CaM, leading to reinforcement of antiviral RNA silencing and other salicylic acid-mediated antiviral responses. IMPORTANCE Even without an acquired immune system like that in vertebrates, plants show enhanced whole-plant resistance against secondary infection with pathogens; this so-called systemic acquired resistance (SAR) has been known for more than half a century and continues to be extensively studied. SAR-induced plants strongly and rapidly express a number of antibiotics and pathogenesis-related proteins targeted against secondary infection, which can account for enhanced resistance against bacterial and fungal pathogens but are not thought to control viral infection. This study showed that enhanced resistance against cucumber mosaic virus is caused by a tobacco calmodulin-like protein, rgs-CaM, which detects and counteracts the major viral virulence factor (RNA silencing suppressor) after SAR induction. rgs-CaM-mediated SAR illustrates the growth versus defense trade-off in plants, as it targets the major virulence factor only under specific biotic stress conditions, thus avoiding the cost of constitutive activation while reducing the damage from virus infection.

ACS Style

Eun Jin Jeon; Kazuki Tadamura; Taiki Murakami; Jun-Ichi Inaba; Bo Min Kim; Masako Sato; Go Atsumi; Kazuyuki Kuchitsu; Chikara Masuta; Kenji S. Nakahara. rgs-CaM Detects and Counteracts Viral RNA Silencing Suppressors in Plant Immune Priming. Journal of Virology 2017, 91, 1 .

AMA Style

Eun Jin Jeon, Kazuki Tadamura, Taiki Murakami, Jun-Ichi Inaba, Bo Min Kim, Masako Sato, Go Atsumi, Kazuyuki Kuchitsu, Chikara Masuta, Kenji S. Nakahara. rgs-CaM Detects and Counteracts Viral RNA Silencing Suppressors in Plant Immune Priming. Journal of Virology. 2017; 91 (19):1.

Chicago/Turabian Style

Eun Jin Jeon; Kazuki Tadamura; Taiki Murakami; Jun-Ichi Inaba; Bo Min Kim; Masako Sato; Go Atsumi; Kazuyuki Kuchitsu; Chikara Masuta; Kenji S. Nakahara. 2017. "rgs-CaM Detects and Counteracts Viral RNA Silencing Suppressors in Plant Immune Priming." Journal of Virology 91, no. 19: 1.

Short communication
Published: 02 September 2017 in Plant Signaling & Behavior
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Autophagy has recently been shown to be required for postmeiotic anther development including anther dehiscence, programmed cell death-mediated degradation of the tapetum and pollen maturation in rice. Several phytohormones are known to play essential roles during male reproductive development including pollen maturation. However, the relationship between phytohormone metabolism and autophagy in plant reproductive development is unknown. We here comprehensively analyzed the effect of autophagy disruption on phytohormone contents in rice anthers at the flowering stage, and found that endogenous levels of active-forms of gibberellins (GAs) and cytokinin, trans-zeatin, were significantly lower in the autophagy-defective mutant, Osatg7–1, than in the wild type. Treatment with GA4 partially recovered maturation of the mutant pollens, but did not recover the limited anther dehiscence as well as sterility phenotype. These results suggest that autophagy affects metabolism and endogenous levels of GAs and cytokinin in rice anthers. Reduction in bioactive GAs in the autophagy-deficient mutant may partially explain the defects in pollen maturation of the autophagy-deficient mutant, but tapetal autophagy also plays other specific roles in fertilization.

ACS Style

Takamitsu Kurusu; Tomoko Koyano; Nobutaka Kitahata; Mikiko Kojima; Shigeru Hanamata; Hitoshi Sakakibara; Kazuyuki Kuchitsu. Autophagy-mediated regulation of phytohormone metabolism during rice anther development. Plant Signaling & Behavior 2017, 12, e1365211 .

