Updated on 2024/07/18

写真a

 
KATSUHARA Maki
 
Organization
Institute of Plant Science and Resources Professor
Position
Professor
External link

Degree

  • Doctor of Science ( The University of Tokyo )

Research Interests

  • membrane transport

  • plant physiology

  • salt tolerance

  • stress physiology

  • 植物生理学

  • aquaporin

  • ストレス生理学

  • 膜輸送

  • アクアポリン

  • 耐塩性

Research Areas

  • Life Science / Plant nutrition and soil science

  • Life Science / Plant molecular biology and physiology

Education

  • The University of Tokyo   大学院理学系研究科植物学専攻博士課程  

    1987.4 - 1990.3

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    Country: Japan

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  • The University of Tokyo   大学院理学系研究科植物学専攻修士課程  

    1985.4 - 1987.3

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    Country: Japan

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  • The University of Tokyo   理学部   生物学教室

    1981.4 - 1985.3

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    Country: Japan

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Research History

  • Okayama University   Institute of Plant Science and Resources   Professor

    2017.4

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    Country:Japan

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  • Okayama University   Institute of Plant Science and Resources   Associate Professor

    2003.10 - 2017.3

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    Country:Japan

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  • Okayama University   Research Institute for Bioresources   Assistant Professor

    1992.4 - 2003.9

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    Country:Japan

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  • Insect Agricultural Technology Research Institute   Research Fellow

    1990.10 - 1992.3

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    Country:Japan

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  • The University of Tokyo   Faculty of Science   Post-doctoral researcher

    1990.4 - 1990.9

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    Country:Japan

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Professional Memberships

Committee Memberships

  • The Japanese Society of Plant Physiologists   Representatives  

    2022.1 - 2023.12   

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    Committee type:Academic society

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  • 日本植物学会   編集担当理事  

    2021.3   

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  • 鳥取大学乾燥地研究センター   共同研究委員(外部委員)  

    2020.4   

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    Committee type:Other

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  • 根研究学会   評議員  

    2020.4   

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  • 日本植物学会   編集実行委員  

    2012 - 2016   

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    Committee type:Academic society

    日本植物学会

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  • 日本植物生理学会   代議員  

    2012 - 2014   

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    Committee type:Academic society

    日本植物生理学会

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  • 根研究会   英文誌編集委員会委員  

    2007   

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    根研究会

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Papers

  • In planta evidence that the HAK transporter OsHAK2 is involved in Na+ transport in rice. International journal

    Shigeto Morita, Natsumi Tamba, Mineo Shibasaka, Shizuka Sasano, Taiju Kadoike, Yasuyo Urase, Masamitsu Maruyama, Aguri Fukuoka, Junta Yanai, Takehiro Masumura, Yasunari Ogihara, Shigeru Satoh, Kunisuke Tanaka, Maki Katsuhara, Hideki Nakayama

    Bioscience, biotechnology, and biochemistry   87 ( 5 )   482 - 490   2023.4

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    HAK family transporters primarily function as K+ transporters and play major roles in K+ uptake and translocation in plants, whereas several HAK transporters exhibit Na+ transport activity. OsHAK2, a rice HAK transporter, was shown to mediate Na+ transport in Escherichia coli in a previous study. In this study, we investigated whether OsHAK2 is involved in Na+ transport in the rice plant. Overexpression of OsHAK2 increased Na+ translocation from the roots to the shoots of transgenic rice. It also increased both root and whole-plant Na+ content, and enhanced shoot length under low Na+ and K+ conditions. Meanwhile, OsHAK2 overexpression increased salt sensitivity under a long-term salt stress condition, indicating that OsHAK2 is not involved in salt tolerance, unlike in the case of ZmHAK4 in maize. These results suggest that OsHAK2 is permeable to Na+ and contributes to shoot growth in rice plants under low Na+ and K+ conditions.

    DOI: 10.1093/bbb/zbad020

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  • Distinct Functions of the Atypical Terminal Hydrophilic Domain of the HKT Transporter in the Liverwort Marchantia polymorpha. Reviewed International coauthorship International journal

    Shahin Imran, Masumi Oyama, Rie Horie, Natsuko I Kobayashi, Alex Costa, Ryosuke Kumano, Chiho Hirata, Sen Thi Huong Tran, Maki Katsuhara, Keitaro Tanoi, Takayuki Kohchi, Kimitsune Ishizaki, Tomoaki Horie

    Plant & Cell Physiology   63 ( 6 )   802 - 816   2022.6

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    K+/Na+ homeostasis is important for land plants, particularly under salt stress. In this study, the structure and ion transport properties of the high-affinity K+ transporter (HKT) of the liverwort Marchantia polymorpha were investigated. Only one HKT gene, MpHKT1, was identified in the genome of M. polymorpha. Phylogenetic analysis of HKT proteins revealed that non-seed plants possess HKTs grouped into a clade independent of the other two clades including HKTs of angiosperms. A distinct long hydrophilic domain was found in the C-terminus of MpHKT1. Complementary DNA (cDNA) of truncated MpHKT1 (t-MpHKT1) encoding the MpHKT_Δ596-812 protein was used to examine the functions of the C-terminal domain. Both MpHKT1 transporters fused with enhanced green fluorescent protein at the N-terminus were localized to the plasma membrane when expressed in rice protoplasts. Two-electrode voltage clamp experiments using Xenopus laevis oocytes indicated that MpHKT1 mediated the transport of monovalent alkali cations with higher selectivity for Na+ and K+, but truncation of the C-terminal domain significantly reduced the transport activity with a decrease in the Na+ permeability. Overexpression of MpHKT1 or t-MpHKT1 in M. polymorpha conferred accumulation of higher Na+ levels and showed higher Na+ uptake rates, compared to those of wild-type plants; however, phenotypes with t-MpHKT1 were consistently weaker than those with MpHKT1. Together, these findings suggest that the hydrophilic C-terminal domain plays a unique role in the regulation of transport activity and ion selectivity of MpHKT1.

    DOI: 10.1093/pcp/pcac044

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  • Calcium control of the hydraulic resistance in cells of Chara corallina Reviewed International coauthorship International journal

    Masashi Tazawa, Maki Katsuhara, Randy Wayne

    Protoplasma   260 ( 1 )   299 - 306   2022.6

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media LLC  

    DOI: 10.1007/s00709-022-01772-z

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    Other Link: https://link.springer.com/article/10.1007/s00709-022-01772-z/fulltext.html

  • Identification and Characterization of Rice OsHKT1;3 Variants Reviewed International journal

    Shahin Imran, Yoshiyuki Tsuchiya, Sen Thi Huong Tran, Maki Katsuhara

    Plants   10 ( 10 )   2006 - 2006   2021.9

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    Authorship:Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:MDPI AG  

    In rice, the high-affinity K+ transporter, OsHKT1;3, functions as a Na+-selective transporter. mRNA variants of OsHKT1;3 have been reported previously, but their functions remain unknown. In this study, five OsHKT1;3 variants (V1-V5) were identified from japonica rice (Nipponbare) in addition to OsHKT1;3_FL. Absolute quantification qPCR analyses revealed that the transcript level of OsHKT1;3_FL was significantly higher than other variants in both the roots and shoots. Expression levels of OsHKT1;3_FL, and some variants, increased after 24 h of salt stress. Two electrode voltage clamp experiments in a heterologous expression system using Xenopus laevis oocytes revealed that oocytes expressing OsHKT1;3_FL and all of its variants exhibited smaller Na+ currents. The presented data, together with previous data, provide insights to understanding how OsHKT family members are involved in the mechanisms of ion homeostasis and salt tolerance in rice.

    DOI: 10.3390/plants10102006

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  • Physiological Role of Aerobic Fermentation Constitutively Expressed in an Aluminum-Tolerant Cell Line of Tobacco (Nicotiana tabacum) Reviewed International journal

    Tsuchiya Y, Nakamura T, Izumi Y, Okazaki K, Shinano T, Kubo Y, Katsuhara M, Sasaki T, Yamamoto Y

    Plant & Cell Physiology   62 ( 9 )   1460 - 1477   2021.9

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Oxford University Press (OUP)  

    <title>Abstract</title>
    Aluminum (Al)-tolerant tobacco cell line ALT301 derived from SL (wild-type) hardly exhibits Al-triggered reactive oxygen species (ROS) compared with SL. Molecular mechanism leading to this phenotype was investigated comparatively with SL. Under normal growth condition, metabolome data suggested the activation of glycolysis and lactate fermentation but the repression of the tricarboxylic acid (TCA) cycle in ALT301, namely aerobic fermentation, which seemed to be transcriptionally controlled partly by higher expression of genes encoding lactate dehydrogenase and pyruvate dehydrogenase kinase. Microarray and gene ontology analyses revealed the upregulation of th e gene encoding related to APETALA2.3 (RAP2.3)-like protein, one of the group VII ethylene response factors (ERFVIIs), in ALT301. ERFVII transcription factors are known to be key regulators for hypoxia response that promotes substrate-level ATP production by glycolysis and fermentation. ERFVIIs are degraded under normoxia by the N-end rule pathway of proteolysis depending on both oxygen and nitric oxide (NO), and NO is produced mainly by nitrate reductase (NR) in plants. In ALT301, levels of the NR gene expression (NIA2), NR activity and NO production were all lower compared with SL. Consistently, the known effects of NO on respiratory pathways were also repressed in ALT301. Under Al-treatment condition, NO level increased in both lines but was lower in ALT301. These results suggest that the upregulation of the RAP2.3-like gene and the downregulation of the NIA2 gene and resultant NO depletion in ALT301 coordinately enhance aerobic fermentation, which seems to be related to a higher capacity to prevent ROS production in mitochondria under Al stress.

    DOI: 10.1093/pcp/pcab098

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  • Age dependence of the hydraulic resistances of the plasma membrane and the tonoplast (vacuolar membrane) in cells of Chara corallina Reviewed International coauthorship International journal

    Masashi Tazawa, Maki Katsuhara, Randy Wayne

    Protoplasma   258 ( 4 )   793 - 801   2021.7

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media LLC  

    DOI: 10.1007/s00709-020-01596-9

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    Other Link: https://link.springer.com/article/10.1007/s00709-020-01596-9/fulltext.html

  • Erratum: Physiological role of aerobic fermentation constitutively expressed in an aluminum-tolerant cell line of tobacco (Nicotiana Tabacum) (Plant and Cell Physiology DOI: 10.1093/pcp/pcab098)

    Yoshiyuki Tsuchiya, Takuji Nakamura, Yohei Izumi, Keiki Okazaki, Takuro Shinano, Yasutaka Kubo, Maki Katsuhara, Takayuki Sasaki, Yoko Yamamoto

    Plant and Cell Physiology   62 ( 6 )   1058   2021.6

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Oxford University Press  

    In the originally published version of this manuscript, the ethylene signaling pathway in Arabidopsis shown in the Figure 3 was incorrectly referred. The correct pathway is as follows: Ethylene -| ETR1 -&gt
    CTR1 -| EIN2 -&gt
    EIN3/EIL1 -&gt
    ERFs. In addition, information was missing from the Figure 3 caption. An explanation of the subgroups of the ERF family shown in the figure (Group IX, VIII, VI, VII) has been added to the caption, which also necessitated two new references in the paper. The corrected caption is: Expression of the genes encoding transcription factors (EIL, ERFs) and chitinases (CHNs) associated with ethylene signaling pathway in SL and ALT301 under normal growth condition. Gene expression analysis was performed by real-time RT-PCR. Values are the mean±SE (n=3–5 from three to five independent experiments, respectively). Significant differences between SL and ALT301 are indicated with asterisks (*P &lt
    0.05, **P &lt
    0.01, Welch’s t-test). Ethylene signaling pathway in Arabidopsis (Kazan 2015) is shown (top). Group names of the ERF genes are indicated, which is based on Rushton et al. (2008) and Nakano et al. (2006). Note that some of the ERF genes have several synonyms (see the National Center for Biotechnology Information [NCBI] database using the gene accessions listed in Supplementary Table S6). Abbreviations: ETR1, Ethylene Response1
    CTR1, Constitutive Triple Response1
    EIN2, Ethylene Insensitive2
    EIN3, Ethylene Insensitive3
    EIL1, Ethylene Insensitive-Like Protein1
    ERF, Ethylene Response Factor. The gene Tsi1 encodes Tobacco stress-induced gene1. The two new references are: Nakano, T., Suzuki, K., Fujimura, T. and Shinshi, H. (2006) Genome-wide analysis of the ERF gene family in Arabidopsis and rice. Plant Physiol. 140: 411–432. Rushton, P.J., Bokowiec, M.T., Han, S., Zhang, H., Brannock, J.F., Chen, X., et al. (2008) Tobacco transcription factors: novel insights into transcriptional regulation in the Solanaceae. Plant Physiol. 147: 280–295. These errors have been corrected.

    DOI: 10.1093/pcp/pcab131

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  • Mechanisms Activating Latent Functions of PIP Aquaporin Water Channels via the Interaction between PIP1 and PIP2 Proteins Reviewed International journal

    Mineo Shibasaka, Tomoaki Horie, Maki Katsuhara

    Plant & Cell Physiology   62 ( 1 )   92 - 99   2021.3

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    Authorship:Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Oxford University Press (OUP)  

    <title>Abstract</title>
    Plant plasma membrane-type plasma membrane intrinsic protein (PIP) aquaporins are classified into two groups, PIP1s and PIP2s. In this study, we focused on HvPIP1;2, a PIP1 in barley (Hordeum vulgare), to dissect the molecular mechanisms that evoke HvPIP1-mediated water transport. No HvPIP1;2 protein was localized to the plasma membrane when expressed alone in Xenopus laevis oocytes. By contrast, a chimeric HvPIP1;2 protein (HvPIP1;2_24NC), in which the N- and C-terminal regions were replaced with the corresponding regions from HvPIP2;4, was found to localize to the plasma membrane of oocytes. However, HvPIP1;2_24NC showed no water transport activity in swelling assays. These results suggested that the terminal regions of PIP2 proteins direct PIP proteins to the plasma membrane, but the relocalization of PIP1 proteins was not sufficient to PIP1s functionality as a water channel in a membrane. A single amino acid replacement of threonine by methionine in HvPIP2;4 (HvPIP2;4T229M) abolished water transport activity. Co-expression of HvPIP1;2_24NC either with HvPIP2;4_12NC or with HvPIP2;4TM_12NC, in which the N- and C-terminal regions were replaced with the corresponding regions of HvPIP1;2, increased the water transport activity in oocytes. These data provided evidence that the HvPIP1;2 molecule has own water transport activity and an interaction with the middle part of the HvPIP2;4 protein (except for the N- and C-termini) is required for HvPIP1;2 functionality as a water channel. This molecular mechanism could be applied to other PIP1s and PIP2s in addition to the known mechanism that the terminal regions of some PIP2s lead some PIP1s to the plasma membrane.

    DOI: 10.1093/pcp/pcaa142

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    Other Link: http://academic.oup.com/pcp/article-pdf/62/1/92/36718180/pcaa142.pdf

  • Accession difference in leaf photosynthesis, root hydraulic conductance and gene expression of root aquaporins under salt stress in barley seedlings Reviewed International coauthorship International journal

    Asuka Kodama, Tamaki Watanabe, Makoto Yamaguchi, Ryohei Narita, Maki Katsuhara, Kazuhiro Sato, Taiichiro Ookawa, Tadashi Hirasawa

    Plant Production Science   24 ( 1 )   73 - 82   2021.1

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    DOI: 10.1080/1343943x.2020.1794915

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  • A Survey of Barley PIP Aquaporin Ionic Conductance Reveals Ca2+-Sensitive HvPIP2;8 Na+ and K+ Conductance Reviewed International coauthorship International journal

    Sen Thi Huong Tran, Tomoaki Horie, Shahin Imran, Jiaen Qiu, Samantha McGaughey, Caitlin S. Byrt, Stephen D. Tyerman, Maki Katsuhara

    International Journal of Molecular Sciences   21 ( 19 )   7135 - 7135   2020.9

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    Some plasma membrane intrinsic protein (PIP) aquaporins can facilitate ion transport. Here we report that one of the 12 barley PIPs (PIP1 and PIP2) tested, HvPIP2;8, facilitated cation transport when expressed in Xenopus laevis oocytes. HvPIP2;8-associated ion currents were detected with Na+ and K+, but not Cs+, Rb+, or Li+, and was inhibited by Ba2+, Ca2+, and Cd2+ and to a lesser extent Mg2+, which also interacted with Ca2+. Currents were reduced in the presence of K+, Cs+, Rb+, or Li+ relative to Na+ alone. Five HvPIP1 isoforms co-expressed with HvPIP2;8 inhibited the ion conductance relative to HvPIP2;8 alone but HvPIP1;3 and HvPIP1;4 with HvPIP2;8 maintained the ion conductance at a lower level. HvPIP2;8 water permeability was similar to that of a C-terminal phosphorylation mimic mutant HvPIP2;8 S285D, but HvPIP2;8 S285D showed a negative linear correlation between water permeability and ion conductance that was modified by a kinase inhibitor treatment. HvPIP2;8 transcript abundance increased in barley shoot tissues following salt treatments in a salt-tolerant cultivar Haruna-Nijo, but not in salt-sensitive I743. There is potential for HvPIP2;8 to be involved in barley salt-stress responses, and HvPIP2;8 could facilitate both water and Na+/K+ transport activity, depending on the phosphorylation status.

    DOI: 10.3390/ijms21197135

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  • Na+ Transporter SvHKT1;1 from a Halophytic Turf Grass Is Specifically Upregulated by High Na+ Concentration and Regulates Shoot Na+ Concentration Reviewed International journal

    Yuki Kawakami, Shahin Imran, Maki Katsuhara, Yuichi Tada

    International Journal of Molecular Sciences   21 ( 17 )   6100 - 6100   2020.8

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    We characterized an Na+ transporter SvHKT1;1 from a halophytic turf grass, Sporobolus virginicus. SvHKT1;1 mediated inward and outward Na+ transport in Xenopus laevis oocytes and did not complement K+ transporter-defective mutant yeast. SvHKT1;1 did not complement athkt1;1 mutant Arabidopsis, suggesting its distinguishable function from other typical HKT1 transporters. The transcript was abundant in the shoots compared with the roots in S. virginicus and was upregulated by severe salt stress (500 mM NaCl), but not by lower stress. SvHKT1;1-expressing Arabidopsis lines showed higher shoot Na+ concentrations and lower salt tolerance than wild type (WT) plants under nonstress and salt stress conditions and showed higher Na+ uptake rate in roots at the early stage of salt treatment. These results suggested that constitutive expression of SvHKT1;1 enhanced Na+ uptake in root epidermal cells, followed by increased Na+ transport to shoots, which led to reduced salt tolerance. However, Na+ concentrations in phloem sap of the SvHKT1;1 lines were higher than those in WT plants under salt stress. Based on this result, together with the induction of the SvHKT1;1 transcription under high salinity stress, it was suggested that SvHKT1;1 plays a role in preventing excess shoot Na+ accumulation in S. virginicus.

    DOI: 10.3390/ijms21176100

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  • Ectopic expression of a rice plasma membrane intrinsic protein (OsPIP1;3) promotes plant growth and water uptake Reviewed International coauthorship International journal

    Siyu Liu, Tatsuya Fukumoto, Patrizia Gena, Peng Feng, Qi Sun, Qiang Li, Tadashi Matsumoto, Toshiyuki Kaneko, Hang Zhang, Yao Zhang, Shihua Zhong, Weizhong Zeng, Maki Katsuhara, Yoshichika Kitagawa, Aoxue Wang, Giuseppe Calamita, Xiaodong Ding

    The Plant Journal   102 ( 4 )   779 - 796   2020.5

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Wiley  

    DOI: 10.1111/tpj.14662

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    Other Link: https://onlinelibrary.wiley.com/doi/full-xml/10.1111/tpj.14662

  • Expression and Ion Transport Activity of Rice OsHKT1;1 Variants Reviewed International journal

    Shahin Imran, Tomoaki Horie, Maki Katsuhara

    Plants   9 ( 1 )   16 - 16   2019.12

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    OsHKT1;1 in rice, belongs to the high-affinity K+ Transporter family, has been found to be involved in salt tolerance. OsHKT1;1 in japonica rice (Nipponbare) produces mRNA variants, but their functions remain elusive. In salt tolerant rice, Pokkali, eight OsHKT1;1 variants (V1-V8) were identified in addition to the full-length OsHKT1;1 (FL) cDNA. Absolute quantification by qPCR revealed that accumulation of OsHKT1;1-FL mRNA is minor in contrast to that of OsHKT1;1-V1, -V2, -V4, and -V7 mRNAs, all of which are predominant in shoots, while only V1 and V7 mRNAs are predominant in roots. Two electrode voltage clamp (TEVC) experiments using Xenopus laevis oocytes revealed that oocytes-expressing OsHKT1;1-FL from Pokkali exhibited inward-rectified currents in the presence of 96 mM Na+ as reported previously. Further TEVC analyses indicated that six of eight OsHKT1;1 variants elicited currents in a Na+ or a K+ bath solution. OsHKT1;1-V6 exhibited a similar inward rectification to the FL protein. Contrastingly, however, the rests mediated bidirectional currents in both Na+ and K+ bath solutions. These data suggest possibilities that novel mechanisms regulating the transport activity of OsHKT1;1 might exist, and that OsHKT1;1 variants might also carry out distinct physiological roles either independently or in combination with OsHKT1;1-FL.

    DOI: 10.3390/plants9010016

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  • Dynamics of the contents and distribution of ABA, auxins and aquaporins in developing caryopses of an ABA-deficient barley mutant and its parental cultivar Reviewed

    Oksana A. Seldimirova, Guzel R. Kudoyarova, Maki Katsuhara, Ilshat R. Galin, Denis Yu. Zaitsev, Natalia N. Kruglova, Dmitry S. Veselov, Stanislav Yu. Veselov

    Seed Science Research   29 ( 4 )   261 - 269   2019.12

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Cambridge University Press (CUP)  

    <title>Abstract</title>Dynamics of abscisic acid (ABA) and indole-3-acetic acid (IAA) contents were followed in developing barley caryopses of the ABA-deficient mutant AZ34 and its parental cultivar Steptoe. Distribution of these hormones and HvPIP2 aquaporins (AQPs) was studied with the help of immunohistochemical methods in the roots and coleorhiza of developing embryos. In Steptoe, maturation of the caryopsis was accompanied by vast accumulation of ABA, while this hormone accumulated more slowly in the caryopsis of AZ34 and its content was lower than in Steptoe. Accumulation of ABA was accompanied by a decline in IAA level in the developing caryopsis, the process being delayed in AZ34 in accordance with the slower accumulation of ABA. ABA accumulated to high levels in the coleorhiza cells of Steptoe, while the effect was absent in AZ34. The high level of ABA was likely to be important for maintaining the barrier function of the coleorhiza, preventing germination of seminal roots and enabling seed dormancy, while the absence of ABA accumulation in coleorhiza of AZ34 may be responsible for the initiation of root germination inside the caryopsis. The abundance of HvPIP2 AQPs in the seminal roots was higher at the beginning of maturation of Steptoe caryopsis and declined afterwards, while the levels of APQs increased later in AZ34 in accordance with the delay in ABA accumulation. These results suggest the importance of ABA accumulation in coleorhiza for preventing precocious growth of seminal roots, and suggest regulation of IAA and aquaporin levels by this hormone during maturation of embryos.

    DOI: 10.1017/s0960258519000229

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  • The Mechanism of SO2 -Induced Stomatal Closure Differs from O3 and CO2 Responses and Is Mediated by Non-Apoptotic Cell Death in Guard Cells. Reviewed International journal

    Ooi L, Matsuura T, Munemasa S, Murata Y, Katsuhara M, Hirayama T, Mori IC

    Plant, cell & environment   42 ( 2 )   437 - 447   2019.2

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    Plants closing stomata in the presence of harmful gases is believed to be a stress avoidance mechanism. SO2 , one of the major airborne pollutants, has long been reported to induce stomatal closure, yet the mechanism remains unknown. Little is known about the stomatal response to airborne pollutants besides O3 . SLOW ANION CHANNEL-ASSOCIATED 1 (SLAC1) and OPEN STOMATA 1 (OST1) were identified as genes mediating O3 -induced closure. SLAC1 and OST1 are also known to mediate stomatal closure in response to CO2 , together with RESPIRATORY BURST OXIDASE HOMOLOGs (RBOHs). The overlaying roles of these genes in response to O3 and CO2 suggested that plants share their molecular regulators for airborne stimuli. Here, we investigated and compared stomatal closure event induced by a wide concentration range of SO2 in Arabidopsis through molecular genetic approaches. O3 - and CO2 -insensitive stomata mutants did not show significant differences from the wild type in stomatal sensitivity, guard cell viability, and chlorophyll content revealing that SO2 -induced closure is not regulated by the same molecular mechanisms as for O3 and CO2 . Nonapoptotic cell death is shown as the reason for SO2 -induced closure, which proposed the closure as a physicochemical process resulted from SO2 distress, instead of a biological protection mechanism.

    DOI: 10.1111/pce.13406

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  • High-affinity K+ transporters from a halophyte, Sporobolus virginicus, mediate both K+ and Na+ transport in transgenic Arabidopsis, X. laevis oocytes, and yeast. Reviewed

    Tada Y, Endo C, Katsuhara M, Horie T, Shibasaka M, Nakahara Y, Kurusu T

    Plant & Cell Physiology   60 ( 1 )   176 - 187   2019.1

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    DOI: 10.1093/pcp/pcy202

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  • Functional screening of salt tolerance genes from a halophyte Sporobolus virginicus and transcriptomic and metabolomic analysis of salt tolerant plants expressing glycine-rich RNA-binding protein Reviewed

    Yuichi Tada, Ryuichi Kawano, Shiho Komatsubara, Hideki Nishimura, Maki Katsuhara, Soichi Ozaki, Shin Terashima, Kentaro Yano, Chisato Endo, Muneo Sato, Mami Okamoto, Yuji Sawada, Masami Yokota Hirai, Takamitsu Kurusu

    Plant Science   278   54 - 63   2019.1

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    DOI: 10.1016/j.plantsci.2018.10.019

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  • Energy costs of salinity tolerance in crop plants Invited Reviewed

    Stephen D. Tyerman, Rana Munns, Wieland Fricke, Borjana Arsova, Bronwyn J. Barkla, Jayakumar Bose, Helen Bramley, Caitlin Byrt, Zhonghua Chen, Timothy D. Colmer, Tracey Cuin, David A. Day, Kylie J. Foster, Matthew Gilliham, Sam W. Henderson, Tomoaki Horie, Colin L. D. Jenkins, Brent N. Kaiser, Maki Katsuhara, Darren Plett, Stanley J. Miklavcic, Stuart J. Roy, Francisco Rubio, Sergey Shabala, Megan Shelden, Kathleen Soole, Nicolas L. Taylor, Mark Tester, Michelle Watt, Stefanie Wege, Lars H. Wegner, Zhengyu Wen

    New Phytologist   221 ( 1 )   25 - 29   2019.1

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Wiley  

    DOI: 10.1111/nph.15555

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  • A Cyclic Nucleotide-Gated Channel, HvCNGC2-3, Is Activated by the Co-Presence of Na+ and K+ and Permeable to Na+ and K+ Non-Selectively Reviewed International journal

    Izumi Mori, Yuichi Nobukiyo, Yoshiki Nakahara, Mineo Shibasaka, Takuya Furuichi, Maki Katsuhara

    Plants   7 ( 3 )   61 - 61   2018.7

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    Cyclic nucleotide-gated channels (CNGCs) have been postulated to contribute significantly in plant development and stress resistance. However, their electrophysiological properties remain poorly understood. Here, we characterized barley CNGC2-3 (HvCNGC2-3) by the two-electrode voltage-clamp technique in the Xenopus laevis oocyte heterologous expression system. Current was not observed in X. laevis oocytes injected with HvCNGC2-3 complementary RNA (cRNA) in a bathing solution containing either Na+ or K+ solely, even in the presence of 8-bromoadenosine 3′,5′-cyclic monophosphate (8Br-cAMP) or 8-bromoguanosine 3′,5′-cyclic monophosphate (8Br-cGMP). A weakly voltage-dependent slow hyperpolarization-activated ion current was observed in the co-presence of Na+ and K+ in the bathing solution and in the presence of 10 µM 8Br-cAMP, but not 8Br-cGMP. Permeability ratios of HvCNGC2-3 to K+, Na+ and Cl− were determined as 1:0.63:0.03 according to reversal-potential analyses. Amino-acid replacement of the unique ion-selective motif of HvCNGC2-3, AQGL, with the canonical motif, GQGL, resulted in the abolition of the current. This study reports a unique two-ion-dependent activation characteristic of the barley CNGC, HvCNGC2-3.

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  • T-DNA Tagging-Based Gain-of-Function of OsHKT1;4 Reinforces Na Exclusion from Leaves and Stems but Triggers Na Toxicity in Roots of Rice Under Salt Stress Reviewed

    Oda Y, Kobayashi NI, Tanoi K, Ma JF, Itou Y, Katsuhara M, Itou T, Horie T

    International Journal of Molecular Sciences   19 ( 1 )   2018.1

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    The high affinity K+ transporter 1;4 (HKT1;4) in rice (Oryza sativa), which shows Na+ selective transport with little K+ transport activity, has been suggested to be involved in reducing Na in leaves and stems under salt stress. However, detailed physiological roles of OsHKT1;4 remain unknown. Here, we have characterized a transfer DNA (T-DNA) insertion mutant line of rice, which overexpresses OsHKT1;4, owing to enhancer elements in the T-DNA, to gain an insight into the impact of OsHKT1;4 on salt tolerance of rice. The homozygous mutant (the O/E line) accumulated significantly lower concentrations of Na in young leaves, stems, and seeds than the sibling WT line under salt stress. Interestingly, however, the mutation rendered the O/E plants more salt sensitive than WT plants. Together with the evaluation of biomass of rice lines, rhizosphere acidification assays using a pH indicator bromocresol purple and (NaCl)-Na-22 tracer experiments have led to an assumption that roots of O/E plants suffered heavier damages from Na which excessively accumulated in the root due to increased activity of Na+ uptake and Na+ exclusion in the vasculature. Implications toward the application of the HKT1-mediated Na+ exclusion system to the breeding of salt tolerant crop cultivars will be discussed.

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    Other Link: http://orcid.org/0000-0001-7545-2771

  • Identification of an H2O2permeable PIP aquaporin in barley and a serine residue promoting H2O2transport Reviewed

    Rhee J, Horie T, Sasano S, Nakahara Y, Katsuhara M

    Physiologia Plantarum   159 ( 1 )   120 - 128   2017.1

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  • Exogenous application of abscisic acid (ABA) increases root and cell hydraulic conductivity and abundance of some aquaporin isoforms in the ABA-deficient barley mutant Az34 Reviewed

    Guzel Sharipova, Dmitriy Veselov, Guzel Kudoyarova, Wieland Fricke, Ian C. Dodd, Maki Katsuhara, Takuya Furuichi, Igor Ivanov, Stanislav Veselov

    Annals of Botany   118 ( 4 )   777 - 785   2016.10

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  • Genome-Wide Characterization of Major Intrinsic Proteins in Four Grass Plants and Their Non-Aqua Transport Selectivity Profiles with Comparative Perspective

    Abul Kalam Azad, Jahed Ahmed, Md. Asraful Alum, Md. Mahbub Hasan, Takahiro Ishikawa, Yoshihiro Sawa, Maki Katsuhara

    PLOS ONE   11 ( 6 )   e0157735   2016.6

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    Major intrinsic proteins (MIPs), commonly known as aquaporins, transport not only water in plants but also other substrates of physiological significance and heavy metals. In most of the higher plants, MIPs are divided into five subfamilies (PIPs, TIPs, NIPs, SIPs and XIPs). Herein, we identified 68, 42, 38 and 28 full-length MIPs, respectively in the genomes of four monocot grass plants, specifically Panicum virgatum, Setaria italica, Sorghum bicolor and Brachypodium distachyon. Phylogenetic analysis showed that the grass plants had only four MIP subfamilies including PIPs, TIPs, NIPs and SIPs without XIPs. Based on structural analysis of the homology models and comparing the primary selectivity-related motifs [two NPA regions, aromatic/arginine (ar/R) selectivity filter and Froger's positions (FPs)] of all plant MIPs that have been experimentally proven to transport non-aqua substrates, we predicted the transport profiles of all MIPs in the four grass plants and also in eight other plants. Groups of MIP subfamilies based on ar/R selectivity filter and FPs were linked to the non-aqua transport profiles. We further deciphered the substrate selectivity profiles of the MIPs in the four grass plants and compared them with their counterparts in rice, maize, soybean, poplar, cotton, Arabidopsis thaliana, Physcomitrella patens and Selaginella moellendorffii. In addition to two NPA regions, ar/R filter and FPs, certain residues, especially in loops B and C, contribute to the functional distinctiveness of MIP groups. Expression analysis of transcripts in different organs indicated that non-aqua transport was related to expression of MIPs since most of the unexpressed MIPs were not predicted to facilitate the transport of non-aqua molecules. Among all MIPs in every plant, TIP (BdTIP1;1, SiTIP1;2, SbTIP2;1 and PvTIP1;2) had the overall highest mean expression. Our study generates significant information for understanding the diversity, evolution, non-aqua transport profiles and insight into comparative transport selectivity of plant MIPs, and provides tools for the development of transgenic plants.

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  • Phosphorus deprivation effects on water relations of Nicotiana tabacum plant via reducing plasma membrane permeability

    M. Mahdieh, A. Mostajeran, M. Katsuhara

    RUSSIAN JOURNAL OF PLANT PHYSIOLOGY   63 ( 1 )   54 - 61   2016.1

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    Plants grown in phosphorus-deprived solutions often exhibit disruption of water transport due to reduction in root hydraulic conductivity (Lp (r) ). To uncover the relationship between root Lp (r) and water permeability coefficient (P (f)) of plasma membrane and the role of aquaporins, we evaluated P (f) of plasma membrane and also PIP-type aquaporin gene expression in tobacco (Nicotiana tabacum L.) plant roots after seven days P-deprivation. The results showed significant reduction in sap flow rate (J (v)) and osmotic root hydraulic conductivity (L (pr-o)) in P-deprived roots. These effects were reversed 24 h after P-resupplying. Interestingly, the P (f) of root protoplasts was 57% lower in P-deprived plants compared with P-sufficient ones. The expression of NtPIP1;1 and NtPIP2;1 aquaporins did not change significantly in P-deprived plants compared with P-sufficient ones, but the copy number of NtAQP1 increased significantly in P-deprived plants. P-deprivation did not change Lp r-o significantly in antisense NtAQP1 plants. Taken together, these findings suggest that P-deprivation may play an important role in modulation of root hydraulic conductivity by affecting P (f) in transcellular pathway of water flow across roots and aquaporins. Finally, we concluded that dominant water transport pathway under P-deprivation was transcellular one.

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  • OsHKT1;4-mediated Na+ transport in stems contributes to Na+ exclusion from leaf blades of rice at the reproductive growth stage upon salt stress

    Kei Suzuki, Naoki Yamaji, Alex Costa, Eiji Okuma, Natsuko I. Kobayashi, Tatsuhiko Kashiwagi, Maki Katsuhara, Cun Wang, Keitaro Tanoi, Yoshiyuki Murata, Julian I. Schroeder, Jian Feng Ma, Tomoaki Horie

    BMC PLANT BIOLOGY   16 ( 1 )   22   2016.1

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    Background: Na+ exclusion from leaf blades is one of the key mechanisms for glycophytes to cope with salinity stress. Certain class I transporters of the high-affinity K+ transporter (HKT) family have been demonstrated to mediate leaf blade-Na+ exclusion upon salinity stress via Na+-selective transport. Multiple HKT1 transporters are known to function in rice (Oryza sativa). However, the ion transport function of OsHKT1;4 and its contribution to the Na+ exclusion mechanism in rice remain to be elucidated.
    Results: Here, we report results of the functional characterization of the OsHKT1; 4 transporter in rice. OsHKT1; 4 mediated robust Na+ transport in Saccharomyces cerevisiae and Xenopus laevis oocytes. Electrophysiological experiments demonstrated that OsHKT1; 4 shows strong Na+ selectivity among cations tested, including Li+, Na+, K+, Rb+, Cs+, and NH4+, in oocytes. A chimeric protein, EGFP-OsHKT1;4, was found to be functional in oocytes and targeted to the plasma membrane of rice protoplasts. The level of OsHKT1; 4 transcripts was prominent in leaf sheaths throughout the growth stages. Unexpectedly however, we demonstrate here accumulation of OsHKT1;4 transcripts in the stem including internode II and peduncle in the reproductive growth stage. Moreover, phenotypic analysis of OsHKT1;4 RNAi plants in the vegetative growth stage revealed no profound influence on the growth and ion accumulation in comparison with WT plants upon salinity stress. However, imposition of salinity stress on the RNAi plants in the reproductive growth stage caused significant Na+ overaccumulation in aerial organs, in particular, leaf blades and sheaths. In addition, Na-22(+) tracer experiments using peduncles of RNAi and WT plants suggested xylem Na+ unloading by OsHKT1;4.
    Conclusions: Taken together, our results indicate a newly recognized function of OsHKT1;4 in Na+ exclusion in stems together with leaf sheaths, thus excluding Na+ from leaf blades of a japonica rice cultivar in the reproductive growth stage, but the contribution is low when the plants are in the vegetative growth stage.

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  • OsHKT2;2/1-mediated Na+ influx over K+ uptake in roots potentially increases toxic Na+ accumulation in a salt-tolerant landrace of rice Nona Bokra upon salinity stress

    Kei Suzuki, Alex Costa, Hideki Nakayama, Maki Katsuhara, Atsuhiko Shinmyo, Tomoaki Horie

    JOURNAL OF PLANT RESEARCH   129 ( 1 )   67 - 77   2016.1

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    HKT transporters are Na+-permeable membrane proteins, which mediate Na+ and K+ homeostasis in K+-depleted and saline environments in plants. Class II HKT transporters, a distinct subgroup found predominantly in monocots, are known to mediate Na+-K+ co-transport in principle. Here we report features of ion transport functions of No-OsHKT2;2/1, a class II transporter identified in a salt tolerant landrace of indica rice, Nona Bokra. We profiled No-OsHKT2;2/1 expression in organs of Nona Bokra plants with or without salinity stress. Dominant accumulation of the No-OsHKT2;2/1 transcript in K+-starved roots of Nona Bokra plants largely disappeared in response to 50 mM NaCl. We found that No-OsHKT2;2/1 expressed in the high-affinity K+ uptake deficient mutant of Saccharomyces cerevisiae and Xenopus laevis oocytes shows robust K+ selectivity even in the presence of a large amount of NaCl as reported previously. However, No-OsHKT2;2/1-expressing yeast cells exhibited Na+ hypersensitive growth under various concentrations of K+ and Na+ as the cells expressing Po-OsHKT2;2, a similar class II transporter from another salt tolerant indica rice Pokkali, when compared with the growth of cells harboring empty vector or cells expressing OsHKT2;4. The OsHKT2;4 protein expressed in Xenopus oocytes showed strong K+ selectivity in the presence of 50 mM NaCl in comparison with No-OsHKT2;2/1 and Po-OsHKT2;2. Together with apparent plasma membrane-localization of No-OsHKT2;2/1, these results point to possibilities that No-OsHKT2;2/1 could mediate destructive Na+ influx over K+ uptake in Nona Bokra plants upon salinity stress, and that a predominant physiological function of No-OsHKT2;2/1 might be the acquisition of Na+ and K+ in K+-limited environments.

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  • Yeast functional screen to identify genes conferring salt stress tolerance in Salicornia europaea

    Yoshiki Nakahara, Shogo Sawabe, Kenta Kainuma, Maki Katsuhara, Mineo Shibasaka, Masanori Suzuki, Kosuke Yamamoto, Suguru Oguri, Hikaru Sakamoto

    FRONTIERS IN PLANT SCIENCE   6   920   2015.10

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    Salinity is a critical environmental factor that adversely affects crop productivity. Halophytes have evolved various mechanisms to adapt to saline environments. Salicornia europaea L. is one of the most salt tolerant plant species. It does not have special salt-secreting structures like a salt gland or salt bladder, and is therefore a good model for studying the common mechanisms underlying plant salt tolerance. To identify candidate genes encoding key proteins in the mediation of salt tolerance in S. europaea, we performed a functional screen of a cDNA library in yeast. The library was screened for genes that allowed the yeast to grow in the presence of 1.3 M NaCl. We obtained three full-length S. europaea genes that confer salt tolerance. The genes are predicted to encode (1) a novel protein highly homologous to thaumatin-like proteins, (2) a novel coiled-coil protein of unknown function, and (3) a novel short peptide of 32 residues. Exogenous application of a synthetic peptide corresponding to the 32 residues improved salt tolerance of Arabidopsis. The approach described in this report provides a rapid assay system for large-scale screening of S. europaea genes involved in salt stress tolerance and supports the identification of genes responsible for such mechanisms. These genes may be useful candidates for improving crop salt tolerance by genetic transformation.

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  • Control of the Water Transport Activity of Barley HvTIP3;1 Specifically Expressed in Seeds

    Shigeko Utsugi, Mineo Shibasaka, Masahiko Maekawa, Maki Katsuhara

    PLANT AND CELL PHYSIOLOGY   56 ( 9 )   1831 - 1840   2015.9

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    Tonoplast intrinsic proteins (TIPs) are involved in the transport and storage of water, and control intracellular osmotic pressure by transporting material related to the water potential of cells. In the present study, we focused on HvTIP3;1 during the periods of seed development and desiccation in barley. HvTIP3;1 was specifically expressed in seeds. An immunochemical analysis showed that HvTIP3;1 strongly accumulated in the aleurone layers and outer layers of barley seeds. The water transport activities of HvTIP3;1 and HvTIP1;2, which also accumulated in seeds, were measured in the heterologous expression system of Xenopus oocytes. When they were expressed individually, HvTIP1;2 transported water, whereas HvTIP3;1 did not. However, HvTIP3;1 exhibited water transport activity when co-expressed with HvTIP1;2 in oocytes, and this activity was higher than when HvTIP1;2 was expressed alone. This is the first report to demonstrate that the water permeability of a TIP aquaporin was activated when co-expressed with another TIP. The split-yellow fluorescent protein (YFP) system in onion cells revealed that HvTIP3;1 interacted with HvTIP1;2 to form a heterotetramer in plants. These results suggest that HvTIP3;1 functions as an active water channel to regulate water movement through tissues during the periods of seed development and desiccation.

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  • Dynamic Regulation of the Root Hydraulic Conductivity of Barley Plants in Response to Salinity/Osmotic Stress

    Toshiyuki Kaneko, Tomoaki Horie, Yoshiki Nakahara, Nobuya Tsuji, Mineo Shibasaka, Maki Katsuhara

    PLANT AND CELL PHYSIOLOGY   56 ( 5 )   875 - 882   2015.5

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    Salinity stress significantly reduces the root hydraulic conductivity (Lp(r)) of several plant species including barley (Hordeum vulgare). Here we characterized changes in the Lp(r) of barley plants in response to salinity/osmotic stress in detail using a pressure chamber. Salt-tolerant and intermediate barley cultivars, K305 and Haruna-nijyo, but not a salt-sensitive cultivar, I743, exhibited characteristic time-dependent Lp(r) changes induced by 100 mM NaCl. An identical response was evoked by isotonic sorbitol, indicating that this phenomenon was triggered by osmotic imbalances. Further examination of this mechanism using barley cv. Haruna-nijyo plants in combination with the use of various inhibitors suggested that various cellular processes such as protein phosphorylation/dephosphorylation and membrane internalization appear to be involved. Interestingly, the three above-mentioned barley cultivars did not exhibit a remarkable difference in root cell sap osmolality under hypertonic conditions, in contrast to the case of Lp(r). The possible biological significance of the regulation of Lp(r) in barley plants upon salinity/osmotic stress is discussed.

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  • Osmotic stress decreases PIP aquaporin transcripts in barley roots but H2O2 is not involved in this process

    Maki Katsuhara, Nobuya Tsuji, Mineo Shibasaka, Sanjib Kumar Panda

    JOURNAL OF PLANT RESEARCH   127 ( 6 )   787 - 792   2014.11

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    Previous reports indicate that salt stress reduces the root hydraulic conductance and the expression of plasmamembrane-type aquaporins (PIPs). As a molecular mechanism for this phenomenon, the present study found evidence that the osmotic component, but probably not an ion-specific component, decreases PIP transcripts. Eight of ten PIP transcripts were reduced to less than half by 360 mM mannitol treatment for 12 h in comparison with control samples. A large decrease of HvPIP2; 1 protein was also recorded. This reduction of both transcripts and proteins of HvPIP2s should be physiologically effective for preventing or reducing dehydration at an initial phase of severe salt/osmotic stress. Root cell sap osmolality increased from 278 to 372 mOsm 24 h after 360 mM mannitol treatment. These steps can secure survival and growth recovery with water reabsorption in barley. Our data also suggest that H2O2 seems not to be the main cause of osmotic stress-induced transcriptional down-regulation within the concentrations (20-500 mu M) and time periods (24 h) examined, although H2O2 was previously proposed to be involved in the mechanisms of salinity/osmotic tolerance.

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  • Expression of root HvPIPs of barley seedling under chilling stress

    Rui-Jun Duan, Hui-Yan Xiong, Katsuhara Maki

    Zhiwu Shengli Xuebao/Plant Physiology Journal   50 ( 8 )   1203 - 1208   2014.8

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    The plant plasma membrane aquaporin (plasma membrane intrinsic proteins, PIPs) is a kind of membrane intrinsic proteins which has high selectivity and water transport. They play important roles in many physiological activities in plant growth and development. In this study, seedlings of barley variety 'Haruna-nijo' were treated with chilling stress. Expression of water channel protein PIPs of root in two processes which were stress period (4℃, 48 h) and recovery period (16℃, 48 h) were analyzed with quantitative real-time reverse transcriptase PCR (qRT-PCR). And root hydraulic conductivity (Lpr), root length and seedling height were analyzed at the same time. The results showed that: after 4℃ and 48 h stress, the growth of the barley seedlings was significantly inhibited, but root growth was no significant changed
    after 16℃, 48 h recovery, shoot height of stressed-seedling caught up that of control seedlings and root growth did not change significantly
    root hydraulic conductivity decreased in the period of chilling stress and increased rapidly in recovery period but there were no significant difference. The results of qRT-PCR showed: the highest expression were HvPIP1
    2 and HvPIP1
    3, the lowest expression were HvPIP1
    1 and HvPIP2
    3. Compared with the control group, HvPIPs expression were decreased in all after cold treatment, in which HvPIP1
    2, HvPIP1
    3, HvPIP1
    4, HvPIP1
    5, HvPIP2
    1, HvPIP2
    2 were down-regulated significantly. In recovery period, most of HvPIPs expression were increased, espeacally HvPIP1
    1, HvPIP1
    2, HvPIP1
    5, HvPIP2
    3 were up-regulated significantly, but HvPIP1
    4, HvPIP2
    5 were down-regulated. This study found that HvPIPs expressions of barley root were down-regulated overall in chilling stress and in recovery growth most HvPIPs were up-regulated. Combined with changes of root hydraulic conductivity and plant growth, the role of barley HvPIPs in chilling resistance reaction speculated that HvPIPs effected water absorption were different in different stages.

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  • CO2 Transport by PIP2 Aquaporins of Barley

    Izumi C. Mori, Jiye Rhee, Mineo Shibasaka, Shizuka Sasano, Toshiyuki Kaneko, Tomoaki Horie, Maki Katsuhara

    PLANT AND CELL PHYSIOLOGY   55 ( 2 )   251 - 257   2014.2

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    CO2 permeability of plasma membrane intrinsic protein 2 (PIP2) aquaporins of Hordeum vulgare L. was investigated. Five PIP2 members were heterologously expressed in Xenopus laevis oocytes. CO2 permeability was determined by decrease of cytosolic pH in CO2-enriched buffer using a hydrogen ion-selective microelectrode. HvPIP2; 1, HvPIP2; 2, HvPIP2; 3 and HvPIP2; 5 facilitated CO2 transport across the oocyte cell membrane. However, HvPIP2; 4 that is highly homologous to HvPIP2; 3 did not. The isoleucine residue at position 254 of HvPIP2; 3 was conserved in PIP2 aquaporins of barley, except HvPIP2; 4, which possesses methionine instead. CO2 permeability was lost by the substitution of the Ile254 of HvPIP2; 3 by methionine, while water permeability was not affected. These results suggest that PIP2 aquaporins are permeable to CO2. and the conserved isoleucine at the end of the E-loop is crucial for CO2 selectivity.

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  • Functional and molecular characteristics of rice and barley NIP aquaporins transporting water, hydrogen peroxide and arsenite

    Maki Katsuhara, Shizuka Sasano, Tomoaki Horie, Tadashi Matsumoto, Jiye Rhee, Mineo Shibasaka

    PLANT BIOTECHNOLOGY   31 ( 3 )   213 - U173   2014

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    Mercury-sensitive water transport activities were detected in seven NIP (Nodulin 26-like intrinsic protein) type aquaporins among eleven NIPs examined. Amino acid substitutions in rice OsNIP3;3 revealed that mercury-sensitivity depended on a histidine (but not on a cysteine) in apoplastic loop C in plant NIP aquaporins, although the cysteine is involved in the mercury-sensitivity of animal aquaporins. Rice OsNIP3;3 was also first identified as a unique aquaporin facilitating all water, hydrogen peroxide and arsenite transports. In rice OsNIP3;2, hydrogen peroxide and arsenite transport activities were detected, but water transport was not. Barley HvNIP1;2- or rice OsNIP2;1-expressing yeast cells showed the arsenite transport activity but not the H2O2 transport activity. The present work revealed novel molecular mechanisms of water and other low molecular weight compounds transport/selection in barley and rice NIP aquaporins, including the histidine-related mercury-sensitivity in the water transport of aquaporins.