AMA Style

Takamitsu Kurusu, Tomoko Koyano, Nobutaka Kitahata, Mikiko Kojima, Shigeru Hanamata, Hitoshi Sakakibara, Kazuyuki Kuchitsu. Autophagy-mediated regulation of phytohormone metabolism during rice anther development. Plant Signaling & Behavior. 2017; 12 (9):e1365211.

Chicago/Turabian Style

Takamitsu Kurusu; Tomoko Koyano; Nobutaka Kitahata; Mikiko Kojima; Shigeru Hanamata; Hitoshi Sakakibara; Kazuyuki Kuchitsu. 2017. "Autophagy-mediated regulation of phytohormone metabolism during rice anther development." Plant Signaling & Behavior 12, no. 9: e1365211.

Journal article
Published: 03 July 2017 in Plant and Cell Physiology
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ACS Style

Alex A.R. Webb; Kazuyuki Kuchitsu; June Kwak; Zhen-Ming Pei; Hidetoshi Iida. Sensors Make Sense of Signaling. Plant and Cell Physiology 2017, 58, 1121 -1125.

AMA Style

Alex A.R. Webb, Kazuyuki Kuchitsu, June Kwak, Zhen-Ming Pei, Hidetoshi Iida. Sensors Make Sense of Signaling. Plant and Cell Physiology. 2017; 58 (7):1121-1125.

Chicago/Turabian Style

Alex A.R. Webb; Kazuyuki Kuchitsu; June Kwak; Zhen-Ming Pei; Hidetoshi Iida. 2017. "Sensors Make Sense of Signaling." Plant and Cell Physiology 58, no. 7: 1121-1125.

Original research article
Published: 21 June 2017 in Frontiers in Plant Science
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Plants use stomatal closure mediated by elicitors as the first step of the innate immune response to restrict the microbial entry. We present a comprehensive study of the effect of cryptogein and harpin, two elicitors from microbial pathogens of tobacco, on stomatal closure and guard cell signaling components in Arabidopsis thaliana, a model plant. Cryptogein as well as harpin induced stomatal closure, while elevating the levels of reactive oxygen species (ROS) and nitric oxide (NO) in the guard cells of A. thaliana. Kinetic studies with fluorescent dyes revealed that the rise in ROS levels preceded that of NO in guard cells, when treated with these two elicitors. The restriction of NO levels in guard cells, even by ROS modulators indicates the essentiality of ROS for NO production during elicitor-triggered stomatal closure. The signaling events during elicitor-induced stomatal closure appear to converge at NADPH oxidase and ROS production. Our results provide the first report on stomatal closure associated with rise in ROS/NO of guard cells by cryptogein and harpin in A. thaliana. Our results establish that A. thaliana can be used to study stomatal responses to the typical elicitors from microbial pathogens of other plants. The suitability of Arabidopsis opens up an excellent scope for further studies on signaling events leading to stomatal closure by microbial elicitors.

ACS Style

Gunja Gayatri; Srinivas Agurla; Kazuyuki Kuchitsu; Kondreddy Anil; Appa R. Podile; Agepati S. Raghavendra. Stomatal Closure and Rise in ROS/NO of Arabidopsis Guard Cells by Tobacco Microbial Elicitors: Cryptogein and Harpin. Frontiers in Plant Science 2017, 8, 1096 -1096.

AMA Style

Gunja Gayatri, Srinivas Agurla, Kazuyuki Kuchitsu, Kondreddy Anil, Appa R. Podile, Agepati S. Raghavendra. Stomatal Closure and Rise in ROS/NO of Arabidopsis Guard Cells by Tobacco Microbial Elicitors: Cryptogein and Harpin. Frontiers in Plant Science. 2017; 8 ():1096-1096.

Chicago/Turabian Style

Gunja Gayatri; Srinivas Agurla; Kazuyuki Kuchitsu; Kondreddy Anil; Appa R. Podile; Agepati S. Raghavendra. 2017. "Stomatal Closure and Rise in ROS/NO of Arabidopsis Guard Cells by Tobacco Microbial Elicitors: Cryptogein and Harpin." Frontiers in Plant Science 8, no. : 1096-1096.