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  • The photosynthetic response of tobacco plants overexpressing ice plant aquaporin McMIPB to a soil water deficit and high vapor pressure deficit

    Miki Kawase, Yuko T. Hanba, Maki Katsuhara

    JOURNAL OF PLANT RESEARCH   126 ( 4 )   517 - 527   2013.7

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    We investigated the photosynthetic capacity and plant growth of tobacco plants overexpressing ice plant (Mesembryanthemum crystallinum L.) aquaporin McMIPB under (1) a well-watered growth condition, (2) a well-watered and temporal higher vapor pressure deficit (VPD) condition, and (3) a soil water deficit growth condition to investigate the effect of McMIPB on photosynthetic responses under moderate soil and atmospheric humidity and water deficit conditions. Transgenic plants showed a significantly higher photosynthesis rate (by 48 %), higher mesophyll conductance (by 52 %), and enhanced growth under the well-watered growth condition than those of control plants. Decreases in the photosynthesis rate and stomatal conductance from ambient to higher VPD were slightly higher in transgenic plants than those in control plants. When plants were grown under the soil water deficit condition, decreases in the photosynthesis rate and stomatal conductance were less significant in transgenic plants than those in control plants. McMIPB is likely to work as a CO2 transporter, as well as control the regulation of stomata to water deficits.

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  • Overexpression of Alternative Oxidase Gene Confers Aluminum Tolerance by Altering the Respiratory Capacity and the Response to Oxidative Stress in Tobacco Cells

    Sanjib Kumar Panda, Lingaraj Sahoo, Maki Katsuhara, Hideaki Matsumoto

    MOLECULAR BIOTECHNOLOGY   54 ( 2 )   551 - 563   2013.6

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    Aluminum (Al) stress represses mitochondrial respiration and produces reactive oxygen species (ROS) in plants. Mitochondrial alternative oxidase (AOX) uncouples respiration from mitochondrial ATP production and may improve plant performance under Al stress by preventing excess accumulation of ROS. We tested respiratory changes and ROS production in isolated mitochondria and whole cell of tobacco (SL, ALT 301) under Al stress. Higher capacities of AOX pathways relative to cytochrome pathways were observed in both isolated mitochondria and whole cells of ALT301 under Al stress. AOX1 when studied showed higher AOX1 expression in ALT 301 than SL cells under stress. In order to study the function of tobacco AOX gene under Al stress, we produced transformed tobacco cell lines by introducing NtAOX1 expressed under the control of the cauliflower mosaic virus (CaMV) 35 S promoter in sensitive (SL) Nicotiana tabacum L. cell lines. The enhancement of endogenous AOX1 expression and AOX protein with or without Al stress was in the order of transformed tobacco cell lines &gt; ALT301 &gt; wild type (SL). A decreased respiratory inhibition and reduced ROS production with a better growth capability were the significant features that characterized AOX1 transformed cell lines under Al stress. These results demonstrated that AOX plays a critical role in Al stress tolerance with an enhanced respiratory capacity, reducing mitochondrial oxidative stress burden and improving the growth capability in tobacco cells.

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  • Water and CO2 permeability of SsAqpZ, the cyanobacterium Synechococcus sp. PCC7942 aquaporin

    Xiaodong Ding, Tadashi Matsumoto, Patrizia Gena, Chengwei Liu, Marialuisa Pellegrini-Calace, Shihua Zhong, Xiaoli Sun, Yanming Zhu, Maki Katsuhara, Ikuko Iwasaki, Yoshichika Kitagawa, Giuseppe Calamita

    Biology of the Cell   105 ( 3 )   118 - 128   2013.3

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    Background information: Cyanobacteria possess Aquaporin-Z (AqpZ) membrane channels which have been suggested to mediate the water efflux underlying osmostress-inducible gene expression and to be essential for glucose metabolism under photomixotrophic growth. However, preliminary observations suggest that the biophy-sical properties of transport and physiological meaning of AqpZ in such photosynthetic microorganisms are not yet completely assessed. Results: In this study, we used Xenopus laevis oocyte and proteoliposome systems to directly demonstrate the water permeability of the cyanobacterium Synechococcus sp. PCC7942 aquaporin, SsAqpZ. By an in vitro assay of intracellular acidification in yeast cells, SsAqpZ was found to transport also CO2. Consistent with this result, during the entire exponential phase of growth, Synechococcus SsAqpZ-null-mutant cells grew slower than the corresponding wild-type cells. This phenotype was stronger with higher levels of extracellular CO2. In line with the conversion of CO2 gas into HCO3- ions under alkaline conditions, the impairment in growth of the SsAqpZ-null strain was weaker in more alkaline culture medium. Conclusions: Cyanobacterial SsAqpZ may exert a pleiotropic function in addition to the already reported roles in macronutrient homeostasis and osmotic-stress response as it appears to constitute an important pathway in CO2 uptake, a fundamental step in photosynthesis. © 2013 Soçiété Française des Microscopies and Soçiété de Biologie Cellulaire de France.

    DOI: 10.1111/boc.201200057

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  • Aquaporin OsPIP1;1 promotes rice salt resistance and seed germination

    Chengwei Liu, Tatsuya Fukumoto, Tadashi Matsumoto, Patrizia Gena, Daniele Frascaria, Tomoyuki Kaneko, Maki Katsuhara, Shihua Zhong, Xiaoli Sun, Yanming Zhu, Ikuko Iwasaki, Xiaodong Ding, Giuseppe Calamita, Yoshichika Kitagawa

    PLANT PHYSIOLOGY AND BIOCHEMISTRY   63   151 - 158   2013.2

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    OsPIP1;1 is one of the most abundant aquaporins in rice leaves and roots and is highly responsible to environmental stresses. However, its biochemical and physiological functions are still largely unknown. The oocyte assay data showed OsPIP1;1 had lower water channel activity in contrast to OsPIP2;1. EGFP and immunoelectron microscopy studies revealed OsPIP1;1 was predominantly localized in not only plasma membrane but also in some ER-like intracellular compartments in the cells. OsPIP1;1 exhibited low water channel activity in Xenopus oocytes but coexpression of OsPIP2;1 significantly enhanced its water permeability. Stop-flow assay indicated that 10His-OsPIP1;1-reconstituted proteoliposomes had significantly higher water permeability than the control liposomes. Overexpression of OsPIP1;1 greatly altered many physiological features of transgenic plants in a dosage-dependent manner. Moderate expression of OsPIP1;1 increased rice seed yield, salt resistance, root hydraulic conductivity, and seed germination rate. This work suggests OsPIP1;1 functions as an active water channel and plays important physiological roles. Published by Elsevier Masson SAS.

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  • Salinity tolerance mechanisms in glycophytes: An overview with the central focus on rice plants. Reviewed

    Horie T, Karahara I, Katsuhara M

    Rice (New York, N.Y.)   5 ( 1 )   11 - 11   2012.12

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  • Influence of Low Air Humidity and Low Root Temperature on Water Uptake, Growth and Aquaporin Expression in Rice Plants

    Tsuneo Kuwagata, Junko Ishikawa-Sakurai, Hidehiro Hayashi, Kiyoshi Nagasuga, Keiko Fukushi, Arifa Ahamed, Katsuko Takasugi, Maki Katsuhara, Mari Murai-Hatano

    PLANT AND CELL PHYSIOLOGY   53 ( 8 )   1418 - 1431   2012.8

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    The effects of low air humidity and low root temperature (LRT) on water uptake, growth and aquaporin gene expression were investigated in rice plants. The daily transpiration of the plants grown at low humidity was 1.5- to 2-fold higher than that at high humidity. LRT at 13 degrees C reduced transpiration, and the extent was larger at lower humidity. LRT also reduced total dry matter production and leaf area expansion, and the extent was again larger at lower humidity. These observations suggest that the suppression of plant growth by LRT is associated with water stress due to decreased water uptake ability of the root. On the other hand, the net assimilation rate was not affected by low humidity and LRT, and water use efficiency was larger for LRT. We found that low humidity induced coordinated up-regulation of many PIP and TIP aquaporin genes in both the leaves and the roots. Expression levels of two root-specific aquaporin genes, OsPIP2;4 and OsPIP2;5, were increased significantly after 6 and 13 d of LRT exposure. Taken together, we discuss the possibility that aquaporins are part of an integrated response of this crop to low air humidity and LRT.

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  • Hydrogen peroxide permeability of plasma membrane aquaporins of Arabidopsis thaliana

    Cortwa Hooijmaijers, Ji Ye Rhee, Kyung Jin Kwak, Gap Chae Chung, Tomoaki Horie, Maki Katsuhara, Hunseung Kang

    JOURNAL OF PLANT RESEARCH   125 ( 1 )   147 - 153   2012.1

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    Although aquaporins have been known to transport hydrogen peroxide (H(2)O(2)) across cell membranes, the H(2)O(2)-regulated expression patterns and the permeability of every family member of the plasma membrane intrinsic protein (PIP) toward H(2)O(2) have not been determined. This study investigates the H(2)O(2)-regulated expression levels of all plasma membrane aquaporins of Arabidopsis thaliana (AtPIPs), and determines the permeability of every AtPIP for H(2)O(2) in yeast. Hydrogen peroxide treatment of Arabidopsis down-regulated the expression of AtPIP2 subfamily in roots but not in leaves, whereas the expression of AtPIP1 subfamily was not affected by H(2)O(2) treatment. The growth and survival of yeast cells that expressed AtPIP2;2, AtPIP2;4, AtPIP2;5, or AtPIP2;7 was reduced in the presence of H(2)O(2), while the growth of yeast cells expressing any other AtPIP family member was not affected by H(2)O(2). These results show that only certain isoforms of AtPIPs whose expression is regulated by H(2)O(2) treatment are permeable for H(2)O(2) in yeast cells, and suggest that the integrated regulation of aquaporin expression by H(2)O(2) and the capacity of individual aquaporin to transport H(2)O(2) are important for plant response to H(2)O(2).

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  • Functional characterization of a novel plasma membrane intrinsic protein2 in barley

    Mineo Shibasaka, Sizuka Sasano, Sigeko Utsugi, Maki Katsuhara

    Plant Signaling and Behavior   7 ( 12 )   1642 - 1646   2012

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    Water homeostasis is crucial to the growth and survival of plants. Plasma membrane intrinsic proteins (PIPs) have been shown to be primary channels mediating water uptake in plant cells. We characterized a novel PIP2 gene, HvPIP2
    8 in barley (Hordeum vulgare). HvPIP2
    8 shared 72-76% identity with other HvPIP2s and 74% identity with rice OsPIP2
    8. The gene was expressed in all organs including the shoots, roots and pistil at a similar level. When HvPIP2
    8 was transiently expressed in onion epidermal cells, it was localized to the plasma membrane. HvPIP2
    8 showed transport activity for water in Xenopus oocytes, however its interaction with HvPIP1
    2 was not observed. These results suggest that HvPIP2
    8 plays a role in water homeostasis although further functional analysis is required in future. © 2012 Landes Bioscience.

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  • Effect of nutrient deficiencies on the water transport properties in figleaf gourd plants

    Ji Ye Rhee, Gap Chae Chung, Maki Katsuhara, Sung-Ju Ahn

    HORTICULTURE ENVIRONMENT AND BIOTECHNOLOGY   52 ( 6 )   629 - 634   2011.12

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    Effects of nitrogen, phosphorus, and potassium deficiencies on water transport properties in figleaf gourd plants were studied. Plants were treated for different period of deficiency and physiological parameters such as stomatal conductance, photosynthesis and transpiration were measured. Cell and root pressure probes were utilized to measure turgor and root pressures, half-times of water exchange and hydraulic conductivities to analyze water transport properties. When plants were grown in nitrogen or phosphorus deficient nutrient solutions, they became insensitive to mercury, suggesting that aquaporin was closed resulting in reduced hydraulic conductivity. Inclusion of tungstate, however, restored the sensitivity of cells to mercury, indicating the importance of internal nutrient concentration, not the incoming nutrient supply. The hydrostatic hydraulic conductivity of roots grown in nitrogen deficient solution, representing apoplastic pathway of water transport, was reduced but this reduction was dramatically recovered by the application of tungstate, indicating the importance of nutrient availability from storage pools in relation to water status of plants.

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  • K+ Transport by the OsHKT2;4 Transporter from Rice with Atypical Na+ Transport Properties and Competition in Permeation of K+ over Mg2+ and Ca2+ Ions

    Tomoaki Horie, Dennis E. Brodsky, Alex Costa, Toshiyuki Kaneko, Fiorella Lo Schiavo, Maki Katsuhara, Julian I. Schroeder

    PLANT PHYSIOLOGY   156 ( 3 )   1493 - 1507   2011.7

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    Members of class II of the HKT transporters, which have thus far only been isolated from grasses, were found to mediate Na+-K+ cotransport and at high Na+ concentrations preferred Na+-selective transport, depending on the ionic conditions. But the physiological functions of this K+-transporting class II of HKT transporters remain unknown in plants, with the exception of the unique class II Na+ transporter OsHKT2;1. The genetically tractable rice (Oryza sativa; background Nipponbare) possesses two predicted K+-transporting class II HKT transporter genes, OsHKT2;3 and OsHKT2;4. In this study, we have characterized the ion selectivity of the class II rice HKT transporter OsHKT2;4 in yeast and Xenopus laevis oocytes. OsHKT2;4 rescued the growth defect of a K+ uptake-deficient yeast mutant. Green fluorescent protein-OsHKT2;4 is targeted to the plasma membrane in transgenic plant cells. OsHKT2;4-expressing oocytes exhibited strong K+ permeability. Interestingly, however, K+ influx in OsHKT2;4-expressing oocytes did not require stimulation by extracellular Na+, in contrast to other class II HKT transporters. Furthermore, OsHKT2;4-mediated currents exhibited permeabilities to both Mg2+ and Ca2+ in the absence of competing K+ ions. Comparative analyses of Ca2+ and Mg2+ permeabilities in several HKT transporters, including Arabidopsis thaliana HKT1;1 (AtHKT1;1), Triticum aestivum HKT2;1 (TaHKT2;1), OsHKT2;1, OsHKT2;2, and OsHKT2;4, revealed that only OsHKT2;4 and to a lesser degree TaHKT2;1 mediate Mg2+ transport. Interestingly, cation competition analyses demonstrate that the selectivity of both of these class II HKT transporters for K+ is dominant over divalent cations, suggesting that Mg2+ and Ca2+ transport via OsHKT2;4 may be small and would depend on competing K+ concentrations in plants.

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  • Mechanisms of Water Transport Mediated by PIP Aquaporins and Their Regulation Via Phosphorylation Events Under Salinity Stress in Barley Roots

    Tomoaki Horie, Toshiyuki Kaneko, Genki Sugimoto, Shizuka Sasano, Sanjib Kumar Panda, Mineo Shibasaka, Maki Katsuhara

    PLANT AND CELL PHYSIOLOGY   52 ( 4 )   663 - 675   2011.4

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    Water homeostasis is crucial to the growth and survival of plants under water-related stress. Plasma membrane intrinsic proteins (PIPs) have been shown to be primary channels mediating water uptake in plant cells. Here we report the water transport activity and mechanisms for the regulation of barley (Hordeum vulgare) PIP aquaporins. HvPIP2 but not HvPIP1 channels were found to show robust water transport activity when expressed alone in Xenopus laevis oocytes. However, the co-expression of HvPIP1 with HvPIP2 in oocytes resulted in significant increases in activity compared with the expression of HvPIP2 alone, suggesting the participation of HvPIP1 in water transport together with HvPIP2 presumably through heteromerization. Severe salinity stress (200 mM NaCl) significantly reduced root hydraulic conductivity (Lp(r)) and the accumulation of six of 10 HvPIP mRNAs. However, under relatively mild stress (100 mM NaCl), only a moderate reduction in Lp(r) with no significant difference in HvPIP mRNA levels was observed. Sorbitol-mediated osmotic stress equivalent to 100 and 200 mM NaCl induced nearly identical Lp(r) reductions in barley roots. Furthermore, the water transport activity in intact barley roots was suggested to require phosphorylation that is sensitive to a kinase inhibitor, staurosporine. HvPIP2s also showed water efflux activity in Xenopus oocytes, suggesting a potential ability to mediate water loss from cells under hypertonic conditions. Water transport via HvPIP aquaporins and the significance of reductions of Lp(r) in barley plants during salinity stress are discussed.

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  • Rice sodium-insensitive potassium transporter, OsHAK5, confers increased salt tolerance in tobacco BY2 cells

    Tomoaki Horie, Mitsuo Sugawara, Tomoyuki Okada, Koichiro Taira, Pulla Kaothien-Nakayama, Maki Katsuhara, Atsuhiko Shinmyo, Hideki Nakayama

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   111 ( 3 )   346 - 356   2011.3

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    Potassium ion (K(+)) plays vital roles in many aspects of cellular homeostasis including competing with sodium ion (Na(+)) during potassium starvation and salt stress. Therefore, one way to engineer plant cells with improved salt tolerance is to enhance K(+) uptake activity of the cells, while keeping Na(+) out during salt stress. Here, in search for Na(+)-insensitive transporter for this purpose, bacterial expression system was used to characterize two K(+) transporters, OsHAK2 and OsHAK5, isolated from rice (Oryza sativa cv. Nipponbare). The two OsHAK transporters are members of a KT/HAK/KUP transporter family, which is one of the major K(+) transporter families in bacteria, fungi and plants. When expressed in an Escherichia cola K(+) transport mutant strain LB2003, both OsHAK transporters rescued the growth defect in K(+)-limiting conditions by significantly increasing the K(+) content of the cells. Under the condition with a large amount of extracellular Na(+), we found that OsHAK5 functions as a Na(+)-insensitive K(+) transporter, while OsHAK2 is sensitive to extracellular Na(+) and exhibits higher Na(+) over K(+) transport activities. Moreover, constitutive expression of OsHAK5 in cultured-tobacco BY2 (Nicotiana tabacum cv. Bright Yellow 2) cells enhanced the accumulation of K(+) but not Na(+) in the cells during salt stress and conferred increased salt tolerance to the cells. Transient expression experiment indicated that OsHAK5 is localized to the plant plasma membrane. These results suggest that the plasma-membrane localized Na(+) insensitive K(+) transporters, similar to OsHAK5 identified here, could be used as a tool to enhance salt tolerance in plant cells. (C) 2010, The Society for Biotechnology, japan. All rights reserved.

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  • Hormonal treatment of the bark of rubber trees (Hevea brasiliensis) increases latex yield through latex dilution in relation with the differential expression of two aquaporin genes

    Kessarin Tungngoen, Unchera Viboonjun, Panida Kongsawadworakul, Maki Katsuhara, Jean-Louis Julien, Soulaiman Sakr, Herve Chrestin, Jarunya Narangajavana

    JOURNAL OF PLANT PHYSIOLOGY   168 ( 3 )   253 - 262   2011.2

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    Natural rubber is synthesized in laticifers in the inner liber of the rubber tree (Hevea brasiliensis). Upon bark tapping, the latex is expelled due to fiber turgor pressure. The mature laticifers are devoid of plasmodesmata; therefore a corresponding decrease-in the total latex solid content is likely to occur due to water influx inside the laticifers. Auxins and ethylene used as efficient yield stimulants in mature untapped rubber trees, but, bark treatments with abscisic acid (ABA) and salicylic acid (SA) could also induce a transient increase latex yield. We recently reported that there are three aquaporin genes, HbPIP2;1, HbT1P1;1 and HbPIP1;1, that are regulated differentially after ethylene bark treatment. HbPIP2:1 was up-regulated in both the laticifers and the inner liber tissues, whereas HbT1P1:1 was up-regulated in the latex cells, but very markedly down-regulated in the inner liber tissues. Conversely, HbPIP1:1 was down-regulated in both tissues. In the present study, HbPIP2;1 and HbT1P1:1 showed a similar expression in response to auxin, ABA and SA, as seen in ethylene stimulation, while HbPIP1;1 was slightly regulated by auxin, but neither by ABA nor SA. The analysis of the HbPIP1:1 promoter region indicated the presence of only ethylene and auxin responsive elements. In addition, the poor efficiency of this HbPIP1;1 in increasing plasmalemma water conductance was confirmed in Xenopus oocytes. Thus, an increase in latex yield in response to all of these hormones was proposed to be the major function of aquaporins, HbPIP2:1 and HbTIP1;1. This study emphasized that the circulation of water between the laticifers and their surrounding tissues that result in latex dilution, as well as the probable maintenance of the liber tissues turgor pressure, favor the prolongation of latex flow. (C) 2010 Elsevier GmbH. All rights reserved.

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  • Abiotic stresses modulate expression of major intrinsic proteins in barley (Hordeum vulgare)

    Ayalew Ligaba, Maki Katsuhara, Mineo Shibasaka, Gemechis Djira

    COMPTES RENDUS BIOLOGIES   334 ( 2 )   127 - 139   2011.2

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    In one of the most important crops, barley (Hordeum vulgare L), gene expression and physiological roles of most major intrinsic proteins (MIPs) remained to be elucidated. Here we studied expression of five tonoplast intrinsic protein isoforms (HvTIP1;2, HvTIP2:1, HvTIP2;2,HvTIP2;3 and HvTIP4:1), a NOD26-like intrinsic protein (HvNIP2;1) and a plasma membrane intrinsic protein (HvPIP2;1) by using the quantitative real-time RT-PCR. Five-day-old seedlings were exposed to abiotic stresses (salt, heavy metals and nutrient deficiency), abscisic acid (ABA) and gibberellic acid (GA) for 24 h. Treatment with 100 mM NaCl, 0.1 mM ABA and 1 mM GA differentially regulated gene expression in roots and shoots. Nitrogen and prolonged P-deficiency downregulated expression of most MIP genes in roots. Intriguingly, gene expression was restored to the values in the control three days after nutrient supply was resumed. Heavy metals (0.2 mM each of Cd, Cu, Zn and Cr) downregulated the transcript levels by 60-80% in roots, whereas 0.2 mM Hg upregulated expressions of most genes in roots. This was accompanied by a 45% decrease in the rate of transpiration. In order to study the physiological role of the MIPs, cDNA of three genes (HvTIP2:1, HvTIP2:3 and HvNIP2;1) have been cloned and heterologous expression was performed in Xenopus laevis oocytes. Osmotic water permeability was determined by a swelling assay. However, no water uptake activity was observed for the three proteins. Hence, the possible physiological role of the proteins is discussed. (C) 2010 Academie des sciences. Published by Elsevier Masson SAS. All rights reserved.

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  • Early response in water relations influenced by NaCl reflects tolerance or sensitivity of barley plants to salinity stress via aquaporins

    Maki Katsuhara, Ji Ye Rhee, Genki Sugimoto, Gap Chae Chung

    SOIL SCIENCE AND PLANT NUTRITION   57 ( 1 )   50 - 60   2011.2

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    Barley varieties, K305 and I743, which are sodium chloride (NaCl) tolerant and sensitive respectively, were hydroponically grown to determine the short-term effects of NaCl on the cell water relations in roots using a cell pressure probe, and on the transcript levels of 10 barley PIP aquaporin genes (HvPIPs) in roots. Stomatal conductance, as an indicator of sensitivity to NaCl, was decreased to less than half values of control upon exposure to 100 mmol L-1 NaCl for 24 h in I743 whereas tolerant variety, K305, was able to maintain original conductance. Osmotic half-times of water exchange in cortical cells allowed for a clear distinction between the two varieties up to 200 mmol L-1 NaCl. With treatment duration of up to 12 h with 100 mmol L-1 NaCl, the elastic modulus was reduced in I743 but increased in K305. Hydrostatic half-times of water exchange in K305 increased rapidly, whereas this value remained unchanged in I743. Application of abscisic acid (ABA) after 1 h NaCl treatment restored the hydraulic conductivity of cells (Lp) in K305 but not in I743 whereas the opposite results were obtained when mercury chloride (HgCl2) was applied, verifying the contrasting gating response of aquaporins in two varieties. Reduced expression of HvPIPs was consistent with the reduction of hydraulic conductivity of both varieties after 24 h NaCl, but without any significant differences between them, indicating the importance of the activities of existing aquaporins rather than de novo synthesis to cope with short-term effects of salt stress.

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  • Water and H2O2 Permeability of Aquaporins Isolated from Cucumber and Fig leaf Gourd

    Ji Ye Rhee, Maki Katsuhara, Hunseung Kang, Gap Chae Chung

    HORTICULTURE ENVIRONMENT AND BIOTECHNOLOGY   51 ( 3 )   167 - 172   2010.6

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    Cucumber aquaporin gene, CsPIP1;1 and figleaf gourd aquaporin gene CfPIP2;1 were injected to Xenopus oocytes in order to study water transport activity. While CsPIP1;1 did not transport any water and was not responsive to mercury, oocyte injected with CfPIP2;1 rapidly increased their volume without saturation indicating active water transport. Calculation of water permeability of oocytes clearly distinguished the two genes. In response to exogenous application of H2O2, only CfPIP2;1 responded, reducing osmotic water permeability. Transgenic Arabidopsis plants overexpressing aquaporin genes did not respond to applied H2O2.

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  • Insights into the salt tolerance mechanism in barley (Hordeum vulgare) from comparisons of cultivars that differ in salt sensitivity

    Ayalew Ligaba, Maki Katsuhara

    JOURNAL OF PLANT RESEARCH   123 ( 1 )   105 - 118   2010.1

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    Although barley (Hordeum vulgare L.) is a salt-tolerant crop, the underlying physiological and molecular mechanisms of salt tolerance remain to be elucidated. Therefore, we investigated the response of salt-tolerant (K305) and salt-sensitive (I743) cultivars to salt stress at both physiological and molecular levels. Salt treatment increased xylem sap osmolarity, which was attributed primarily to a rise in Na(+) and Cl(-) concentration; enhanced accumulation of the ions in shoots; and reduced plant growth more severely in I743 than K305. The concentration of K(+) in roots and shoots decreased during 8 h of salt treatment in both cultivars but with no marked difference between cultivars. Hence, the severe growth reduction in I743 is attributed to the elevated levels of (mainly) Na(+) in shoots. Analysis of gene expression using quantitative RT-PCR showed that transcripts of K(+)-transporters (HvHAK1 and HvAKT1), vacuolar H(+)-ATPase and inorganic pyrophosphatase (HvHVA/68 and HvHVP1) were more abundant in shoots of K305 than in shoots of I743. Expression of HvHAK1 and Na(+)/H(+) antiporters (HvNHX1, HvNHX3 and HvNHX4) was higher in roots of K305 than in I743 with prolonged exposure to salt. Taken together, these results suggest that the better performance of K305 compared to I743 during salt stress may be related to its greater ability to sequester Na(+) into sub-cellular compartments and/or maintain K(+) homeostasis.

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  • Differential Sodium and Potassium Transport Selectivities of the Rice OsHKT2;1 and OsHKT2;2 Transporters in Plant Cells

    Xuan Yao, Tomoaki Horie, Shaowu Xue, Ho-Yin Leung, Maki Katsuhara, Dennis E. Brodsky, Yan Wu, Julian I. Schroeder

    PLANT PHYSIOLOGY   152 ( 1 )   341 - 355   2010.1

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    Na+ and K+ homeostasis are crucial for plant growth and development. Two HKT transporter/channel classes have been characterized that mediate either Na+ transport or Na+ and K+ transport when expressed in Xenopus laevis oocytes and yeast. However, the Na+/K+ selectivities of the K+-permeable HKT transporters have not yet been studied in plant cells. One study expressing 5' untranslated region-modified HKT constructs in yeast has questioned the relevance of cation selectivities found in heterologous systems for selectivity predictions in plant cells. Therefore, here we analyze two highly homologous rice (Oryza sativa) HKT transporters in plant cells, OsHKT2;1 and OsHKT2;2, that show differential K+ permeabilities in heterologous systems. Upon stable expression in cultured tobacco (Nicotiana tabacum) Bright-Yellow 2 cells, OsHKT2;1 mediated Na+ uptake, but little Rb+ uptake, consistent with earlier studies and new findings presented here in oocytes. In contrast, OsHKT2; 2 mediated Na+-K+ cotransport in plant cells such that extracellular K+ stimulated OsHKT2;2-mediated Na+ influx and vice versa. Furthermore, at millimolar Na+ concentrations, OsHKT2; 2 mediated Na+ influx into plant cells without adding extracellular K+. This study shows that the Na+/K+ selectivities of these HKT transporters in plant cells coincide closely with the selectivities in oocytes and yeast. In addition, the presence of external K+ and Ca2+ down-regulated OsHKT2;1-mediated Na+ influx in two plant systems, Bright-Yellow 2 cells and intact rice roots, and also in Xenopus oocytes. Moreover, OsHKT transporter selectivities in plant cells are shown to depend on the imposed cationic conditions, supporting the model that HKT transporters are multi-ion pores.

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  • A bacterial biosensor for oxidative stress using the constitutively expressed redox-sensitive protein roGFP2. Reviewed International journal

    Arias-Barreiro CR, Okazaki K, Koutsaftis A, Inayat-Hussain SH, Tani A, Katsuhara M, Kimbara K, Mori IC

    Sensors (Basel, Switzerland)   10 ( 7 )   6290 - 6306   2010

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    A highly specific, high throughput-amenable bacterial biosensor for chemically induced cellular oxidation was developed using constitutively expressed redox-sensitive green fluorescent protein roGFP2 in E. coli (E. coli-roGFP2). Disulfide formation between two key cysteine residues of roGFP2 was assessed using a double-wavelength ratiometric approach. This study demonstrates that only a few minutes were required to detect oxidation using E. coli-roGFP2, in contrast to conventional bacterial oxidative stress sensors. Cellular oxidation induced by hydrogen peroxide, menadione, sodium selenite, zinc pyrithione, triphenyltin and naphthalene became detectable after 10 seconds and reached the maxima between 80 to 210 seconds, contrary to Cd(2+), Cu(2+), Pb(2+), Zn(2+) and sodium arsenite, which induced the oxidation maximum immediately. The lowest observable effect concentrations (in ppm) were determined as 1.0 × 10(-7) (arsenite), 1.0 × 10(-4) (naphthalene), 1.0 × 10(-4) (Cu(2+)), 3.8 × 10(-4) (H(2)O(2)), 1.0 × 10(-3) (Cd(2+)), 1.0 × 10(-3) (Zn(2+)), 1.0 × 10(-2) (menadione), 1.0 (triphenyltin), 1.56 (zinc pyrithione), 3.1 (selenite) and 6.3 (Pb(2+)), respectively. Heavy metal-induced oxidation showed unclear response patterns, whereas concentration-dependent sigmoid curves were observed for other compounds. In vivo GSH content and in vitro roGFP2 oxidation assays together with E. coli-roGFP2 results suggest that roGFP2 is sensitive to redox potential change and thiol modification induced by environmental stressors. Based on redox-sensitive technology, E. coli-roGFP2 provides a fast comprehensive detection system for toxicants that induce cellular oxidation.

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  • Regulation mechanism of water permeability under hypertonic stress in barley root

    Kaneko Toshiyuki, Horie Tomoaki, Shibasaka Mineo, Katsuhara Maki

    Plant and Cell Physiology Supplement   2010   0034 - 0034   2010

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    The salt stress is one of the representative abiotic stresses that decreases growth and productivity of crops plants. The root water permeability of three barley (Hordeum Vulgare) cultivars that show a difference in the salt sensitivity was measured under salinity stress using a pressure chamber. Water permeability of salt sensitive variety did not show any difference in response to salt stress, in contrast to significant reductions of water permeability found in more salt tolerant varieties. Root water permeability of tolerant cultivars exposed to 100 mM NaCl was found to be extremely down-regulated after 1h. Interestingly, however, water permeability was partially recovered after 4 h, which was followed by a re-down-regulation afterwards. Further measurements using various inhibitors indicated that at least protein phosphorylation and recycling plays an essential role in the regulation mechanism.

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  • Involvement of HbPIP2;1 and HbTIP1;1 Aquaporins in Ethylene Stimulation of Latex Yield through Regulation of Water Exchanges between Inner Liber and Latex Cells in Hevea brasiliensis

    Kessarin Tungngoen, Panida Kongsawadworakul, Unchera Viboonjun, Maki Katsuhara, Nicole Brunel, Soulaiman Sakr, Jarunya Narangajavana, Herve Chrestin

    PLANT PHYSIOLOGY   151 ( 2 )   843 - 856   2009.10

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    Natural rubber is synthesized in specialized articulated cells (laticifers) located in the inner liber of Hevea brasiliensis. Upon bark tapping, the laticifer cytoplasm (latex) is expelled due to liber tissue turgor pressure. In mature virgin (untapped) trees, short-term kinetic studies confirmed that ethylene, the rubber yield stimulant used worldwide, increased latex yield, with a concomitant decrease in latex total solid content, probably through water influx in the laticifers. As the mature laticifers are devoid of plasmodesmata, the rapid water exchanges with surrounding liber cells probably occur via the aquaporin pathway. Two full-length aquaporin cDNAs (HbPIP2;1 and HbTIP1;1, for plasma membrane intrinsic protein and tonoplast intrinsic protein, respectively) were cloned and characterized. The higher efficiency of HbPIP2;1 than HbTIP1;1 in increasing plasmalemma water conductance was verified in Xenopus laevis oocytes. HbPIP2;1 was insensitive to HgCl(2). In situ hybridization demonstrated that HbPIP2;1 was expressed in all liber tissues in the young stem, including the laticifers. HbPIP2;1 was up-regulated in both liber tissues and laticifers, whereas HbTIP1;1 was down-regulated in liber tissues but up-regulated in laticifers in response to bark Ethrel treatment. Ethylene-induced HbPIP2;1 up-regulation was confirmed by western-blot analysis. The promoter sequences of both genes were cloned and found to harbor, among many others, ethylene-responsive and other chemical-responsive (auxin, copper, and sulfur) elements known to increase latex yield. Increase in latex yield in response to ethylene was emphasized to be linked with water circulation between the laticifers and their surrounding tissues as well as with the probable maintenance of liber tissue turgor, which together favor prolongation of latex flow.

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  • Biomarkers of Green Roof Vegetation: Anthocyanin and Chlorophyll as Stress Marker Pigments for Plant Stresses of Rooftop Environments.

    Mori, I.C, Utsugi,S, Tanakamaru, S, Tani, A, Enomoto, T, Katsuhara, M

    Journal of Environmental Engineering and Management   19   21 - 27   2009

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  • New production method for greening concrete blocks.

    Katsuhara M

    Environment Solution Technology   8 ( 3 )   32 - 36   2009

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  • Function and regulation of PIP-type water channels in roots of barley under salinity stress

    Horie Tomoaki, Kaneko Toshiyuki, Sugimoto Genki, Shibasaka Mineo, Katsuhara Maki

    Plant and Cell Physiology Supplement   2009   S0029 - S0029   2009

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    The plasma membrane intrinsic protein (PIP) forms the pore for water transport as a water channel in plant cells. We have studied the molecular mechanism of water transport in roots of barley (Hordeum Vulgare cv. Haruna-nijo). Quantitative RT-PCR analyses using roots of barley plants revealed significant reductions in the accumulation of 6 out of 10 HvPIP mRNA in response to 200 mM NaCl. Characterization of water permeability of barely roots using a pressure chamber showed that water permeability was extremely down-regulated in 200 mM NaCl condition. Furthermore, phosphorylation was found to regulate water transport activity. HvPIP1 proteins did not show apparent water transport activity when they were expressed alone in Xenopus laevis oocytes. However, co-expression of HvPIP1 proteins with HvPIP2 proteins was shown to enhance water transport activity of HvPIP2 proteins. The regulation mechanism of water transport via HvPIP proteins in barley roots under salinity stress will be discussed.

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  • Characterization of four plasma membrane aquaporins in tulip petals: A putative homolog is regulated by phosphorylation

    Abul Kalam Azad, Maki Katsuhara, Yoshihiro Sawa, Takahiro Ishikawa, Hitoshi Shibata

    PLANT AND CELL PHYSIOLOGY   49 ( 8 )   1196 - 1208   2008.8

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    We suggested previously that temperature-dependent tulip (Tulipa gesneriana) petal movement that is concomitant with water transport is regulated by reversible phosphorylation of an unidentified plasma membrane intrinsic protein (PIP). In this study, four full-length cDNAs of PIPs from tulip petals were identified and cloned. Two PIPs, namely TgPIP1;1 and TgPIP1;2, are members of the PIP1 subfamily, and the remaining two PIPs, namely TgPIP2;1 and TgPIP2;2, belong to the PIP2 subfamily of aquaporins and were named according to the nomenclature of PIP genes in plants. Of these four homologs, only TgPIP2;2 displayed significant water channel activity in the heterologous expression assay using Xenopus laevis oocytes. The water channel activity of this functional isoform was abolished by mercury and was affected by inhibitors of protein kinase and protein phosphatase. Using a site-directed mutagenesis approach to substitute several serine residues with alanine, and assessing water channel activity using the methylotrophic yeast Pichia pastoris expression assay, we showed that Ser35, Ser116 and Ser274 are the putative phosphorylation sites of TgPIP2;2. Real-time reverse transcriptionPCR analysis revealed that the transcript levels of TgPIP1;1 and TgPIP1;2 in tulip petals, stems, leaves, bulbs and roots are very low when compared with those of TgPIP2;1 and TgPIP2;2. The transcript level of TgPIP2;1 is negligible in roots, and TgPIP2;2 is ubiquitously expressed in all organs with significant transcript levels. From the data reported herein, we suggest that TgPIP2;2 might be modulated by phosphorylation and dephosphorylation for regulating water channel activity, and may play a role in transcellular water transport in all tulip organs.

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  • Barley plasma membrane intrinsic proteins (PIP aquaporins) as water and CO(2) transporters

    Maki Katsuhara, Yuko T. Hanba

    PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY   456 ( 4 )   687 - 691   2008.7

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    We identified barley aquaporins and demonstrated that one, HvPIP2;1, transports water and CO(2). Regarding water homeostasis in plants, regulations of aquaporin expression were observed in many plants under several environmental stresses. Under salt stress, a number of plasma membrane-type aquaporins were down-regulated, which can prevent continuous dehydration resulting in cell death. The leaves of transgenic rice plants that expressed the largest amount of HvPIP2;1 showed a 40% increase in internal CO(2) conductance compared with leaves of wild-type rice plants. The rate of CO(2) assimilation also increased in the transgenic plants. The goal of our plant aquaporin research is to determine the key aquaporin species responsible for water and CO(2) transport, and to improve plant water relations, stress tolerance, CO(2) uptake or assimilation, and plant productivity via molecular breeding of aquaporins.

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  • Drought stress alters water relations and expression of PIP-type aquaporin genes in Nicotiana tabacum plants

    Majid Mahdieh, Akbar Mostajeran, Tomoaki Horie, Maki Katsuhara

    PLANT AND CELL PHYSIOLOGY   49 ( 5 )   801 - 813   2008.5

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    Plasma membrane intrinsic proteins (PIPs), a type of aquaporins, mediate water transport in many plant species. In this study, we investigated the relationship between the functions of PIP-type water channels and water relations of tobacco plants (Nicotiana tabacum cv. Samsun) under drought stress. Drought stress treatments have led to reductions in the stomatal conductance, transpiration, water potential and turgor pressure in leaves, and also the sap flow rate and osmotic hydraulic conductance in roots. In contrast, leaf osmotic pressure was increased in response to drought stress. Interestingly, the accumulation of NtPIP1;1 and NtPIP2;1 transcripts was significantly decreased, but only that of the NtAQP1 transcript was increased under drought stress. Functional analysis using Xenopus laevis oocytes revealed that NtPIP2;1 shows marked water transport activity, but the activities of NtAQP1 and NtPIP1;1 are weak or almost negligible, respectively, when expressed alone. However, co-expression of NtPIP1;1 with NtPIP2;1 significantly enhanced water transport activity compared with that of NtPIP1;1- or NtPIP2;1-expressing oocytes, suggesting that these two aquaporins may function as a water channel, forming a heterotetramer. Heteromerization of NtPIP1;1 and NtPIP2;1 was also suggested by co-expression analyses of NtPIP1;1-GFP (green fluorescent protein) and NtPIP2;1 in Xenopus oocytes. Re-watering treatments recovered water relation parameters and the accumulation of the three NtPIP transcripts to levels similar to control conditions. These results suggest that NtPIP1;1 and NtPIP2;1 play an important role in water transport in roots, and that expression of NtPIP1;1 and NtPIP2;1 is down-regulated in order to reduce osmotic hydraulic conductance in the roots of tobacco plants under drought stress.

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  • Presence of aquaporin and V-ATPase on the contractile vacuole of Amoeba proteus

    Eri Nishihara, Etsuo Yokota, Akira Tazaki, Hidefurni Orii, Maki Katsuhara, Kensuke Kataoka, Hisako Igarashi, Yoshinori Moriyama, Teruo Shimmen, Seiji Sonobe

    BIOLOGY OF THE CELL   100 ( 3 )   179 - 188   2008.3

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    Background information. The results of water permeability measurements suggest the presence of an AQP (aquaporin) in the membrane of the CV (contractile vacuole) in Amoeba proteus [Nishihara, Shimmen and Sonobe (2004) Cell Struct. Funct. 29, 85-90].
    Results. In the present study, we cloned an AQP gene from A. proteus [ApAQP (A. proteus AQP)] that encodes a 295-amino-acid protein. The protein has six putative TMs (transmembrane domains) and two NPA (Asn-Pro-Ala) motifs, which are conserved among various AQPs and are thought to be involved in the formation of water channels that span the lipid bilayer. Using Xenopus oocytes, we have demonstrated that the ApAQP protein product can function as a water channel. Immuncifluorescence microscopy with anti-ApAQP antibody revealed that ApAQP is detected on the CV membrane and on the vesicles around the CV. The presence of V-ATPase (vacuolar H+-ATPase) on the vesicle membrane around the CV was also detected.
    Conclusions. Our data on ApAQP allow us to provide the first informed explanation of the high water permeability of the CV membrane in amoeba. Moreover, the results suggest that vesicles possessing V-ATPase are involved in generating an osmotic gradient. Based on our findings, we propose a new hypothesis for the mechanism of CV function.

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  • Female mating receptivity inhibited by injection of male-derived extracts in Callosobruchus chinensis

    Takashi Yamane, Yoshinobu Kimura, Maki Katsuhara, Takahisa Miyatake

    JOURNAL OF INSECT PHYSIOLOGY   54 ( 2 )   501 - 507   2008.2

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    The effects of male-derived extracts on female receptivity to remating were investigated in Callosobruchus chinensis (Coleoptera: Bruchidae). Injection of aqueous extracts of male reproductive tracts into the abdomen of females reduced receptivity. When aqueous extracts of male reproductive tracts were divided to three molecular weight (MW) fractions by ultrafiltration: &lt; 3, 3-14, and &gt; 14 kDa, the filtrate containing MW substances &lt; 3 kDa reduced female receptivity 3 h and I day after injection, whereas the fraction containing MW substances &gt; 14 kDa inhibited receptivity 2 and 4 days after injection. Finally, male reproductive tract organs were divided into accessory gland, seminal vesicle, and testis. Aqueous extracts of testis reduced receptivity of females on the second day and at 3 h, and aqueous extracts of accessory gland reduced receptivity of females on the second day after injection. On the other hand, aqueous extracts of seminal vesicle did not reduce female receptivity. The results indicate that more than one mechanism may be involved in producing the effects of male-derived substances on female receptivity; low MW male-derived substances, which possibly exist in testis, cause short-term inhibition, while high MW substances, which possibly exist in the accessory gland, inhibit female mating later than low MW substances in C chinensis. (c) 2007 Elsevier Ltd. All rights reserved.

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  • Expanding roles of plant aquaporins in plasma membranes and cell organelles

    KATSUHARA M.

    Funct. Plant Biol.   35 ( 1 )   1 - 14   2008

  • Barley root hydraulic conductivity and aquaporins expression in relation to salt tolerance

    Maki Katsuhara, Mineo Shibasaka

    SOIL SCIENCE AND PLANT NUTRITION   53 ( 4 )   466 - 470   2007.8

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    Root hydraulic conductivity (Lp(r)) and aquaporins were investigated in barley seedlings under salt stress of 100 mmol L-1 NaCl in hydroponic culture for 24 and 48 h. No reduction in root elongation was observed in stressed seedlings, and more that 75% of root and shoot growth was maintained under salt stress of 100 mmol L-1 NaCl. A slight increase in HvPIP2;1 transcript, which was the most abundant of the three aquaporins investigated, was observed. No significant changes in LPr and HvPIP2;1 protein were detected. The possible reason for these complicated results and the function of aquaporins in LPr under salt stress are discussed.

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  • An aluminum-activated citrate transporter in barley.

    Jun Furukawa, Naoki Yamaji, Hua Wang, Namiki Mitani, Yoshiko Murata, Kazuhiro Sato, Maki Katsuhara, Kazuyoshi Takeda, Jian Feng Ma

    Plant & cell physiology   48 ( 8 )   1081 - 91   2007.8

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    Soluble ionic aluminum (Al) inhibits root growth and reduces crop production on acid soils. Al-resistant cultivars of barley (Hordeum vulgare L.) detoxify Al by secreting citrate from the roots, but the responsible gene has not been identified yet. Here, we identified a gene (HvAACT1) responsible for the Al-activated citrate secretion by fine mapping combined with microarray analysis, using an Al-resistant cultivar, Murasakimochi, and an Al-sensitive cultivar, Morex. This gene belongs to the multidrug and toxic compound extrusion (MATE) family and was constitutively expressed mainly in the roots of the Al-resistant barley cultivar. Heterologous expression of HvAACT1 in Xenopus oocytes showed efflux activity for (14)C-labeled citrate, but not for malate. Two-electrode voltage clamp analysis also showed transport activity of citrate in the HvAACT1-expressing oocytes in the presence of Al. Overexpression of this gene in tobacco enhanced citrate secretion and Al resistance compared with the wild-type plants. Transiently expressed green fluorescent protein-tagged HvAACT1 was localized at the plasma membrane of the onion epidermal cells, and immunostaining showed that HvAACT1 was localized in the epidermal cells of the barley root tips. A good correlation was found between the expression of HvAACT1 and citrate secretion in 10 barley cultivars differing in Al resistance. Taken together, our results demonstrate that HvAACT1 is an Al-activated citrate transporter responsible for Al resistance in barley.

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  • An efflux transporter of silicon in rice

    Jian Feng Ma, Naoki Yamaji, Namiki Mitani, Kazunori Tamai, Saeko Konishi, Toru Fujiwara, Maki Katsuhara, Masahiro Yano

    NATURE   448 ( 7150 )   209 - U12   2007.7

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    Silicon is an important nutrient for the optimal growth and sustainable production of rice(1-4). Rice accumulates up to 10% silicon in the shoot, and this high accumulation is required to protect the plant from multiple abiotic and biotic stresses(1-5). A gene, Lsi1, that encodes a silicon influx transporter has been identified in rice(6). Here we describe a previously uncharacterized gene, low silicon rice 2 (Lsi2), which has no similarity to Lsi1. This gene is constitutively expressed in the roots. The protein encoded by this gene is localized, like Lsi1, on the plasma membrane of cells in both the exodermis and the endodermis, but in contrast to Lsi1, which is localized on the distal side, Lsi2 is localized on the proximal side of the same cells. Expression of Lsi2 in Xenopus oocytes did not result in influx transport activity for silicon, but preloading of the oocytes with silicon resulted in a release of silicon, indicating that Lsi2 is a silicon efflux transporter. The identification of this silicon transporter revealed a unique mechanism of nutrient transport in plants: having an influx transporter on one side and an efflux transporter on the other side of the cell to permit the effective transcellular transport of the nutrients.

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  • Characterization of silicon permeability of NIP in gramineous plants Reviewed

    Mitani Namiki, Yamaji Naoki, Katsuhara Maki, Ma Jian Feng

    PLANT AND CELL PHYSIOLOGY   48   S30   2007

  • Gene expression and functional analysis of a candidate gene related to Al-activated citrate transporter in barley Reviewed

    Furukawa Jun, Yamaji Naoki, Wang Hua, Katsuhara Maki, Sato Kazuhiro, Takeda Kazuyoshi, Ma Jian Feng

    Plant and Cell Physiology   48   S44   2007

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  • Low Temperature and Aquaporins, a Molecular Mechanism of Water Transport

    KATSUHARA Maki, CHUNG Gap Chae, SAKURAI Junko, MURAI Mari, IZUMI Yohei, TSUMUKI Hisaaki

    Cryobiology and cryotechnology   53 ( 1 )   21 - 32   2007

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    Aquaporins are membrane proteins facilitating the membrane transport of water and low molecular weight compounds. Recent studies have indicated that aquaporins are involved in cellular tolerance mechanisms under low temperature stress in plants and insects. Critical roles of aquaporins in cold tolerance and the influence of H_2O_2 were suggested in the cucumber and figleaf gourd. Reduction of water uptake was observed in rice under chilling stress, in which the down-regulation of aquaporin expression and probable inactivation of aquaporins were involved. Glycerol and water transport by aquaporins was investigated, and revealed to be essential for freezing tolerance in insect cells.

    DOI: 10.20585/cryobolcryotechnol.53.1_21

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  • Synchronous changes of aquaporins in rice seedlings during early phase of salt-stress.