Jpr symposium
Published: 10 April 2017 in Journal of Plant Research
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Katsuyuki T. Yamato; Kazuyuki Kuchitsu. “Fusion” in fertilization: interdisciplinary collaboration among plant and animal scientists. Journal of Plant Research 2017, 130, 419 -421.

AMA Style

Katsuyuki T. Yamato, Kazuyuki Kuchitsu. “Fusion” in fertilization: interdisciplinary collaboration among plant and animal scientists. Journal of Plant Research. 2017; 130 (3):419-421.

Chicago/Turabian Style

Katsuyuki T. Yamato; Kazuyuki Kuchitsu. 2017. "“Fusion” in fertilization: interdisciplinary collaboration among plant and animal scientists." Journal of Plant Research 130, no. 3: 419-421.

Jpr symposium
Published: 31 March 2017 in Journal of Plant Research
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Autophagy is one of the major cellular processes of recycling of proteins, metabolites and intracellular organelles, and plays crucial roles in the regulation of innate immunity, stress responses and programmed cell death (PCD) in many eukaryotes. It is also essential in development and sexual reproduction in many animals. In plants, although autophagy-deficient mutants of Arabidopsis thaliana show phenotypes in abiotic and biotic stress responses, their life cycle seems normal and thus little had been known until recently about the roles of autophagy in development and reproduction. Rice mutants defective in autophagy show sporophytic male sterility and immature pollens, indicating crucial roles of autophagy during pollen maturation. Enzymatic production of reactive oxygen species (ROS) by respiratory burst oxidase homologues (Rbohs) play multiple roles in regulating anther development, pollen tube elongation and fertilization. Significance of autophagy and ROS in the regulation of PCD of transient cells during plant sexual reproduction is discussed in comparison with animals.

ACS Style

Takamitsu Kurusu; Kazuyuki Kuchitsu. Autophagy, programmed cell death and reactive oxygen species in sexual reproduction in plants. Journal of Plant Research 2017, 130, 491 -499.

AMA Style

Takamitsu Kurusu, Kazuyuki Kuchitsu. Autophagy, programmed cell death and reactive oxygen species in sexual reproduction in plants. Journal of Plant Research. 2017; 130 (3):491-499.

Chicago/Turabian Style

Takamitsu Kurusu; Kazuyuki Kuchitsu. 2017. "Autophagy, programmed cell death and reactive oxygen species in sexual reproduction in plants." Journal of Plant Research 130, no. 3: 491-499.

Journal article
Published: 01 February 2017 in Proceedings of the National Academy of Sciences
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As sessile organisms, plants have to accommodate to rapid changes in their surrounding environment. Reactive oxygen species (ROS) act as signaling molecules to transduce biotic and abiotic stimuli into plant stress adaptations. It is established that a respiratory burst oxidase homolog B of Nicotiana benthamiana (NbRBOHB) produces ROS in response to microbe-associated molecular patterns to inhibit pathogen infection. Plant viruses are also known as causative agents of ROS induction in infected plants; however, the function of ROS in plant–virus interactions remains obscure. Here, we show that the replication of red clover necrotic mosaic virus (RCNMV), a plant positive-strand RNA [(+)RNA] virus, requires NbRBOHB-mediated ROS production. The RCNMV replication protein p27 plays a pivotal role in this process, redirecting the subcellular localization of NbRBOHB and a subgroup II calcium-dependent protein kinase of N. benthamiana (NbCDPKiso2) from the plasma membrane to the p27-containing intracellular aggregate structures. p27 also induces an intracellular ROS burst in an RBOH-dependent manner. NbCDPKiso2 was shown to be an activator of the p27-triggered ROS accumulations and to be required for RCNMV replication. Importantly, this RBOH-derived ROS is essential for robust viral RNA replication. The need for RBOH-derived ROS was demonstrated for the replication of another (+)RNA virus, brome mosaic virus, suggesting that this characteristic is true for plant (+)RNA viruses. Collectively, our findings revealed a hitherto unknown viral strategy whereby the host ROS-generating machinery is diverted for robust viral RNA replication.