    Mineo Shibasaka, Maki Katsuhara

    PLANT AND CELL PHYSIOLOGY   48   S162 - S162   2007

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  • Comparison of transpiration rates of naturalized and domestic Sedum species

    Izumi Mori, Takashi Enomoto, Maki Katsuhara

    PLANT AND CELL PHYSIOLOGY   48   S227 - S227   2007

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  • The BnALMT1 Protein That is an Aluminum-Activated Malate Transporter is Localized in the Plasma Membrane. International journal

    Ayalew Ligaba, Maki Katsuhara, Wataru Sakamoto, Hideaki Matsumoto

    Plant signaling & behavior   2 ( 4 )   1 - 2   2007

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    We have previously reported that Al-induces citrate and malate efflux from P-sufficient and P-deficient plants of rape (Brassica napus L.) and that P-deficiency alone could not induce this response. Further investigation showed that the transcript of two genes designated BnALMT1 and BnALMT2 is accumulated in roots by Al-treatment. Transgenic tobacco cells (Nicotiana tabacum) and Xenopus laevis oocytes expressing the BnALMT1 and BnALMT2 proteins released more malate than control cells in the presence of Al, indicating that the BnALMT genes encode an Al-activated malate transporter. The transgenic tobacco cells exposed to toxic level of Al grew better than control cells indicating that the genes can enhance Al-resistance of plant cells. In this study we showed the subcellular localization of BnALMT1 fused to the green fluoresce protein (GFP). The BnALMT1:: GFP construct was transiently expressed in protoplasts prepared from Arabidopsis leaves using the polyethylene glycol (PEG) method. The result showed that the BnALMT1 protein is localized in the plasma membrane. This provides further evidence that the BnALMT proteins facilitate the transport of malate across the plasma membrane (PM).

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  • Molecular mechanisms of water uptake and transport in plant roots: research progress with water channel aquaporins

    Katsuhara Maki

    Plant Root   1   22 - 26   2007

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    Aquaporins (water channels) are membrane proteins which facilitate the transport of water and low molecular weight compounds across biological membranes. The author and collaborators identified barley aquaporins and investigated the level of transcripts in roots, water transport activity, tissue localization, expression reduction by salt stress, and diurnal changes in the expression of a particular aquaporin. Over-expression of a barley aquaporin, HvPIP2;1, increased the shoot/root ratio and raised salt sensitivity in transgenic rice plants. Aquaporin research is providing significant insights into the water relations of plant roots.

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  • The BnALMT1 and BnALMT2 genes from rape encode aluminum-activated malate transporters that enhance the aluminum resistance of plant cells

    Ayalew Ligaba, Maki Katsuhara, Peter R. Ryan, Mineo Shibasaka, Hideaki Matsumoto

    PLANT PHYSIOLOGY   142 ( 3 )   1294 - 1303   2006.11

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    The release of organic anions from roots can protect plants from aluminum (Al) toxicity and help them overcome phosphorus ( P) deficiency. Our previous findings showed that Al treatment induced malate and citrate efflux from rape ( Brassica napus) roots, and that P deficiency did not induce the efflux. Since this response is similar to the malate efflux from wheat ( Triticum aestivum) that is controlled by the TaALMT1 gene, we investigated whether homologs of TaALMT1 are present in rape and whether they are involved in the release of organic anions. We isolated two TaALMT1 homologs from rape designated BnALMT1 and BnALMT2 (B. napus Al-activated malate transporter). The expression of these genes was induced in roots, but not shoots, by Al treatment but P deficiency had no effect. Several other cations ( lanthanum, ytterbium, and erbium) also increased BnALMT1 and BnALMT2 expression in the roots. The function of the BnALMT1 and BnALMT2 proteins was investigated by heterologous expression in cultured tobacco ( Nicotiana tabacum) cells and in Xenopus laevis oocytes. Both transfection systems showed an enhanced capacity for malate efflux but not citrate efflux, when exposed to Al. Smaller malate fluxes were also activated by ytterbium and erbium treatment. Transgenic tobacco cells grew significantly better than control cells following an 18 h treatment with Al, indicating that the expression of BnALMT1 and BnALMT2 increased the resistance of these plant cells to Al stress. This report demonstrates that homologs of the TaALMT1 gene from wheat perform similar functions in other species.

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  • A silicon transporter in rice

    JF Ma, K Tamai, N Yamaji, N Mitani, S Konishi, M Katsuhara, M Ishiguro, Y Murata, M Yano

    NATURE   440 ( 7084 )   688 - 691   2006.3

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    Silicon is beneficial to plant growth and helps plants to overcome abiotic and biotic stresses by preventing lodging ( falling over) and increasing resistance to pests and diseases, as well as other stresses(1-3). Silicon is essential for high and sustainable production of rice(4), but the molecular mechanism responsible for the uptake of silicon is unknown. Here we describe the Low silicon rice 1 (Lsi1) gene, which controls silicon accumulation in rice, a typical silicon-accumulating plant. This gene belongs to the aquaporin family(5) and is constitutively expressed in the roots. Lsi1 is localized on the plasma membrane of the distal side of both exodermis and endodermis cells, where casparian strips are located. Suppression of Lsi1 expression resulted in reduced silicon uptake. Furthermore, expression of Lsi1 in Xenopus oocytes showed transport activity for silicon only. The identification of a silicon transporter provides both an insight into the silicon uptake system in plants, and a new strategy for producing crops with high resistance to multiple stresses by genetic modification of the root's silicon uptake capacity.

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  • Hydraulic conductivity and aquaporins of cortical cells in gravitropically bending roots of Pisum sativum L.

    N Miyamoto, M Katsuhara, T Ookawa, K Kasamo, T Hirasawa

    PLANT PRODUCTION SCIENCE   8 ( 5 )   515 - 524   2005.12

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    We examined the differential elongation of gravitropically bending roots of Pisum sativum L. in terms of cell enlargement and water uptake by cells in the growing tissue. Hydraulic conductivity between the elongating and mature tissues (Lp) was estimated from the equation G = A x Lp x AT, where G is the water-uptake rate, A is the surface area of a single cell and AP is the driving force. The rate of entry of water into a cell was estimated from the rate of increase in the volumes of cells in the outer cortex, which were calculated from longitudinal sections at given times. Gravitropic bending occurred I h after the application of gravi-stimulation and the curvature increased rapidly for the next 3 h. The biggest difference in the partial elongation rate between opposite sides of a root was found in the region 3 to 4 mm from the root tip at the start of stimulation. Cell enlargement rate was 2.8 to 3.8 times greater on the upper side of the root than on the lower side. The water potential and the osmotic potential, in both the elongating and mature tissues, were the same on both sides of the root. Therefore, there was no difference in the driving force for water flow. Hydraulic conductivity was 2.3 to 4.2 times greater on the upper side of the root than on the lower side. There was no difference between the upper and lower sides of the root in the amounts of 19-kD and 24-kD proteins in membrane fractions, which we assumed to be aquaporins (putative aquaporins), as estimated with two preparations of polyclonal antibodies. The differential elongation that Occurred during root gravitropism was caused by a difference in Lp. However, the difference in Lp did not appear to be regulated by the concentration in cell membranes of the putative aquaporins.

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  • Salt stress-induced lipid peroxidation is reduced by glutathione S-transferase, but this reduction of lipid peroxides is not enough for a recovery of root growth in Arabidopsis

    M Katsuhara, T Otsuka, B Ezaki

    PLANT SCIENCE   169 ( 2 )   369 - 373   2005.8

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    Reactive oxygen species (ROS)-related membrane lipid peroxidation in the root of Arabidopsis thaliana was fluorescently visualized and investigated under salt stress. In the control roots without salt stress, more fluorescence was observed in the elongating region than in the meristematic region. Salt stress of 100 mM NaCl enhanced the fluorescence in both, indicating that salt stress-induced ROS, and consequently membrane lipid peroxidation. In transgenic tobacco glutathione S-transferase over-expressing Arabidopsis (the parB plants), less fluorescence was observed than in the non-transgenic control plants. In the salt-stressed parB plant roots, the fluorescent brightness was reduced to 46% of that of the non-transgenic plant in the meristematic region. However, the inhibition of root growth was not improved in parB plants under salt stress at pH 5.7. That is, 100 mM of salt stress reduced the root growth to 40% or less both in the parent control plants and the parB plants. The root tissue osmotic pressure was almost the same between the two tested lines, which may be one of the reasons why no difference was observed in root growth between the two lines. These results suggested that salt stress-induced oxidative stress, and the introduction/overexpression of a glutathione S-transferase gene may have reduced the amount of ROS but the removal of ROS was not sufficient to effect salt tolerance because salt stress also caused an osmotic imbalance reducing the root growth. (c) 2005 Elsevier Ireland Ltd. All rights reserved.

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  • A novel cyanobacterial SmtB/ArsR family repressor regulates the expression of a CPx-ATPase and a metallothionein in response to both Cu(I)/Ag(I) and Zn(II)/Cd(II)

    T Liu, S Nakashima, K Hirose, M Shibasaka, M Katsuhara, B Ezaki, DP Giedroc, K Kasamo

    JOURNAL OF BIOLOGICAL CHEMISTRY   279 ( 17 )   17810 - 17818   2004.4

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    A novel SmtB/ArsR family metalloregulator, denoted BxmR, has been identified and characterized from the cyanobacterium Oscillatoria brevis. Genetic and biochemical evidence reveals that BxmR represses the expression of both bxa1, encoding a CPx-ATPase metal transporter, as well as a divergently transcribed operon encoding bxmR and bmtA, a heavy metal sequestering metallothionein. Derepression of the expression of all three genes is mediated by both monovalent (Ag(I) and Cu(I)) and divalent (Zn(II) and Cd(II)) heavy metal ions, a novel property among SmtB/ArsR metal sensors. Electrophoretic gel mobility shift experiments reveal that apoBxmR forms multiple resolvable complexes with oligonucleotides containing a single 12-2-12 inverted repeat derived from one of the two operator/promoter regions with similar apparent affinities. Preincubation with either monovalent or divalent metal ions induces disassembly of both the BxmR-bxa1 and BxmR-bxmR/bmtA operator/promoter complexes. Interestingly, the temporal regulation of expression of bxa1 and bmtA mRNAs is different in O. brevis with bxa1 induced first upon heavy metal treatment, followed by bmtA/bxmR. A dynamic interplay among Bxa1, BmtA, and BxmR is proposed that maintains metal homeostasis in O. brevis by balancing the relative rates of metal storage and efflux of multiple heavy metal ions.

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  • A wheat gene encoding an aluminum-activated malate transporter

    T Sasaki, Y Yamamoto, B Ezaki, M Katsuhara, SJ Ahn, PR Ryan, E Delhaize, H Matsumoto

    PLANT JOURNAL   37 ( 5 )   645 - 653   2004.3

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    The major constraint to plant growth in acid soils is the presence of toxic aluminum (Al) cations, which inhibit root elongation. The enhanced Al tolerance exhibited by some cultivars of wheat is associated with the Al-dependent efflux of malate from root apices. Malate forms a stable complex with Al that is harmless to plants and, therefore, this efflux of malate forms the basis of a hypothesis to explain Al tolerance in wheat. Here, we report on the cloning of a wheat gene, ALMT1 (aluminum-activated malate transporter), that co-segregates with Al tolerance in F-2 and F-3 populations derived from crosses between near-isogenic wheat lines that differ in Al tolerance. The ALMT1 gene encodes a membrane protein, which is constitutively expressed in the root apices of the Al-tolerant line at greater levels than in the near-isogenic but Al-sensitive line. Heterologous expression of ALMT1 in Xenopus oocytes, rice and cultured tobacco cells conferred an Al-activated malate efflux. Additionally, ALMT1 increased the tolerance of tobacco cells to Al treatment. These findings demonstrate that ALMT1 encodes an Al-activated malate transporter that is capable of conferring Al tolerance to plant cells.

    DOI: 10.1111/j.1365-313X.2003.01991.x

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  • Overexpression of the barley aquaporin HvPIP2;1 increases internal CO2 conductance and CO2 assimillation in the leaves of transgenic rice plants

    YT Hanba, M Shibasaka, Y Hayashi, T Hayakawa, K Kasamo, Terashima, I, M Katsuhara

    PLANT AND CELL PHYSIOLOGY   45 ( 5 )   521 - 529   2004

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    The internal conductance for CO2 diffusion (g(i)) and CO2 assimilation rate were measured and the related anatomical characteristics were investigated in transgenic rice leaves that overexpressed barley aquaporin HvPIP2;1. This study was performed to test the hypothesis that aquaporin facilitates CO2 diffusion within leaves. The g(i) value was estimated for intact leaves by concurrent measurements of gas exchange and carbon isotope ratio. The leaves of the transgenic rice plants that expressed the highest levels of Aq-anti-HvPIP2;1 showed a 40% increase in g(i) as compared to g(i) in the leaves of wild-type rice plants. The increase in g(i) was accompanied by a 14% increase in CO2 assimilation rate and a 27% increase in stomatal conductance (g(s)). The transgenic plants that had low levels of Aq-anti-HvPIP2;1 showed decreases in g(i) and CO2 assimilation rate. In the plants with high levels of Aq-anti-HvPIP2;1, mesophyll cell size decreased and the cell walls of the epidermis and mesophyll cells thickened, indicating that the leaves had become xeromorphic. Although such anatomical changes could partially offset the increase in g(i) by the aquaporin, the increase in aquaporin content overcame such adverse effects.

    DOI: 10.1093/pcp/pch070

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  • Changes in roots and salt sensitivity of rice plants over-expressing aquaporin

    M Katsuhara, K Koshio, M Shibasaka, Y Hayashi, T Hayakawa, K Kasamo

    PLANT AND CELL PHYSIOLOGY   45   S174 - S174   2004

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  • Expression of an aquaporin at night in relation to the growth and root water permeability in barley seedlings

    M Katsuhara, K Koshio, M Shibasaka, K Kasamo

    SOIL SCIENCE AND PLANT NUTRITION   49 ( 6 )   883 - 888   2003.12

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    Both root and shoot of barley seedlings showed continuous growth at night as well as in the daytime. Root hydraulic conductivity (Lp(r)) was monitored, and high Lp(r) values were recorded at night. For revealing the molecular mechanism, the presence of an aquaporin HvPIP2;1 in barley roots was investigated. The accumulation of the HvPIP2;1 transcript was mostly observed from the evening to the middle of the night. HvPIP2;1 protein was most abundant around midnight. As for tissue localization, HvPIP2;1 protein was abundant in cells involved in water transport. These results suggested the possible involvement of HvPIP2;1 in the mechanism of water transport in roots.

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  • Over-expression of a barley aquaporin increased the shoot/root ratio and raised salt sensitivity in transgenic rice plants

    M Katsuhara, K Koshio, M Shibasaka, Y Hayashi, T Hayakawa, K Kasamo

    PLANT AND CELL PHYSIOLOGY   44 ( 12 )   1378 - 1383   2003.12

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    Barley HvPIP2;1 is a plasma membrane aquaporin and its expression was down-regulated after salt stress in barley [Katsuhara et al. (2002) Plant Cell Physiol. 43: 885]. We produced and analyzed transgenic rice plants over-expressing barley HvPIP2;1 in the present study. Overexpression of HvPIP2;1 increased (1) radial hydraulic conductivity of roots (Lp(r)) to 140%, and (2) the mass ratio of shoot to root up to 150%. In these transgenic rice plants under salt stress of 100 mM NaCl, growth reduction was greater than in non-transgenic plants. A decrease in shoot water content (from 79% to 61%) and reduction of root mass or shoot mass (both less than 40% of non-stressed plants) were observed in transgenic plants under salt stress for 2 weeks. These results indicated that over-expression of HvPIP2;1 makes rice plants sensitive to 100 mM NaCl. The possible involvement of aquaporins in salt tolerance is discussed.

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  • A metallothionein and CPx-ATPase handle heavy-metal tolerance in the filamentous cyanobacterium Oscillatoria brevis

    T Liu, S Nakashima, K Hirose, Y Uemura, M Shibasaka, M Katsuhara, K Kasamo

    FEBS LETTERS   542 ( 1-3 )   159 - 163   2003.5

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    A metallothionein (BmtA) and a CPx-ATPase (Bxa1) have been identified and characterized from the cyanobacterium Oscillatoria brevis. Both bmtA and bxa1 expression can be markedly induced in vivo by Zn2+ or Cd2+. Over-expression of bmaA or bxa1 in Escherichia coli enhances Zn2+ and Cd2+ tolerance in the transformant. Dynamic studies on the expression of two genes showed that the maximum expression of bxa1 induced by Zn2+ and Cd2+ was much quicker than that of bmtA, suggesting distinct physiological roles of metallothionein and CPx-ATPase in the handling of surplus metal. (C) 2003 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

    DOI: 10.1016/S0014-5793(03)00370-3

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  • Increase in CO2 permeability (diffusion conductance) in leaves of transgenic rice plant over-expressing barley aquaporin

    M Katsuhara, Y Hanba, M Shibasaka, Y Hayashi, T Hayakawa, K Kasamo

    PLANT AND CELL PHYSIOLOGY   44   S86 - S86   2003

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  • A novel histidine-rich CPx-ATPase from the filamentous cyanobacterium Oscillatoria brevis related to multiple-heavy-metal cotolerance

    L Tong, S Nakashima, M Shibasaka, M Katsuhara, K Kasamo

    JOURNAL OF BACTERIOLOGY   184 ( 18 )   5027 - 5035   2002.9

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    A novel gene related to heavy-metal transport was cloned and identified from the filamentous cyanobacterium Oscillatoria brevis. Sequence analysis of the gene (the Bxa1 gene) showed that its product possessed high homology with heavy-metal transport CPx-ATPases. The CPC motif, which is proposed to form putative cation transduction channel, was found in the sixth transmembrane helix. However, instead of the CXXC motif that is present in the N termini of most metal transport CPx-ATPases, Bxa1 contains a unique Cys-Cys (CC) sequence element and histidine-rich motifs as a putative metal binding site. Northern blotting and real-time quantitative reverse transcription-PCR showed that expression of Bxa1 mRNA was induced in vivo by both monovalent (Cu+ and Ag+) and divalent (Zn2+ and Cd2+) heavy-metal ions at similar levels. Experiments on heavy-metal tolerance in Escherichia coli with recombinant Bxa1 demonstrated that Bxa1 conferred resistance to both monovalent and divalent heavy metals. This is the first report of a CPx-ATPase responsive to both monovalent and divalent heavy metals.

    DOI: 10.1128/JB.184.18.5027-5035.2002

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  • Functional analysis of water channels in barley roots

    M Katsuhara, Y Akiyama, K Koshio, M Shibasaka, K Kasamo

    PLANT AND CELL PHYSIOLOGY   43 ( 8 )   885 - 893   2002.8

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    We identified three genes homologous to water channels in the plasma membrane type subfamily from roots of barley seedlings. These genes were designated HvPIP2;1, HvPIP1;3, and HvPIP1;5 after comparison to Arabidopsis aquaporins. Competitive reverse transcription (RT)-PCR was applied in order to distinguish and to quantify their transcripts. The HvPIP2;1 transcript was the most abundant among the three in roots. Salt stress (200 mM NaCl) down-regulated HvPIP2;1 (transcript and protein), but had almost no effect on the expressions of HvPIP1;3, or HvPIP1;5. Approximately equal amounts of the transcripts of the three were detected in shoots, and salt stress enhanced the expression of HvPIP2;1 but not of HvPIP1;3, or HvPIP1;5. HvPIP2;1 protein was confirmed to be localized in the plasma membrane. Functional expression of HvPIP2;1 in Xenopus oocytes confirmed that HvPIP2;1 encoded an aquaporin that transports water. This water permeability was reduced by HgCl2, which is a typical water channel inhibitor. This activity was not modified by some inhibitors against protein kinase and protein phosphatase.

    DOI: 10.1093/pcp/pcf102

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  • Different mechanisms of four aluminum (Al)-resistant transgenes for Al toxicity in Arabidopsis

    B Ezaki, M Katsuhara, M Kawamura, H Matsumoto

    PLANT PHYSIOLOGY   127 ( 3 )   918 - 927   2001.11

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    We have characterized the mechanism of action of four transgenes (AtBCB [Arabidopsis blue copper-binding protein], parB [tobacco {Nicotiana tabacum} glutathione S-transferase], NtPox [tobacco peroxidase], and NtGDI1 [tobacco GDP dissociation inhibitor]) that independently Al resistance on transgenic Arabidopsis. All four transgenic lines showed lower deposition of callose after Al treatment than the Landsberg erecta ecotype of Arabidopsis, confirming that the four genes function to ameliorate Al toxicity. Influx and efflux experiments of Al ions suggested that the AtBCB gene may suppress Al absorption, whereas expression of the NtGDI1 gene promotes a release of Al in the root tip region of Arabidopsis. The total enzyme activities of glutathione S-transferases or peroxidases in transoenic lines carrying either the parB or NtPox genes were significantly higher than in the Landsberg erecta ecotype of Arabidopsis, and these enzyme activities were maintained at higher levels during Al stress. Furthermore, lipid peroxidation caused by Al stress was repressed in these two transgenic lines, suggesting that overexpression of these two genes diminishes oxidative damage caused by Al stress. Al-treated roots of trans-enic plants were also stained by 4',6-diamino-2-phenylindole to monitor cell death caused by Al toxicity. The result suggested that cell death is repressed in the NtPox line. Analysis of F-1 hybrids between the four transgenic lines suggests that more resistant transgenic plants can be constructed by combinations of these four genes.

    DOI: 10.1104/pp.127.3.918

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  • Isolation of barley saLT gene: Its relation to salt tolerance and to hormonal regulation by abscisic acid and jasmonic acid

    M Katsuhara, M Yamada, K Kasamo

    SOIL SCIENCE AND PLANT NUTRITION   47 ( 1 )   187 - 193   2001.3

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    A homologous gene to rice salT was isolated from barley and designated as Hv-salT, Since the expression of rice salT was regulated by plant hormones (abscisic acid and jasmonic acid) and salt stress, the effects of these plant hormones and salt stress on the growth of barley seedlings and the expression of Hv-salT were investigated. In contrast to rice salT, almost no changes in the expression of Hv-salT by hormones and salt stress were detected.

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  • Expression and stress-dependent induction of potassium channel transcripts in the common ice plant

    H Su, D Golldack, M Katsuhara, CS Zhao, HJ Bohnert

    PLANT PHYSIOLOGY   125 ( 2 )   604 - 614   2001.2

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    We have characterized transcripts for three potassium channel homologs in the AKT/KAT subfamily (Shaker type) from the common ice plant (Mesembryanthemum crystallinum), with a focus on their expression during salt stress (up to 500 mM NaCl). Mkt1 and 2, Arabidopsis AKT homologs, and Kmt1, a KAT homolog, are members of small gene families with two to three isoforms each. Mkt1 is root specific; Mkt2 is found in leaves, flowers, and seed capsules; and Kmt1 is expressed in leaves and seed capsules. Mkt1 is present in all cells of the root, and in leaves a highly conserved isoform is detected present in all cells with highest abundance in the vasculature. MKT1 for which antibodies were made is localized to the plasma membrane. Following salt stress, MKT1 (transcripts and protein) is drastically down-regulated, Mkt2 transcripts do not change significantly, and Kmt1 is strongly and transiently (maximum at 6 h) up-regulated in leaves and stems. The detection and stress-dependent behavior of abundant transcripts representing subfamilies of potassium channels provides information about tissue specificity and the complex regulation of genes encoding potassium uptake systems in a halophytic plant.

    DOI: 10.1104/pp.125.2.604

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  • Identification and characterization of a putative heavy metal transport gene from a cyanobacterium Oscillatoria Brevis :

    Liu Tong, NAKASHIMA Susumu, SHIBASAKA Mineo, KATSUHARA Maki, KASAMO Kunihiro

    Plant and cell physiology   42   s208   2001

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  • In situ RNA hybridization using Technovit resin in Arabidopsis thaliana

    K Takechi, W Sakamoto, M Katsuhara, M Murata, F Motoyoshi

    PLANT MOLECULAR BIOLOGY REPORTER   17 ( 1 )   43 - 51   1999

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    A protocol is described for RNA in situ hybridization using thin sections prepared by Technovit resin. Technovit is a widely used resin for histological examinations. Since it does not require time-consuming processes such as removal of the resin and can be performed without high temperature treatment, a high resolution of sections could be possible compared to other resins and paraffin. Thin sections (approximately 4 mu m) were made from inflorescences of Arabidopsis thaliana embedded in Technovit 8100 resin, and in situ hybridization was performed using the protocol described in this article. Hybridization signals were observed using LEAFY and other genes as probes, showing that this resin can be used for in situ analysis. In our experiments, the most important factor for a successful in situ, hybridization pattern was to optimize the RNase A concentration after hybridization. We routinely used RNase A at a concentration of 2-5 ng/ml, a concentration much lower than that used for paraffin embedding method. Thus, the use of the Technovit resin for plant tissue embedding results in a faster protocol and greater quality than allowed by paraffin sections.

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  • Intracellular pH and proton-transport in barley root cells under salt stress: In vivo 31P-NMR study

    Maki Katsuhara, Yoshiaki Yazaki, Katsuhiro Sakano, Toshio Kawasaki

    Plant and Cell Physiology   38 ( 2 )   155 - 160   1997

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    Salt stress-induced changes of intracellular pH and in levels of phosphorous compounds were monitored in intact root tips of barley seedlings (Hordeum vulgare cv. Akashinriki) by in vivo 31P-nuclear magnetic resonance (NMR) spectroscopy. Vacuolar alkalization was observed after treatment with both 300 and 500 mM NaCl. Much of the observed apparent alkalization of the cytoplasm was eliminated when the effect of Na+ ions on the titration curve was considered. Within 1 h after the initiation of salt stress, levels of glucose-6-phosphate and UDP-glucose decreased markedly, and such decreases might lead directly or indirectly to cell death. Simultaneous measurements of the external and intracellular pH revealed the promotion of external acidification and internal alkalization during salt stress. Possible mechanisms of Na+/H+ antiport at the tonoplast and the role of proton-pump in the plasma membrane are discussed.

    DOI: 10.1093/oxfordjournals.pcp.a029146

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  • Salt stress induced nuclear and DNA degradation in meristematic cells of barley roots

    Maki Katsuhara, Toshio Kawasaki

    Plant and Cell Physiology   37 ( 2 )   169 - 173   1996

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    Root growth of barley (Hordeum vulgare L., ev. Akashinriki) was inhibited by 200 mM NaCl, when 1 mM CaCl2 was present in the hydroponic culture solution. Increasing the CaCl2 up to 10 mM partially prevented this inhibition. However, inhibition also occurred with 100 mM NaCl in the presence of 0.1 mM CaCl2. The nuclei of meristematic cells in roots in which growth had been inhibited by salt stress were studied after staining with DAPI (4',6-diamino-2-phenylindol). Nuclear deformation of the cells occurred with 12 h of salt stress with 500 mM NaCl, and was followed by degradation. The nuclear degradation was also observed when the roots were exposed to more than 300 mM NaCl for 24 h. Biochemical analysis revealed that nuclear degradation was accompanied by apoptosislike DNA fragmentation. The intracellular mechanisms of nuclear degradation in cells after salt stress are discussed.

    DOI: 10.1093/oxfordjournals.pcp.a028928

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  • Distribution and Production of trans-Aconitic Acid in Barnyard Grass (Echinochloa crus-galli var. oryzicola) as Putative Antifeedant against Brown Planthoppers :

    KATSUHARA Maki, SAKANO Katsuhiro, SATO Mamoru, KAWAKITA Hiromu, KAWABE Susumu

    Plant and cell physiology   34 ( 2 )   251 - 254   1993

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    A quantitative analysis of organic acid in leaves of barnyard grass revealed that the contents of trans-aconitic acid were high, suggesting that this compound may act as an antifeedant against brown planthoppers. However, trans-aconitic acid could not be detected in the phloem sap which was considered to be the main nutrient source for brown planthoppers. trans-Aconitic acid was formed in vitro from cis-aconitic acid through the aconitate isomerase activity which was detected only in the leaf sheath, but not in the leaf blade.

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    Other Link: https://projects.repo.nii.ac.jp/?action=repository_uri&item_id=181825

  • Detection of mycoplasmalike organisms in the phloem sap collected from mulberry trees with dwarf disease.

    KAWAKITA HIROMU, KATSUHARA MAKI

    Journal of Insect Biotechnology and Sericology   62 ( 2 )   125 - 131   1993

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    Mycoplasmalike organisms (MLOs), the pathogens of mulberry dwarf disease, were succesufully isolated by stylectomy method (cutting of insect stylets by YAG laser) using a homopterous insect, Ricania japonica. The MLOs were detectable in the phloem sap collected from diseased mulberry plants as fluorescent particles stained with 4, 6-diamidino-2-phenylindole (DAPI) under a fluorescence microscope but were not detected in the sap of healthy mulberry plants. The presence of MLOs in the phloem sap was also confirmed by electronmicroscopy.<br>On the other hand, two kinds of leafhoppers, vectors of mulberry dwarf MLOs, and other some tested hemipterous insects could not used for this laser stylectomy. Scanning-electronmicroscopical observation showed that the stylets of these leafhoppers which were not suitable for the stylectomy method were pushed out or broken after laser cutting, whwereas stylets of Ricania japonica remained firmly in the tissues.

    DOI: 10.11416/kontyushigen1930.62.125

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    Other Link: http://agriknowledge.affrc.go.jp/RN/2010510238

  • Cytoplasmic Alkalization and Cytoplasmic Streaming Induced by Light and Histidine in Leaf Cells of Egeria densa: in vivo 31P-NMR study

    Yoshito Tominaga, Kazuyuki Kuchitsu, Maki Katsuhara, Masashi Tazawa, Shigetoh Miyachi

    Plant and Cell Physiology   32 ( 2 )   261 - 268   1991.3

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  • Mechanism of calcium‐dependent salt tolerance in cells of Nitellopsis obtusa: role of intracellular adenine nucleotides

    M. KATSUHARA, M. TAZAWA

    Plant, Cell &amp; Environment   13 ( 2 )   179 - 184   1990

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    Abstract. The tonoplasts of internodal cells of Nitellopsis were removed by perfusing the vacuoles with media containing a Ca2 chelator, EGTA. Treatment of tonoplast‐free cells with 100 mol m3 NaCl induces a large membrane depolarization, a drastic decrease in the membrane resistance and an increase in Na+ influx. These events are identical to those that occur in intact cells subjected to high NaCl. These responses to NaCl are prevented if 10 mol m3 Ca2+ is supplied together with 100 mol m3 NaCl. The protective effect of Ca2+ is evident only when the intracellular ATP concentration exceeds 0.1 mol m3 and does not occur full when the intracellular ATP is removed. AMP at concentrations greater than 0.5 mol m3 or 0.25 mol m3 AMPPNP can replace ATP. It is concluded that ATP does not act as an energy source nor as a substrate for protein phosphorylation. ATP seems to exert its effects as a coeffector with Ca2+ in regulating the Na+ permeability of the plasma membrane. Copyright © 1990, Wiley Blackwell. All rights reserved

    DOI: 10.1111/j.1365-3040.1990.tb01290.x

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  • ATP-regulated ion channels in the plasma membrane of a characeae alga, Nitellopsis obtusa

    Maki Katsuhara, Tetsuro Mimura, Masashi Tazawa

    Plant Physiology   93 ( 1 )   343 - 346   1990

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    Using the patch-clamp technique, we recorded single-channel currents across the excised patch of the plasma membrane of Nitellopsis. Both K+ and Na+ can pass this channel, but currents were not carried by Cl-. Upon the addition of ATP or AMP to the cytoplasmic side, the frequency of channel opening decreased. This is the first report on an ATP-regulated channel in plant cells.

    DOI: 10.1104/pp.93.1.343

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  • Patch-clamp study on a Ca2+-regulated K+ channel in the tonoplast of the brackish characeae lamprothamnium succinctum

    Maki Katsuhara, Tetsuro Mimura, Masashi Tazawa

    Plant and Cell Physiology   30 ( 4 )   549 - 555   1989

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    Cytoplasmic drops were prepared from internodal cells of the brackish Characeae Lamprothamnium succinctum. Applying the patch-clamp technique to single drops covered with tonoplast, we demonstrated the presence of Ca2+-regulated K+ channels in the tonoplast. In a cell-attached mode, the selectivity of such channels for K+ was about 50 times that for Na+. This channel showed a tendency to rectify in an outward direction. In the negative region of the pipette voltage, the conductance of this channel was 50 pS, while it was 100 pS in the positive voltage region. When the pipette voltage was increased above 50 mV, two conductance levels were found in the cell-attached mode as well as in the excised patch (cytoplasmic-side-out patch), which was obtained by pulling the patch pipette from the cytoplasmic drop under conditions of low levels of Ca2+. Using the excised patch, we controlled the level of Ca2+ on the cytoplasmic side of the channels. At a low level of Ca2+ (pCa=8) on the cytoplasmic side, the open frequency was very low and the opening time was short. An increase in Ca2+ on the cytoplasmic side (pCa = 5) increased both the frequency and the duration of opening. However, the conductance of the channels did not change. This regulation by Ca2+ of the K+ channels was reversible, that is, addition of EGTA on the cytoplasmic side inactivated the channels. The present study demonstrates a direct action of Ca2+ on the K+ channels. The physiological role of the K+ channel in the regulation of turgor in Lamprothamnium is discussed. © 1989 The Japanese Society of Plant Physiologists (JSPP).

    DOI: 10.1093/oxfordjournals.pcp.a077774

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  • Rapid cytoplasmic alkalization and dynamics of intracellular compartmentation of inorganic phosphate during adaptation against salt stress in a halotolerant unicellular green alga dunaliella tertiolecta:31P-nuclear magnetic resonance study Reviewed

    Kuchitsu, K., Katsuhara, M., Miyachi, S.

    Plant and Cell Physiology   30 ( 3 )   407 - 414   1989

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  • ATP IS ESSENTIAL FOR CALCIUM-INDUCED SALT TOLERANCE IN NITELLOPSIS-OBTUSA

    M KATSUHARA, M TAZAWA

    PROTOPLASMA   138 ( 2-3 )   190 - 192   1987

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Books

  • 根っこってなんだろう?

    且原真木, 松波麻耶( Role: Joint author ,  根っこはどうやって水を吸収するの)

    文研出版  2023.8  ( ISBN:9784580825970

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    Total pages:47p   Responsible for pages:10-11p   Language:Japanese Book type:Textbook, survey, introduction

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  • バイオスティミュラントハンドブック : 植物の生理活性プロセスから資材開発、適用事例まで

    日本バイオスティミュラント協議会, 山内, 靖雄, 須藤, 修, 和田, 哲夫( Role: Contributor ,  第1編第2章第4節 水ストレスが植物に与える影響と植物の応答)

    エヌ・ティー・エス  2022.4  ( ISBN:9784860437695

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    Total pages:iii, xiii, 460, xiip, 図版vip   Language:Japanese

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  • 植物学の百科事典(共著)

    丸善  2016 

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  • 植物栄養学第2版(共著)

    文永堂  2010 

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  • 水とからだの事典(共著)

    朝倉書店  2008 

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  • みずみずしい体のしくみ:水の通り道「アクアポリン」の働きと病気」

    クバプロ  2005 

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  • 植物の膜輸送システム ポンプ・トランスポーター・チャンネル研究の新展開

    秀潤社  2003 

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  • Molecular aspects of potassium and water transport, and their bearing under salt stress

    Plant Nutrition for sustainable food production and environment 

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  • パッチクランプによるチャンネルの解析

    「植物の細胞を観る実験プロトコール」 

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  • Molecular aspects of potassium and water transport, and their bearing under salt stress

    Plant Nutrition for sustainable food production and environment 

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MISC

  • Analysis of CO2 transport activity of plasma membrane-associated aquaporins of tomato

    光本晏理, 宇都木繁子, 土屋善幸, 且原真木, 森泉

    日本植物生理学会年会(Web)   64th   2023

  • Controls of the water transport activities of Arabidopsis tonoplast intrinsic proteins 3, AtTIP3s and the effects on seed development and germination

    宇都木繁子, 且原真木

    日本植物生理学会年会(Web)   64th   2023

  • Controls of the water transport activities of Arabidopsis tonoplast intrinsic proteins 3, AtTIP3s

    宇都木繁子, 且原真木

    日本植物生理学会年会(Web)   63rd   2022

  • 植物のアクアポリン

    且原 真木

    脳神経内科   94 ( 5 )   635 - 642   2021.5

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  • オオムギ種子における液胞膜型アクアポリン(HvTIPs)の役割

    宇都木繁子, 且原真木

    日本植物生理学会年会(Web)   60th   2019

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  • 種子で発現するオオムギ液胞膜型アクアポリン(HvTIPs)のタンパク質間相互作用

    宇都木繁子, 柴坂三根夫, 且原真木

    日本植物生理学会年会(Web)   59th   2018

  • オオムギ種子における液胞膜型アクアポリンの水輸送活性の調節

    宇都木繁子, 柴坂三根夫, 且原真木

    日本植物生理学会年会要旨集   55th   2014

  • 種子特異的に発現するオオムギTIP3;1による水輸送活性の調節

    宇都木繁子, 篠野静香, 柴坂三根夫, 且原真木

    日本植物生理学会年会要旨集   54th   2013

  • 原形質膜型アクアポリンPIP1とPIP2の共発現による活性抑制

    柴坂三根夫, 篠野静香, 宇都木繁子, 且原真木

    日本植物生理学会年会要旨集   54th   2013

  • オオムギMIPファミリーの解析

    宇都木繁子, 柴坂三根夫, 且原真木, 前川雅彦

    育種学研究   13   2011

  • Varietal differences in physiological responses to salt stress in barley seedings

    YAMAGUCHI Makoto, WATANABE Tamaki, KATSUHARA Maki, TAKEDA Kazuyoshi, OOKAWA Taiichiro, HIRASAWA Tadashi

    79   182 - 183   2010.9

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  • 岡山大学資源生物科学研究所における屋上緑化による建物冷却効果

    且原真木, 田中丸重美, 森泉, 谷明生, 宇都木繁子, 榎本敬, 米谷俊彦

    環境制御   31 ( 1 )   21 - 25   2009

  • 新しい緑化ブロック作成技術

    且原真木

    環境浄化技術   8 ( 3 )   32 - 36   2009

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  • コムギ種子特異的な液胞型アクアポリン(TIP)の解析

    宇都木繁子, 且原真木, 前川雅彦

    育種学研究   11   2009

  • 105 Varietal differences in root hydraulic conductivity and the rate of leaf photosyntesis under salt stress conditions in barley seedlings

    Watanabe Tamaki, Sato Kosuke, Katsuhara Maki, Ookawa Taiichiro, Takeda Kazuyoshi, Hirasawa Tadashi

    Japanese Journal of Crop Science   77 ( 2 )   210 - 211   2008.9

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  • 9-6 オオムギにおけるアルミニウムによるクエン酸分泌に関与する遺伝子の単離と機能解析(9.植物の無機栄養,2007年度東京大会)

    古川 純, 山地 直樹, 王 華, 三谷 奈見季, 村田 佳子, 佐藤 和広, 且原 真木, 武田 和義, 馬 建鋒

    日本土壌肥料学会講演要旨集   ( 53 )   64 - 64   2007.8

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  • B201 アズキゾウムシにおけるオス由来のメスの交尾抑制物質の生化学的分析

    山根 隆史, 宮竹 貴久, 木村 吉伸, 村田 芳行, 且原 真木, 中島 修平

    日本応用動物昆虫学会大会講演要旨   ( 51 )   20 - 20   2007.3

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  • オオムギのアルミニウム活性型クエン酸輸送体候補遺伝子の発現ならびに機能解析

    古川純, 山地直樹, 王華, 且原真木, 佐藤和広, 武田和義, 馬建鋒

    日本植物生理学会年会要旨集   48th   2007

  • イネ科NIPのケイ酸輸送特性の解析

    三谷奈見季, 山地直樹, 且原真木, 馬建鋒

    日本植物生理学会年会要旨集   48th   2007

  • 9-66 イネケイ酸吸収遺伝子Lsi1の輸送特性の解析(9. 植物の無機栄養, 2006年度秋田大会講演要旨)

    三谷 奈見季, 且原 真木, 馬 建鋒

    日本土壌肥料学会講演要旨集   ( 52 )   72 - 72   2006.9

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  • オオムギアクアポリンHvPIP2;1の過剰発現による形質転換イネの葉内CO2拡散コンダクタンスと光合成速度の増加

    半場祐子, 柴坂三根夫, 林泰行, 早川孝彦, 笠毛邦宏, 寺島一郎, 且原真木

    日本植物生理学会年会要旨集   47th   2006

  • アクアポリンの多様性と多面性:耐塩性や光合成との関係

    且原真木, 柴坂三根夫, 半場祐子

    日本植物生理学会年会要旨集   46th   2005

  • 植物の根に関する諸問題-水透過性の分子機構:根における水チャネル・アクアポリンの機能-

    且原真木

    農業および園芸   79:600-605 ( 5 )   600 - 605   2004

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  • A Molecular mechanisms of water absorption and water transport : recent progress in study on water channels

    KATSUHARA Maki

    Root Research   13:15-20 ( 1 )   15 - 20   2004

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    Discovery of aquaporin (water channel) genes have facilitated investigations into the molecular mechanisms of cellular water permeability and transport. We isolated genes of plasma-membrane type aquaporins in barley roots and demonstrated the water transport activity, tissue localization, and diurnal change in expression of a particular aquaporin, HvPIP2;1, in barley. Over-expression of HvPIP2;1 increased the shoot/root ratio and raised salt sensitivity in transgenic rice plants. Furthermore, we noticed the increase in internal CO2 conductance of transgenic plants. This finding indicated that HvPIP2;1 can facilitate permeation of a CO2 as well as water molecule.

    DOI: 10.3117/rootres.13.15

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  • 9-4 コムギのアルミニウム活性化型リンゴ酸トランスポーター遺伝子の機能解析 : 対立遺伝子間の機能相違(9.植物の無機栄養)

    佐々木 孝行, 山本 洋子, 且原 真木, 松本 英明

    日本土壌肥料学会講演要旨集   ( 49 )   66 - 66   2003.8

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  • コムギのA1耐性機構 : リンゴ酸分泌とそのトランスポーター遺伝子

    佐々木 孝行, 山本 洋子, 且原 真木, 大沢 裕樹, 松本 秀明

    根の研究 = Root research   12 ( 2 )   71 - 71   2003.6

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  • A gene encoding an aluminum-activated malate transporter segregates with aluminum tolerance in wheat

    T Sasaki, Y Yamamoto, B Ezaki, M Katsuhara, PR Ryan, E Delhaize, H Matsumoto

    PLANT AND CELL PHYSIOLOGY   44   S84 - S84   2003

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  • Studies on salt stress-tolerant mechanism in barley.1. Analysis of genes expressed specifically under salt stress in salt stress-tolerant barley

    SUGIMOTO M, KATSUHARA M, OAKADA Y, SATO K, ITO K, TAKEDA K

    育種学研究 = Breeding research   4 ( 1 )   20 - 20   2002.3

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  • 9-3 アルミニウム耐性小麦に特異的な遺伝子の機能解析(9.植物の無機栄養)

    佐々木 孝行, 江崎 文一, 山本 洋子, 且原 真木, 松本 英明

    日本土壌肥料学会講演要旨集   ( 48 )   46 - 46   2002.3

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  • Cloning and characterization of a gene expressed in aluminum-tolerant wheat

    T Sasaki, Y Yamamoto, B Ezaki, M Katsuhara, H Matsumoto

    PLANT AND CELL PHYSIOLOGY   43   S51 - S51   2002

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  • 研究室・研究所めぐり(22)岡山大学・資源生物科学研究所

    且原真木

    遺伝   54 ( 2 )   67 - 69   2000

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Presentations

  • Plant molecular physiology of salt stress and salt tolerance mechanisms Invited International conference

    Maki Katsuhara

    The 6th IBOC International Biology Conference 2023  2023.10.25 

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  • Na ion and K ion transport mechanism of ion channel aquaporins. International conference

    Ono S, Katsuhara M

    Taiwan Japan Plant Biology 2023  2023.10.14 

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    Event date: 2023.10.13 - 2023.10.15

    Language:English   Presentation type:Poster presentation  

    Venue:Taipei  

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  • Carbon dioxide transport by Arabidopsis PIP2 aquaporins. International conference

    Tania SS, Utsugi S, Tsuchiya Y, Sasano S, Katsuhara M, Mori I

    Taiwan Japan Plant Biology 2023  2023.10.14 

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    Event date: 2023.10.13 - 2023.10.15

    Language:English   Presentation type:Poster presentation  

    Venue:Taipei  

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  • Identification, characterization, and molecular mechanisms of ion-channel aquaporins in plants. International coauthorship International conference

    Katsuhara M, Tran THS, Ono S, Horie Tomoaki

    Taiwan Japan Plant Biology 2023  2023.10.14 

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    Event date: 2023.10.13 - 2023.10.15

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    Venue:Taipei  

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  • Salt-tolerant Pokkali Rice OsHKT1;1 Variants: Expression and Ion Transport Activity. International conference

    Imran S, Ono S, Tomoaki H, Katsuhara M

    Taiwan Japan Plant Biology 2023  2023.10.14 

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    Event date: 2023.10.13 - 2023.10.15

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    Venue:Taipei  

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  • 根圏でのH2O2とFeによるFenton 反応を制御して、有機質資源利用と地球温暖化阻止を進める

    我妻忠雄, Toan Nguyen Syl, 程 為国, 田原 恒, 且原真木, 土屋善幸, 俵谷圭太郎

    日本土壌肥料学会2023年度愛媛大会  2023.9.13 

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    Event date: 2023.9.12 - 2023.9.14

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:松山  

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  • 植物のイオン輸送性アクアポリン

    且原真木, Tran THS, 小野修太朗, 堀江智明

    第17回トランスポーター研究会  2023.5.27 

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    Event date: 2023.5.27

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:名古屋  

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  • 耐塩性オオムギの根水透過性 (Lpr) 下方制御に関わるアクアポリンHvPIP2;1のPhos-tag® SDS-PAGEを使ったリン酸化解

    大西亜耶, 且原真木

    第17回トランスポーター研究会  2023.5.27 

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    Venue:名古屋  

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  • イオン透過性アクアポリンの Na+イオンおよび K+イオン輸送メカニズム

    小野峻太郎, 且原真木

    第64回日本植物生理学会  2023.3.13 

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    Event date: 2023.3.13 - 2023.3.17

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    Venue:仙台  

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  • シロイヌナズナ液胞膜型アクアポリン AtTIP3 サブファミリーの水輸送活性調節と種子発達と発芽への影響

    宇都木繁子, 且原真木

    第64回日本植物生理学会  2023.3.13 

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    Event date: 2023.3.13 - 2023.3.17

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    Venue:仙台  

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  • トマト原形質膜局在型アクアポリンの CO2 輸送活性の解析

    光本晏理, 宇都木繁子, 土屋善幸, 且原真木, 森泉

    第64回日本植物生理学会  2023.3.13 

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    Venue:仙台  

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  • サボテンの高温・乾燥耐性を支えるアクアポリンの組織分布と生理機能の解明

    佐藤良介, 榊原光, 水口慎太郎, 御堂育子, 鈴木孝征, 堀部貴紀, 柘植尚志, 且原真木, 前島正義

    第64回日本植物生理学会  2023.3.13 

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    Venue:仙台  

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  • 塩ストレス初期段階における耐塩性オオムギK305の根水透過性 (Lpr) 下方制御とオオムギアクアポリンHvPIP2;1のリン酸化の解析

    大西亜耶, 且原真木

    第56回根研究集会(福岡)(オンライン)  2022.9.17 

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    Event date: 2022.9.17 - 2022.9.18

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  • イネにおけるイオン輸送性アクアポリンの同定と機能解析

    且原真木, Sen Tran, 堀江智明

    日本植物学会第86回大会(京都)  2022.9.17 

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    Event date: 2022.9.15 - 2022.9.19

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  • 鉄との協同による植物の新規な根圏改変機能 - 根圏フェントン反応による有機環境の改変・ストレスの回避 ①

    我妻忠雄, 田原恒, 且原真木, 土屋善幸, 俵谷圭太郎

    日本土壌肥料学会2022年度大会  2022.9 

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    Event date: 2022.9.13 - 2022.9.15

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  • サボテンの高い環境耐性に対するアクアポリンの生理的役割の解明

    佐藤 良介, 鈴木孝征, 朝比奈 雅志, 堀部 貴紀, 柘植尚志, 且原真木, 前島 正義

    第63回日本植物生理学会(つくば)(オンライン)  2022.3 

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  • シロイヌナズナ液胞膜型アクアポリンAtTIP3ファミリーの水輸送活性の制御

    宇都木 繁子, 且原 真木

    第63回日本植物生理学会(つくば)(オンライン)  2022.3 

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    Event date: 2022.3.22 - 2022.3.24

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  • シロイヌナズナ液胞膜型アクアポリンAtTIP3 の水輸送活性をコントロールするC末端領域の解析

    宇都木 繁子, 且原 真木

    第62回日本植物生理学会(島根)(オンライン)  2021.3 

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  • イネアクアポリンOsPIP2;4 の発現量と根水透過性(Lpr)の相関性

    大西 亜耶, 且原 真木

    第62回日本植物生理学会(島根)(オンライン)  2021.3 

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  • Ca2+-sensitive and non-selective Na+/K+ channel activity of a barley aquaporin HvPIP2;8.