ACS Style

Kiwamu Hyodo; Kenji Hashimoto; Kazuyuki Kuchitsu; Nobuhiro Suzuki; Tetsuro Okuno. Harnessing host ROS-generating machinery for the robust genome replication of a plant RNA virus. Proceedings of the National Academy of Sciences 2017, 114, E1282 -E1290.

AMA Style

Kiwamu Hyodo, Kenji Hashimoto, Kazuyuki Kuchitsu, Nobuhiro Suzuki, Tetsuro Okuno. Harnessing host ROS-generating machinery for the robust genome replication of a plant RNA virus. Proceedings of the National Academy of Sciences. 2017; 114 (7):E1282-E1290.

Chicago/Turabian Style

Kiwamu Hyodo; Kenji Hashimoto; Kazuyuki Kuchitsu; Nobuhiro Suzuki; Tetsuro Okuno. 2017. "Harnessing host ROS-generating machinery for the robust genome replication of a plant RNA virus." Proceedings of the National Academy of Sciences 114, no. 7: E1282-E1290.

Journal article
Published: 13 October 2016 in Plant, Cell & Environment
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How plant tissues control their water behaviours (phase and movement) under subfreezing temperatures through adaptative strategies (freezing behaviours) is important for their survival. However, the fine details of freezing behaviours in complex organs and their regulation mechanisms are poorly understood, and non-invasive visualization/analysis is required. The localization/density of unfrozen water in wintering Cornus florida flower buds at subfreezing temperatures was visualized with high-resolution magnetic resonance imaging (MRI). This allowed tissue-specific freezing behaviours to be determined. MRI images revealed that individual anthers and ovules remained stably supercooled to -14 to -21 °C or lower. The signal from other floral tissues decreased during cooling to -7 °C, which likely indicates their extracellular freezing. Microscopic observation and differential thermal analyses revealed that the abrupt breakdown of supercooled individual ovules and anthers resulted in their all-or-nothing type of injuries. The distribution of ice nucleation activity in flower buds determined using a test tube-based assay corroborated which tissues primarily froze. MRI is a powerful tool for non-invasively visualizing unfrozen tissues. Freezing events and/or dehydration events can be located by digital comparison of MRI images acquired at different temperatures. Only anthers and ovules preferentially remaining unfrozen are a novel freezing behaviour in flower buds. Physicochemical and biological mechanisms/implications are discussed.

ACS Style

Masaya Ishikawa; Hiroyuki Ide; Hideyuki Yamazaki; Hiroki Murakawa; Kazuyuki Kuchitsu; William Price; Yoji Arata. Freezing behaviours in winteringCornus floridaflower bud tissues revisited using MRI. Plant, Cell & Environment 2016, 39, 2663 -2675.

AMA Style

Masaya Ishikawa, Hiroyuki Ide, Hideyuki Yamazaki, Hiroki Murakawa, Kazuyuki Kuchitsu, William Price, Yoji Arata. Freezing behaviours in winteringCornus floridaflower bud tissues revisited using MRI. Plant, Cell & Environment. 2016; 39 (12):2663-2675.

Chicago/Turabian Style

Masaya Ishikawa; Hiroyuki Ide; Hideyuki Yamazaki; Hiroki Murakawa; Kazuyuki Kuchitsu; William Price; Yoji Arata. 2016. "Freezing behaviours in winteringCornus floridaflower bud tissues revisited using MRI." Plant, Cell & Environment 39, no. 12: 2663-2675.