    Tran S, Horie T, Imran S, Qiu J, McGaughey S, Byrt SC, Tyerman SD, Katsuhara M

    2021.3 

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  • シロイヌナズナの液胞膜型アクアポリンTIP2;2 の機能解析

    山成 由佳, 中原 由揮, 藤本 緋菜, 本廣 夕佳, 桑形 恒男, 半場 祐子, 且原 真木, 奈良 久美

    第62回日本植物生理学会(島根)(オンライン)  2021.3 

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    Event date: 2021.3.14 - 2021.3.16

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  • PIP1型アクアポリン水輸送機能の新しい活性化メカニズム

    柴坂三根夫, 堀江智明, 且原真木

    日本植物学会第84回大会(名古屋)(オンライン)  2020.9 

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    Event date: 2020.9.19 - 2020.9.21

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  • 塩ストレス初期段階でのオオムギアクアポリンHvPIP2;1 のリン酸修飾による根水透過性 (Lpr) の下方制御機構

    大西亜耶, 且原真木

    第61回日本植物生理学会(大阪)(オンライン)  2020.3 

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  • Ca2+-sensitive and non-selective Na+/K+ channel activity in a barley aquaporin HvPIP2;8

    Sen Thi Huong Tran, Maki Katsuhara, Tomoaki Horie

    2020.3 

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    Event date: 2020.3.19 - 2020.3.21

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  • イネのTILLING 変異系統を用いたOsHKT1;4 Na+ 輸送体の塩ストレス下における生理機能の解明

    河村敏貴, 小林奈通子, グエンタンハオ, 石川亮, 且原真木, 田野井慶太朗, 松坂弘明, 熊丸敏博, センテナックヘルベ, ベリーアナリエナ, 堀江智明

    第61回日本植物生理学会(大阪)(オンライン)  2020.3 

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  • シロイヌナズナ液胞膜型アクアポリン (AtTIPs) の水輸送活性の解析

    宇都木繁子, 且原真木

    第61回日本植物生理学会(大阪)(オンライン)  2020.3 

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    Event date: 2020.3.19 - 2020.3.21

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  • Expression and Ion Transport Activity of Salt Tolerant Pokkali Rice OsHKT1;1 Variants.

    Shahin Imran, Maki Katsuhara, Tomoaki Horie

    2020.3 

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    Event date: 2020.3.19 - 2020.3.21

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  • デンジソウの就眠運動リズムにおけるアクアポリンの発現解析

    田中福人, 且原真木, 富岡憲治

    第26回日本時間生物学会学術大会(金沢)  2019.10 

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    Event date: 2019.10.12 - 2019.10.13

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  • シロイヌナズナ液胞膜型アクアポリンAtTIP2;2の過酸化水素透過性の検討

    山成由佳子, 中原由揮, 且原真木, 奈良久美

    日本植物学会第83回大会(仙台)  2019.9 

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    Event date: 2019.9.15 - 2019.9.17

    Language:Japanese   Presentation type:Poster presentation  

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  • 時計因子ELF3は胚軸における水輸送調節にどうのように関連しているか?

    藤田知美, 奥村綾子, 土平絢子, 前島正義, 且原真木, 奈良久美

    日本植物学会第83回大会(仙台)  2019.9 

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    Event date: 2019.9.15 - 2019.9.17

    Language:Japanese   Presentation type:Poster presentation  

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  • 塩生植物アッケシソウから単離した新規ペプチドの遺伝子(SeNN43)は植物の耐塩性を向上させ、根の細胞を膨張させる

    坂本光, 開沼健太, 北村蒼門, 中原由揮, 且原真木, 小栗秀

    第60回日本植物生理学会(名古屋)  2019.3 

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    Event date: 2019.3.13 - 2019.3.15

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  • 耐塩性の異なるオオムギ品種の塩ストレス初期応答

    大西亜耶, 且原真木

    第60回日本植物生理学会(名古屋)  2019.3 

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    Event date: 2019.3.13 - 2019.3.15

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  • Electrophysiological Analysis of Rice OsHKT1;1 variants

    Shahin Imran, Maki Katsuhara, Tomoaki Horie

    2019.3 

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    Event date: 2019.3.13 - 2019.3.15

    Language:English   Presentation type:Oral presentation (general)  

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  • オオムギ種子における液胞膜型アクアポリン(HvTIPs)の役割

    宇都木繁子, 且原真木

    第60回日本植物生理学会(名古屋)  2019.3 

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    Event date: 2019.3.13 - 2019.3.15

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  • オオムギの根水透過性制御にかかわる遺伝因子の探求

    大西亜耶, 且原真木

    第49回根研究集会(盛岡)  2018.10 

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    Event date: 2018.10.27 - 2018.10.28

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  • Na+/K+透過性をもつアクアポリン:イネOsPIP2;4およびオオムギHvPIP2;8の解析

    且原真木, 堀江智明

    第49回根研究集会(盛岡)  2018.10 

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    Event date: 2018.10.27 - 2018.10.28

    Language:Japanese   Presentation type:Oral presentation (general)  

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  • シロイヌナズナの液胞膜型アクアポリンAtTIP2;2とAtTIP2;3の水透過性

    山成由佳子, 中原由揮, 且原真木, 奈良久美

    日本植物学会第82回大会(広島)  2018.9 

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    Event date: 2018.9.14 - 2018.9.16

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  • シャジクモ細胞膜リン酸輸送体の分子機能解析

    三村徹郎, 大西美輪, 村西直樹, 藤原ひとみ, 石崎公康, 深城英弘, 西山智明, 坂山英俊, Rob, J.Reid, 且原真木

    日本植物学会第82回大会(広島)  2018.9 

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    Event date: 2018.9.14 - 2018.9.16

    Language:Japanese   Presentation type:Oral presentation (general)  

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  • Regulation of water transport and Na+ influx. Invited

    Katsuhara, M

    The 22nd New Phytologist Workshop ‘Energy costs of salinity tolerance in crop plants’ (Adelaide, Australia)  2018.4 

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    Event date: 2018.4.10 - 2018.4.12

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

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  • CO2輸送体候補OsTIP2;2の細胞内局在

    中原由揮, 柴坂三根夫, 谷口洋二郎, 且原真木

    第59回日本植物生理学会(札幌)  2018.3 

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    Event date: 2018.3.28 - 2018.3.30

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  • CO2輸送体候補アクアポリンOsTIP2;2の同定と機能解析

    中原由揮, 柴坂三根夫, 森泉, 谷口純一郎, 堀江智明, 金子智之, 且原真木

    第59回日本植物生理学会(札幌)  2018.3 

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    Event date: 2018.3.28 - 2018.3.30

    Language:Japanese   Presentation type:Oral presentation (general)  

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  • イネアクアポリンOsPIP2;4 の発現量と根水透過性(Lpr)の相関性

    大西 亜耶, 且原 真木

    第52回根研究集会(熊本)(オンライン)  2020.11.21 

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  • Regulation in root hydraulic conductivity (Lpr) and PIP aquaporins under salt stress.

    Katsuhara, M

    Aquaporin solutions - Innovation conference.  2020.11.10 

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    Language:English   Presentation type:Oral presentation (invited, special)  

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  • Regulation of water transport and sodium influx in plant cells Invited

    Katsuhara, M

    Jilin University College of Plant Science Symposium  2019.10.17 

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  • 塩ストレス下でのナトリウム流入と水輸送の制御 Invited

    且原真木

    第34回資源植物科学シンポジウム・第10回植物ストレス科学研究シンポジウム  2018.3.5 

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    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)  

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  • A barley HvCNGC2-3, is activated by cAMP and the co-presence of external Na+ and K+, and permeates Na+ and K+ non-selectively.

    Taiwan-Japan Plant Biology 2017  2017 

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  • ハマダイコンにおけるセシウム蓄積特性の解析

    日本植物学会第81回大会  2017 

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  • 内膜系におけるアクアポリンの輸送基質と機能

    第19回植物オルガネラワークショップ  2017 

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  • ダイコン野生種ハマダイコンにおける海岸生-琵琶湖陸封生の間での耐塩性分化

    日本植物学会第81回大会  2017 

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  • 種子におけるオオムギ液胞膜型アクアポリン(HvTIPs)の相互作用と水輸送活性

    第58回日本植物生理学会年会  2017 

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  • Identification of the CO2 transporters from among the plant aquaporins.

    Taiwan-Japan Plant Biology 2017  2017 

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  • 海浜植物ハマエンドウのエコタイプ間に見られる環境応答能力の違い

    日本植物分類学会第15回大会  2016 

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  • 地球環境問題と形質転換作物:避けて通れない課題

    倉敷地方新農業経営者クラブ連絡協議会セミナー  2016 

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  • Barley and rice aquaporins transporting hydrogenperoxide.

    第57回日本植物生理学会年会  2016 

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  • A Salicornia europaea gene (SeNN43) encodes a novel short peptide that can improve salt tolerance in plants.

    第57回日本植物生理学会年会  2016 

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  • オオムギ原形質膜型アクアポリンPIP2 ファミリーの機能と根における発現プロファイル

    日本植物学会第80回大会  2016 

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  • シロイヌナズナ概日時計変異体elf3 の根の水透過性

    日本植物学会第80回大会  2016 

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  • Preliminary dissection of CO2 permeability of a cyanobacterium.

    日本植物生理学会2014年度年会  2014 

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  • 過酸化水素輸送性アクアポリンの同定と機能/発現解析

    日本植物学会第78回大会  2014 

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  • 琵琶湖に隔離されたハマエンドウにおける耐塩性低下の検証

    植物分類学会第13回大会  2014 

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  • Plant aquaporin: A molecular regulator in plant water relations.

    The 1st Okayama University and King Faisal University's Workshop on Environmental and Life Science.  2014 

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  • 淡水域に隔離されたハマエンドウの耐塩性低下に関する比較研究

    日本植物学会第78回大会  2014 

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  • The effect of aquaporin McMIPB and RsPIP2 on leaf photosynthesis of tobacco and eucalyptus plants.

    日本植物生理学会2014年度年会  2014 

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  • 沈水植物ヤナギモにおける水輸送タンパク質PIPの働き

    日本植物生理学会2014年度年会  2014 

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  • Plasma membrane intrinsic protein1 (PIP1) is a modulator of PIP2 water channel.

    日本植物生理学会2014年度年会  2014 

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  • CO2輸送体スクリーニング法の開発

    日本植物生理学会2014年度年会  2014 

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  • 琵琶湖湖岸と海浜に生育するハマエンドウにおける環境ストレスへの適応的な応答

    日本植物生理学会2014年度年会  2014 

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  • イネOsHAK2およびOsHAK5輸送体の機能解析

    日本植物生理学会2014年度年会  2014 

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  • CO2 permeability of PIP2 aquaporins.

    日本植物生理学会2014年度年会  2014 

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  • 植物細胞の水チャネルの調節

    神谷宣郎先生生誕百周年記念シンポジウム  2013 

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  • 塩生植物アッケシソウの新規耐塩性遺伝子の機能スクリーニング

    第54日本植物生理学会年会  2013 

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  • イネの水利用、成長、アクアポリン発現におよぼす低湿度と低地温の影響

    第54日本植物生理学会年会  2013 

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  • 種子特異的に発現するオオムギTIP3;1による水輸送活性の調節

    第54日本植物生理学会年会  2013 

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  • 琵琶湖に陸封されたハマエンドウと海浜集団の間に生じた光合成特性の分化

    第54日本植物生理学会年会  2013 

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  • 琵琶湖に隔離されたハマエンドウと海浜集団の間におけるABAを介した光合成特性の分化

    日本植物学会第77回大会  2013 

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  • それではいかに放射性物質に対応するか:植物科学からの取り組み

    吉備の国クラスター・エコ環境グループ講演会  2013 

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  • 水輸送とCO2輸送の分子基盤:アクアポリン

    日本植物学会第77回大会シンポジウム  2013 

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  • 原形質膜型アクアポリンPIP1とPIP2の共発現による活性抑制

    第54日本植物生理学会年会  2013 

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  • アクアポリンのCO2透過性

    第54日本植物生理学会年会  2013 

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  • イネアクアポリンOsPIPの発現と根水透過性:浸透圧ストレスの影響および形質転換での解析

    第54日本植物生理学会年会  2013 

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  • Root hydraulic conductivity and PIP aquaporins of barely and rice in response to salinity/osmotic stress.

    植物膜生物学国際学会2013  2013 

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  • 重金属イオンによる植物アクアポリン阻害作用についての検討

    第54日本植物生理学会年会  2013 

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  • Effect of low root-temperature on water use, growth and aquaporin expression in rice plants under various air-humidity conditions.

    植物膜生物学国際学会2013  2013 

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  • The functional analysis of Cyclic-Nucleotide Gated Cation Channels (CNGCs) in barley.

    植物膜生物学国際学会2013  2013 

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  • The photosynthetic response of tobacco and eucalyptus plants overexpressing aquaporins.

    植物膜生物学国際学会2013  2013 

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  • Gating of plasma membrane intrinsic protein1 via interaction with plasma membrane intrinsic protein2.

    植物膜生物学国際学会2013  2013 

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  • The Third Type of PIP Aquaporins.

    日本植物生理学会2012年度年会  2012 

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  • オオムギとイネの根水透過性制御と浸透圧ストレス応答

    第36回根研究集会  2012 

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  • Regulation of root hydraulic conductivity in gramineous plants under osmotic stress.

    日本植物生理学会2012年度年会  2012 

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  • オオムギにおけるNa輸送性CNGCチャネルの機能解析

    日本土壌肥料学会2012年度鳥取大会  2012 

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  • Osmotic-induced reduction of root hydraulic conductivity is essential as an early response in slat/osomotic tolerance

    ゴードン科学会議 植物の塩と水ストレス  2012 

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  • イネとオオムギの亜ヒ酸輸送性NIP型アクアポリン

    日本植物学会第76回大会  2012 

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  • 水生植物ヤナギモにおける水輸送タンパク質PIPの解析

    日本植物学会第76回大会  2012 

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  • Dynamic regulation of root hydraulic conductivity and aquaporins under salt stress.

    International symposium, Strategies of Plants against Global Environmental Change.  2011 

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  • Detection of transport activity in barley and rice aquaporins with newly developed yeast system.

    日本植物生理学会2011年度年会  2011 

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  • Hydrogen peroxide permeability of aquaporin in rice, barley and Arabidopsis

    第3回植物アクアポリン研究会  2011 

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  • 形質膜アクアポリンPIP1の水チャンネルはPIP2によって開けられる

    日本植物生理学会2011年度年会  2011 

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  • アクアポリンのCO2透過性

    第3回植物アクアポリン研究会  2011 

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  • 浸透圧ストレス環境下におけるオオムギの水利用機能研究

    第3回植物アクアポリン研究会  2011 

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  • 亜ヒ酸輸送活性を持つイネおよびオオムギのアクアポリン

    第3回植物アクアポリン研究会  2011 

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  • オオムギの根水透過性とアクアポリンの浸透圧ストレス応答

    第3回植物アクアポリン研究会  2011 

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  • 原形質膜局在型アクアポリンの共発現による活性化の分子機構

    第3回植物アクアポリン研究会  2011 

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  • 浸透圧ストレス環境下におけるイネ科作物の水利用機能研究

    第35回根研究集会  2011 

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  • Dynamic regulation of root hydraulic conductivity and aquaporins under salt stress.

    The JSRR's 20th Anniversary Symposium  2011 

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  • 緑化ブロックと屋上緑化の新展開

    第二回かんきょうひろば全体交流会  2011 

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  • オオムギPIP1型と2型アクアポリンのへテロマー形成の分子機構

    日本植物生理学会2010年度年会  2010 

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  • 植物における水輸送の分子基盤・アクアポリン:動物との共通点と相違点

    3部局合同セミナー  2010 

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  • 原形質膜局在型アクアポリンPIP1とPIP2の共発現による活性化メカニズムの解析

    日本植物生理学会2010年度年会  2010 

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  • 緑化ブロックの開発

    第4回産学官民コミュニティー全国大会 in おかやま  2010 

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  • Regulation of Root Water Permeability in Barley under Salt Stress.

    ゴードン科学会議(植物の塩およびお水ストレス)  2010 

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  • Cooling effect on buildings by the roof greening at Institute of Plant Science and Resources, Okayama University.

    第5回ジョモケニアッタ農工大学科学・技術;産業会議  2010 

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  • Studies on water channel activity of a PIP1 aquaporin and its interaction with PIP2 aquaporins.

    第15回国際植物膜生物学ワークショップ  2010 

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  • Water transport in plants under stresses: from molecules to whole plant.

    第5回ジョモケニアッタ農工大学科学・技術;産業会議  2010 

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  • 高浸透圧ストレス下におけるオオムギ根の水輸送活性制御機構の解明

    日本植物生理学会2010年度年会  2010 

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  • Aquaporin - molecular mechanism of water transport in plants: Common and different characteristics between animals and plants.

    2010 

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  • 水と植物の2つの視点から 見えること 考えること

    第四回岡山大学いちょう並木研究サロン  2009 

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  • 原形質膜局在型アクアポリンPIP1とPIP2の共発現による活性化メカニズムの解析

    日本植物生理学会2009年度年会  2009 

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  • オオムギアクアポリンの機能と塩ストレス応答

    日本植物生理学会2009年度年会  2009 

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  • オオムギ根の水輸送活性は塩ストレス下において翻訳後修飾によって抑制されている

    日本植物生理学会2009年度年会  2009 

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  • Varietal differences in the rates of root growth and leaf photosynthesis, hydraulic conductivity and expressions of aquaporins in barley seedlings under salt stress conditions.

    PlantBiology2009  2009 

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  • 花と果実におけるアクアポリンの発現・調節と機能

    日本植物生理学会2009年度年会  2009 

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  • アクアポリンが光合成機能に果たす役割の定量的評価

    日本生態学会近畿地区会2009年第2回例会  2009 

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  • 高等植物の光合成と水チャネル

    第55回日本生態学会福岡大会  2008 

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  • マメゾウムシ二種におけるオスによるメスの交尾抑制の変異と種間交尾がメスの再交尾に与える影響

    第55回日本生態学会福岡大会  2008 

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  • Xenopus oocyteとPichia pastorisで発現させたチューリップ花弁由来アクアポリンの水輸送活性:ホモログTgPIP2;2はリン酸化によって制御される

    日本植物生理学会2008年度年会  2008 

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  • Water transport in rice and Graminean plants: from whole plant to aquaporins.

    Beijing Botanical Society Workshop  2008 

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  • 草本植物の茎葉では機械的ストレスを水分移動によって緩和している

    日本植物生理学会2008年度年会  2008 

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  • オオムギ原形質膜型アクアポリンの発現制御と水輸送特性

    日本植物生理学会2008年度年会  2008 

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  • マメゾウムシ2種におけるオスの交尾抑制物質の生化学的分析

    第52回日本応用動物昆虫学会大会  2008 

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  • セイヨウナシアクアポリンPcPIP2;2のリン酸化による水輸送活性調節

    日本植物生理学会2008年度年会  2008 

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  • タバコアクアポリンの機能・発現と根の水透過性の制御

    第28回根研究会  2008 

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  • 屋上緑化の現状と展望:資源生物科学研究所の取り組みと研究開発

    吉備の国クラスター・エコ環境グループ講演会  2008 

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  • 植物と水と私たち

    第53回大原孫三郎總一郎記念講演会  2008 

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  • オオムギ根における水透過性測定のためのpressure chamber作成と測定法の確立

    第28回根研究会  2008 

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  • プロジェクト概要と現在の活動について

    岡山大学資源生物科学研究所屋上緑化プロジェクト公開報告・見学会  2008 

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  • コンクリート製緑化ブロック

    平成20年度第1回TLOシーズ説明会  2008 

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  • Transcriptional Regulations and Water Transport Activity of PIP-type Aquaporins in Roots of Barley.

    Gordon Research Conference - Salt & Water Stress In Plants  2008 

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  • Phosphorylational regulation of plasma membrane aquaporin, PIP2, and its physiological roles in fruit and flower.

    FESPB2008  2008 

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  • 塩ストレス条件下におけるオオムギ幼植物の根の水伝道度と光合成速度の品種間差

    日本作物学会第226回講演会  2008 

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  • PIP aquaporins and water relations in barley roots under salt stress.

    Gordon Research Conference ? Salt & Water Stress In Plants  2008 

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  • セイヨウナシのアクアポリンPIP2のリン酸化による活性調節と果実日肥大におけるリン酸化状態の変化

    園芸学会平成20年度秋季大会  2008 

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  • オオムギ原形質膜型アクアポリンの発現と機能および塩ストレスと重金属の影響の解析

    日本植物学会第72回大会  2008 

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  • 塩ストレス条件下におけるオオムギ根の水透過性

    第29回根研究会  2008 

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  • 耐塩性の異なるオオムギ2品種の根における原形質膜型アクアポリンの発現

    第29回根研究会  2008 

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  • シロイヌナズナの浸透圧ストレス誘導性糖トランスポーターERD6の機能解析

    第31回日本分子生物学会年会・第81回日本生化学会大会 合同大会  2008 

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  • オオムギ液胞膜型アクアポリン遺伝子の環境ストレスによる発現調節

    日本植物生理学会2007年度年会(愛媛) 2007年3月28日-30日  2007 

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  • Aquaporins mediate the transports of essential molecules in plants growing in various environments.

    5th International Conference of Aquaporin  2007 

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  • The effect of aquaporin HvPIP2;1 and McMIPA on leaf photosynthesis for rice and tobacco plants.

    5th International Conference of Aquaporin (Nara, Japan) 2007年7月13日-16日  2007 

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  • オオムギ原形質膜型アクアポリン遺伝子の同定と塩ストレスによる発現制御

    日本植物学会第71回大会  2007 

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  • Multi-functional plant aquaporins mediating transports of water and essential molecules for plant life.

    Special Lectures by Professors in Sister Research (Agricultural Plant Stress Research Center, Chonnam National University, Korea)  2007 

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  • 緑化ブロックと建物緑化新技術の開発

    岡山大学知恵の見本市2007  2007 

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  • オオムギ原形質膜型アクアポリン遺伝子の同定と塩ストレスによる発現制御

    第27回根研究会  2007 

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  • アズキゾウムシにおけるオス由来のメス交尾抑制物質の生化学的分析

    第51回日本応用動物昆虫学会  2007 

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  • 植物のアクアポリンの多彩な役割とその制御

    第9回植物生体膜シンポジウム「植物の生理機能と生理学的構造の役割」(  2007 

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  • イネ科NIPのケイ酸輸送特性の解析

    日本植物生理学会2007年度年会  2007 

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  • オオムギのアルミニウム活性型クエン酸輸送体候補遺伝子の発現ならびに機能解析

    日本植物生理学会2007年度年会  2007 

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  • 塩ストレスの初期段階におけるイネアクアポリン発現の同調的変動

    日本植物生理学会2007年度年会(愛媛) 2007年3月28日-30日  2007 

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  • 在来および帰化マンネングサ属の水利用効率の比較

    日本植物生理学会2007年度年会  2007 

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  • Cloning and functional analysis of an aluminium-induced malate transporter gene from rape (Brassica napus).

    日本植物生理学会2006年度年会  2006 

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  • 水輸送の分子機構・アクアポリンと低温

    第52回低温生物工学会セミナー  2006 

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  • 水環境とオオムギの遺伝子発現制御

    日本植物生理学会2006年度年会  2006 

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  • A molecular investigation of an aluminium-induced malate secretion from rape (Brassica napus L.).

    PlantBiology2006  2006 

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  • Salt and osmotic stress regulates differentially plasma-membrane-type aquaporin gene expression in salt tolerant and salt sensitive barley.

    Gordon Research Conference – Salt & Water Stress In Plants  2006 

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  • Cloning and Functional Analysis of Plasma Membrane Aquaporin Subfamilies of Tulip Petal.

    PlantBiology2006  2006 

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  • ヒ素と重金属に応答する植物モニター系の構築

    岡山大学・学内COE「資源生物を用いた地球環境のモニター系の構築と環境保全への応用」国際ワークショップ  2006 

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  • アクアポリンの水ストレスへの応答と多彩な機能

    東北農業研究セミナー「生命を支える分子システム-植物の水吸収をになう「アクアポリン」研究の最前線」  2006 

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  • アクアポリン分子の基質輸送に伴う分子骨格構造の変動

    日本植物生理学会2006年度年会  2006 

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  • アクアポリンの多様性と多面性:耐塩性や光合成との関係 (シンポジウム アクアポリンの多様性:構造、機能、局在)

    日本植物生理学会2005年度年会シンポジウム  2005 

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  • イネゲノム中の全てのPIPファミリー遺伝子

    日本植物生理学会2005年度年会  2005 

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  • Transcriptional regulation of plasma membrane intrinsic protein (PIP) aquaporins in barley (Hordeum vulgare ,Haruna-nijo) by heavy metals.

    10th International Congress SABRAO  2005 

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  • Functional and molecular analysis of the barley Ca2+/H+ antiporter.

    Plant Biology2005  2005 

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  • オオムギの液胞膜型Cation/H+ exchanger (CAX)

    第8回植物生体膜シンポジウム  2005 

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  • 岡山県南部に適化した薄層省管理緑化の新システムの開発

    大学発新事業創出促進事業「技術シーズ発信会」  2005 

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  • オオムギのCDPK遺伝子および液胞膜型CAX遺伝子の同定と機能解析

    第69回日本植物学会  2005 

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  • Osmotic stress regulates differentially aquaporin genes expression in salt tolerant and salt sensitive barley (Hordeum vulgare).

    4th Plant Genomic European Meetings  2005 

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  • 水チャネル・MIP遺伝子ファミリーの多様な機能

    第1回ムギ類研究会  2005 

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  • オオムギ根のアクアポリンと水輸送、および塩ストレスとの関係

    第68回日本植物学会  2004 

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  • Aquaporins under salt stress

    Gordon Research Conference "Salt and Water Stress in Plants"  2004 

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  • オオムギにおけるストレス応答性遺伝子MAPKおよびCAXの単離と機能解析

    第21回根研究集会  2004 

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  • グルタチオン包合体の液胞膜透過機構に関するABCトランスポーター遺伝子

    第68回日本植物学会  2004 

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  • 植物のアクアポリンの働き

    第19回「大学と科学」公開シンポジウム みずみずしい体のしくみ:水の通り道“アクアポリン”の働きと病気  2004 

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  • オオムギ根の水輸送とアクアポリンファミリー

    第21回根研究集会  2004 

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  • 植物細胞におけるイオンと水:その生理と輸送機構の動物細胞との共通点と相違点

    第47回日本腎臓学会学術総会  2004 

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  • 屈性を示している根の細胞の水吸収

    第1回植物アクアポリン研究討論会  2004 

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  • オオムギ水チャネルとその塩ストレス応答についての解析

    第1回植物アクアポリン研究討論会  2004 

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  • オオムギアクアポリンHvPIP2;1を過剰発現させた形質転換イネ葉での二酸化炭素拡散コンダクタンスと光合成速度の上昇

    第1回植物アクアポリン研究討論会  2004 

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  • Regulation of malate efflux in wheat root apex under aluminum stress: role of protein phosphorylation and ALMT1 gene.

    The 1st International Symposium "Life and Environmental Science in Future"  2004 

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  • 水チャンネル過剰発現イネにおける根の形質と塩ストレス応答性の変化

    日本植物生理学会2004年度年会  2004 

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  • Increase in leaf CO2 conductance and decrease in salt tolerance in transgenic rice plant over-expressing barley aquaporin.

    Plant Biology 2003  2003 

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  • タバコ由来のグルタチオン-S-トランスフェラーゼ遺伝子(parB)の導入によって活性酸素除去系を強化した形質転換シロイヌナズの塩ストレスに対する応答

    日本植物学会第67回大会  2003 

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  • オオムギ幼根の水透過性に関連する水チャネルの夜間における発現

    第19回根研究集会  2003 

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  • オオムギ水チャンネルは二酸化炭素を透過する-形質転換イネ葉での二酸化炭素拡散コンダクタンスの上昇-

    関西光合成研究会  2003 

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  • オオムギの水輸送と塩ストレス応答

    第19回根研究集会  2003 

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  • 重金属イオン輸送体、CPx-ATPase遺伝子(Bxa1)の単離と特性解析

    日本植物生理学会2003年度年会  2003 

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  • コムギにおけるアルミニウム活性化型リンゴ酸トランスポーター遺伝子のアルミニウム耐性への関与

    日本植物生理学会2003年度年会  2003 

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  • コムギのAl耐性機構-リンゴ酸分泌とそのトランスポーター遺伝子-

    第18回根研究集会  2003 

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  • オオムギ水チャンネル遺伝子を過剰発現させた形質転換体イネ緑葉での二酸化炭素透過性(拡散コンダクタンス)の上昇

    日本植物生理学会2003年度年会  2003 

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  • コムギのアルミニウム活性化型リンゴ酸トランスポーター遺伝子の機能解析:対立遺伝子間の機能相違

    日本土壌肥料学会2003年度神奈川大会  2003 

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  • かび臭物質生産ラン藻Oscillatoria brevisの重金属耐性に関する分子生物学的研究

    日本水処理生物学会第39回大会  2002 

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  • アルミニウム耐性小麦におけるリンゴ酸放出機構の分子生物学的解析

    第6回植物生体膜シンポジウム  2002 

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  • 塩ストレス抵抗性オオムギに関する研究1. 塩ストレスで特異的に発現する遺伝子の解析

    日本農芸化学会2002年度大会  2002 

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  • 植物水チャンネルの遺伝子と機能

    第6回植物生体膜シンポジウム  2002 

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  • アルミニウム耐性小麦に特異的な遺伝子の機能解析

    日本土壌肥料学会2002年度名古屋大会  2002 

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  • オオムギの水チャネルHvPIP2;1の塩ストレスおよび乾燥ストレス下における機能的発現と日周変化

    日本植物生理学会2002年度年会  2002 

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  • かび臭物質生産ラン藻Oscillatoria brevisの多種金属耐性に関わる重金属イオン輸送体に関する研究

    日本水処理生物学会第39回大会  2002 

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  • ラン藻Oscillatoria brevisのメタロチオネイン遺伝子の同定

    日本植物学会第65回大会  2001 

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  • ラン藻Oscillatoria brevisからの重金属輸送遺伝子の同定と解析

    日本植物生理学会2001年度年会  2001 

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  • オオムギ根から単離した水チャンネル遺伝子の機能発現

    日本植物生理学会2001年度年会  2001 

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  • ラン藻Oscillatoria brevisのP-型重金属イオン輸送ATPaseの研究

    日本植物学会第65回大会  2001 

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  • オオムギの水チャンネル遺伝子bpw1の局在性と発現の日周変化

    日本植物生理学会2001年度年会  2001 

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  • Water channels in barley roots.

    The 6th International Society of Root Research Symposium  2001 

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  • 塩ストレス誘導性細胞死と生体膜

    植物生体膜シンポジウム  2000 

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  • 植物の水チャンネルの構造と機能発現

    第17回資源生物科学シンポジウム  2000 

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  • Apoptosis-like cell death under salt stress.

    Gordon Research Conference "Cellular Basisi of Adaptation to Salt and Water Stress in Plants"  2000 

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  • Cell death and stress response in plants

    International symposium on "Stress responses in biological systems"  1999 

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  • 塩ストレス誘導性細胞死の初期過程における細胞膜インテグリティーの研究

    日本植物生理学会1999年度年会  1999 

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  • オオムギからの水チャンネル遺伝子の単離と塩ストレス下における発現の解析

    日本植物生理学会1999年度年会 1999年3月  1999 

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  • 耐塩性の異なるオオムギ3品種におけるsalT様遺伝子の発現制御

    日本植物学会第63回大会  1999 

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Works

  • 都市緑化おかやまフェア2009

    2009

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Awards

  • 第5回村川技術奨励賞

    2010  

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    Country:Japan

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  • 日本作物学会論文賞

    2007  

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    Country:Japan

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  • 日本植物生理学会論文賞

    2006  

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    Country:Japan

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Research Projects

  • Remediation of the agricultural environments using the plant species with roots containing high H2O2 in the presence of woody plant organics-Fe complex via rhizosphere Fenton reaction

    Grant number:21K05869  2021.04 - 2024.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    我妻 忠雄, 且原 真木, 俵谷 圭太郎, 田原 恒

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    Grant amount:\4160000 ( Direct expense: \3200000 、 Indirect expense:\960000 )

    キク科植物根のみが過酸化水素(H2O2)を大量に放出し、周辺鉄(Fe)との根圏Fenton反応でメタン、アトラジンを分解できることを見出した(Wagatsuma et al. 2021, 2022)。以上を踏まえ実験し、以下の結果を得た:(1)キュウリ、オオムギ、マリーゴールド(キク科)幼植物根を、H2DCF(細胞内活性酸素に蛍光)とBESH2O2(細胞内H2O2に蛍光)で、根アポプラスト中のH2O2をKI + starch溶液で、それぞれ染色した。その結果、オオムギはH2O2が不検出、キュウリ、マリーゴールドは細胞内に高濃度のH2O2を持つが、マリーゴールドのみが根外にH2O2を放出することが解った。(2)高塩環境で進化発生したキク科植物根から大量に放出されるH2O2は、根圏土壌catalaseでO2とH2Oに速やかに分解され、塩類土壌根圏に不足するO2の補給、塩類濃度の希釈、土壌緻密度の降下などによって塩ストレス回避に貢献すると推察した。(3)間伐材phenolicsからのFenton反応用の高性能鉄資材の創製のために、まずユーカリ葉を用いた。70 % 含水アセトン抽出液中の水溶液にFeCl3を反応させ、Phenolics-Fe錯体を得た。この凍結乾燥粉末の水溶性画分には、有機物Homologueとして一般に用いられているメチレンブルー(MB)をH2O2存在下で分解するFenton活性が、不溶性画分にはFenton活性とMB吸着能が認められ、これら能力には反復性があった。今後の方向性は、以下の通りである:①より高濃度のH2O2を放出するキク科植物を探索する。②H2O2供給土壌のcatalase活性の変化を明らかにすること。③今回創製したPhenolics-Fe錯体よりも、より強力な活性を持つ鉄資材の創製に取り組むこと。④土壌腐植、各種有機質肥料・資材などに対するFenton反応による小分子化の実態を明らかにすること。⑤以上により、Fenton活性による根圏有機環境の制御によって、有害有機環境を修復するとともに、高塩や低リンなどのストレスを回避させ得るかを解明すること。

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  • デュアル輸送機能を示す新型アクアポリンの分子機構と生理機能の解明

    Grant number:20K06708  2020.04 - 2024.03

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    且原 真木, 堀江 智明

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    Grant amount:\4420000 ( Direct expense: \3400000 、 Indirect expense:\1020000 )

    本研究では水輸送機能と同時にナトリウム輸送機能も合わせ持つ、すなわち2つの異なる輸送機能をデュアルで示す新型アクアポリンを対象として、分子生理学および電気生理学的手法によって機能特性を解明すること、また免疫組織化学や形質転換体を用いて生理機能を明らかにすることを目指している。
    本年度はイネの原形質膜型アクアポリンPIP11種をアフリカツメガエル卵母細胞に発現させて二電極電圧クランプ法を適用して解析した。その結果イネ原形質膜型アクアポリンの一つOsPIP2;4がK+やNa+を透過させることを見出した。OsPIP2;4は葉より根での発現が高く、間接蛍光抗体法により主として外皮で発現していることがわかった。OsPIP2;4は、細胞外Ca2+濃度が低いときは一価カチオンに対する輸送活性を示した(透過性の順序はK+ > Na+ > Rb+ ~ Cs+ > Li+)が、高Ca2+条件ではイオン輸送活性は阻害された(IC50=92μM)。OsPIP1型アクアポリンと共発現させると、OsPIP2;4のイオン輸送性は阻害された。人為的にアミノ酸を置換した変異OsPIP2;4の解析から、水輸送とイオン輸送は分子内で別々の機構によって制御されていると考えられた。すなわち、水を輸送するが一価カチオンを輸送しない変異OsPIP2;4分子や、逆に水を輸送できないが一価カチオンを輸送する変異OsPIP2;4分子を得ることができた。またOsPIP2;4による過剰発現体と抑制体のイネ植物体を使って塩ストレス下でのNaやKの蓄積量を調べた結果、野生型イネと比べて過剰発現体では150 mM NaClの塩ストレスによってNaの蓄積が増加する一方で抑制体ではNaの蓄積が減少することが明らかになった。

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  • A Novel Therapeutic Strategy for Macular Edema Targeting Aquaporins.

    Grant number:17K11450  2017.04 - 2020.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    Shiraga Fumio

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    Grant amount:\4680000 ( Direct expense: \3600000 、 Indirect expense:\1080000 )

    Macular edema is a common condition in diabetic retinopathy, retinal vein occlusion, age-related macular degeneration, and macular epiretinal membranes. Excessive changes in water and osmolality impair the cells that make up the macula, resulting in decreased vision. In this study, we focused on retinal pigment epithelial cells, which are responsible for water transport in the macula, and aquaporins, which are water transport proteins in cells, and investigated the effects of various stresses on cells in macular diseases on the expression of aquaporins in retinal pigment epithelial cells. The results showed that the expression of aquaporin-1 was significantly decreased by extension stress on the cells. On the other hand, there were no significant changes in the expression of other aquaporins.

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  • Physiological and molecular study of CO2 permeable aquaporins

    Grant number:26440146  2014.04 - 2017.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    Katsuhara Maki, MORI Izumni

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    Grant amount:\5200000 ( Direct expense: \4000000 、 Indirect expense:\1200000 )

    We established yeast screening system to identify CO2 permeable aquaporins. Among 69 aquaporins from rice, barley and Arabidopsis, 18 aquaporins were identified as putative CO2 permeable aquaporins. One of these aquaporins, OsTIP2;2 was detected around chloroplasts and some organelles (possible mitochondria), and seemed to have a function to supply CO2 intracellularly to inner chloroplast.

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  • Identification and analysis of rice and barley aquaporins as arsenite transporters.

    Grant number:20580063  2008 - 2010

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    KATSUHARA Maki

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    Grant amount:\4420000 ( Direct expense: \3400000 、 Indirect expense:\1020000 )

    In this project, rice and barley aquaporins were selected and analyzed form the view to elucidate the molecular mechanism of arsenite transports in Poaceae plants. Final goal of this project is to breed rice/barley lines containing low As. Using screening system with yeast cells, 5 aquaporin genes (including known 3 genes) were identified as arsenite transporters. Among these genes, temporal increase of OsNIP2 ; 1 expression in roots and decrease of OsNIP3;3 in leaves were observed in rice plants treated with arsenite.

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  • Studies on Functional Analysis and Regulation Mechanism of Plant Aquaporins

    Grant number:19580106  2007 - 2008

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    SHIBATA Hitoshi, KATSUHARA Maki

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    Grant amount:\3250000 ( Direct expense: \2500000 、 Indirect expense:\750000 )

    チューリップ花弁の開閉は温度依存的であり、開花のためには水の花弁への輸送が必須で、水チャンネルの活性化が起こる。水チャネルを構成するアクアポリンの分子種を同定するために、細胞質アクアポリン遺伝子4種類をクローニングし、TgPIP2:2が水の輸送に関与することを機能面から確証した。またアクアポリンがリン酸化されて活性型になり、脱リン酸化で不活性化されることを確認した。TgPIP2:2も分子内でリン酸化を受ける3種のセリン残基(Ser35, Ser116, Ser274)を特定した。また、PIP2:2が花弁、茎、葉、塊茎、根いずれの組織においても最も発現量が多い分子種であり、根から花弁にいたる水の輸送に関与している可能性を指摘した。
    TgPIP2-2を酵母(ピキア)に発現させ、水チャンネル活性を図る手法を新たに導入した。リン酸化脱リン酸化の阻害剤が活性に影響したことから、簡便に水チャンネル活性を測定できる系であるとともに調節機構を検証するための優れた方法であると結論した。

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  • The elucidation of the low-temperature stress mechanism using reconstituted vesicles with tonoplast H^+-ATPase

    Grant number:11440237  1999 - 2001

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    KASAMO Kunihiro, KATSUHARA Maki, SHIBASAKA Mineo

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    Grant amount:\14200000 ( Direct expense: \14200000 )

    Proton pumping across the tonoplast was markedly suppressed by chilling. In order to clarify the mechanism of chilling-induced depression of proton-pumping across tonoplast. we reconstituted chimera proteoliposomes with H^+-ATPase solubilized from the tonoplast of chilling insensitive rice (var. Nipponbare) without chilling and individual or combined lipids extracted from the tonoplast of chilling-sensitive rice (var. Boro) cells with or without chilling at 5 ℃. We further examined the relation between membrane fluidity and chilling.
    As the result, the chilling-induced decrease in proton pumping across the tonoplast is due to the decrease in the fluidity of the hydrophilic surface region of the lipid bilayer of the tonoplast, which is caused by glycolipids, such as glucocerebroside and acylsteryl glucoside.

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  • 塩ストレスによる細胞死誘導機構

    Grant number:10740370  1998 - 1999

    日本学術振興会  科学研究費助成事業  奨励研究(A)

    且原 真木

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    Grant amount:\2200000 ( Direct expense: \2200000 )

    プログラム細胞死(アポトーシス)は最近の研究から、植物細胞において道管形成などの形態形成や病原菌への抵抗性などで、中心的な役割をになう重要な細胞機構であると考えられるようになってきた。しかしその初期過程および分子メカニズムについてはまだ不明の点が多い。そこでこれらの点について、塩ストレスによって細胞死が誘導されるオオムギ根の実験系で研究をおこなった。
    動物細胞のアポトーシスにおいては、核DNAの断片化に先立って細胞膜インテグリティーの崩壊が起こるとされている。すなわち、通常では細胞膜脂質二重層の細胞質側に局在するフォスファチジルセリン(PS)の局在がくずれ、細胞外からPSに特異的に結合するアネキシンVによって細胞が標識されるようになる。昨年度の研究ではオオムギ根、オオムギ培養細胞、およびイネ培養細胞から単離したプロトプロストでは、塩ストレス処理によってはアネキシンVで標識されるようにはならない、という結果を報告した。今年度はプロトプロスト化しない、組織のままの根端を用いて、染色液を加圧して処理することにより染色することができることを見出した。すなわち塩ストレス処理後30分〜1時間で、一部の細胞がアネキシンVで標識されるようになることを明らかにした。
    また細胞死抑制遺伝子として、いくつかの植物種からdad1遺伝子が報告されたので、オオムギにおいても、そのホモログを検索した。PCR法を用いていくつかの産物を検出したが、塩基配列を解析した結果、既存のdad1と類似するものは見つからなかった。引き続きオオムギdad1の同定を進めているところである。

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  • 塩ストレスによる細胞死とストレスに適応した新根形成機構に関する細胞生理学的研究

    Grant number:09251213  1997

    日本学術振興会  科学研究費助成事業  重点領域研究

    且原 真木, 柴坂 三根夫

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    Grant amount:\1800000 ( Direct expense: \1800000 )

    塩ストレスによってオオムギ根の伸長が抑制された時、塩ストレスの程度が中程度であったり、一過的であった場合は、新根が発生する。一方、この時いったんストレスを受けて伸長が停止した古い根については、根細胞では核DNA分解を伴う細胞死が起こっていることをこれまで明らかにした。この細胞核DNAの分解は、アポトーシス様の機構が働き、時間的にも早い反応であることから、何か積極的な細胞生理的意味があるのではないかと考えた。具体的には、ストレスを受けた根の物質を分解し、輸送して塩ストレスに適応した新しい根や、地上部の形成に再利用しているのではないかと仮説を立てた。この仮説を検証するために、実験をおこなった。また細胞死と根形成の関係についてさらなる知見を得るため、ヒトおよび線虫の細胞死抑制遺伝子を導入した形質転換タバコを用いて実験をおこなった。
    オオムギ根に^<32>Pまたは^3Hで標識したチミジンを根に取り込ませて、根細胞のDNAを標識した。一定の時間根に塩ストレスを与えた後、アポトーシス様の機構で分解された核酸成分が転流・再利用されて、新根あるいは地上部の核酸合成に利用されるまで待った後、新根、地上部から核酸を抽出して、取り込まれた放射能を測定した。その結果、ストレス解除後成長を再開した地上部に有意に標識が入ることが明らかになった。新根にも標識が入るようだが、地上部ほど明確な結果にならなかった。
    次にヒトおよび線虫の細胞死抑制遺伝子を導入した形質転換タバコを農水省農業生物資源研究所から提供を受けて、根の形態形成を観察した。形質転換タバコの根は成長が遅くなり、外見上はまっすぐに伸びず、縮れたようになっていた。維管束系を観察したところ、木部の構造に異常が生じた根が多く見られ、道管の通導が悪く、成熟が遅延していることがわかった。細胞死抑制因子が働いて、木部の組織分化・細胞配列に影響を与えた結果と推察された。これらの個体の塩ストレス応答について、現在研究を進めているところである。

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  • 高親和性カリウム輸送系の細胞生理学的研究

    Grant number:08740620  1996

    日本学術振興会  科学研究費助成事業  奨励研究(A)

    且原 真木

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    Grant amount:\900000 ( Direct expense: \900000 )

    本研究では資源作物オオムギからの高親和性カリウム輸送系遺伝子の同定し,環境および植物体の組織による高親和性カリウム輸送系発現の調節を解明することを目指した。
    酵母から数種、植物ではコムギから1種類報告されていた高親和性カウム輸送系遺伝子において認められた共通のアミノ酸配列部分に対応する20から30の塩基配列を人工合成した。上流プライマーとして5種類(TRK1、TRK3、TRK5、MH1、MH2)作成した。下流プライマーとして4種類(TRK2、TRK4、HR2、HR3)作成した。
    高親和性カリウム輸送系は、カリウム欠乏条件で発現していると考えられている。そこで標準の培養液からカリウム塩を除いた条件で、2週間水耕栽培したオオムギ(品種赤神力)の根からRNAを抽出し、cDNAに変換した後、上記プライマーを上流、下流いろいろな組み合わせを使って高親和性カリウム輸送系遺伝子のPCRによる増幅を試みた。様々なPCR条件を検討したが結局いかなる組み合わせを用いてもPCR産物を得ることができなかった。名古屋大学のグループによると(未発表データ)、アラビドプシスにおいて、TRK1(5'-GGIAAYACIYTITTYCC-3')とTRK2(5'-VDIGGIARRTACATCAT-3')の組み合わせで、高親和性カリウム輸送系遺伝子を増幅することができたという。オオムギでは、このプライマーの組み合わせでもPCR産物は得ることができなかったので、アラビドプシスとは、遺伝子の塩基配列が多少異なるのかも知れない。あるいは、今回用いた栽培条件では高親和性カリウム輸送系が発現していなかったという可能性もあるので、現在栽培条件を変えて(0.25mMの硫酸カルシウムのみの培地)1週間育てたオオムギからRNAを抽出して、再度PCRをおこなっている。

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  • カリウム輸送系の細胞生理学および分子生物学的研究

    Grant number:07740622  1995

    日本学術振興会  科学研究費助成事業  奨励研究(A)

    且原 真木

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    Grant amount:\900000 ( Direct expense: \900000 )

    カリウムは植物の3大栄養の一つであり、また高濃度NaClが存在する塩ストレス条件下ではナトリウムイオンと拮抗するカリウムイオン吸収能力が耐塩性機構に深く関与すると考えられている。そこでカリウム輸送系の細胞生理学および分子生物学的研究を、Mesembryanthemun crystallinumおよびオオムギを研究材料としておこなった。前者については、カリウムチャンネル遺伝子断片がPCR法によって同定されていたので、その成果に基づき今年度は遺伝子の全長と遺伝子発現について調べた。その結果この遺伝子のcDNAは2,800bpの長さであることがわかった。またこの遺伝子は主として根で発現していた。PCR法によると、葉では根のものと異なった長さのDNAが増幅されてきたので、根とは異なる種類のカリウムチャンネルが発現しているものと考えられる。塩ストレスによる影響については、この遺伝子の発現は塩ストレスによりやや減少することが明らかになった。発現部位と塩ストレスの影響についてさらに詳細に検討するため、in situ PCR法の導入を現在検討している。つぎにオオムギのカリウム輸送系遺伝子については、Mesembryanthemun crystallinumと同じ条件では、目的とする遺伝子はPCRによって増幅できなかった。そこでPCRの条件を検討した結果、いくつかのPCR産物を得ることができた。これらが目的とするものかどうか、現在クローニングと塩基配列決定の準備を進めている。

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  • Studies on Salinity Tolerance of mineral translocation in halophyte roots.

    Grant number:06660080  1994 - 1995

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for General Scientific Research (C)

    SHIBASAKA Mineo, KATSUHARA Maki, NAKASHIMA Susumu

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    Grant amount:\1900000 ( Direct expense: \1900000 )

    This research project was aimed on determining the characteristics of plasmamembrane contributed to salinity tolerance of a halophyte, Salicornia virginica, roots, based on a physiological and biochemical comparison with mesophyte. Though integrity of mesophyte plasmamembranes has been known to be loss under conditions of high concentration of salt, it has not been clear that what changes in the membranes causes the damage. We planed to investigate what changes in the membranes enhanced permeability and how much important metabolites flowed through the membranes.
    Treatments of high NaCl conc.solutions diminished absorption of potassium of a mesophyte, Hordeum vulgare. By the contrast, these treatments enhanced that of a halophyte, Salicornia. We found that important metabolites flowed out of barley roots treated by high NaCl conc.solutions and the high energy bonds was lost. Decrement of potassium uptake in barley could be caused by the loss of energy, which was determined by ^<31>P-NMR and a biochemical quantitation. The loss of membrane integrity was detected by taking up a fluorescent substance (propidium iodide). Releasing peripheral proteins from the membranes and decreasing of plasmamembrane ATPase apoprotein were found simultaneously. However, relationship between these phenomenon and increment of membrane permeability has not be clarified.
    Because gene expression needs energy, introducing salinity tolerant gene to barley will be a failure without improving plasmamembrane.

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