2024/03/28 更新

写真a

スエヨシ クニ
末吉 邦
SUEYOSHI Kuni
所属
理事室 理事
自然科学研究科 生命・食料科学専攻 教授
農学部 農学科 教授
職名
理事
外部リンク

学位

  • 学術博士 ( 1989年3月   千葉大学 )

研究分野

  • ライフサイエンス / 植物栄養学、土壌学

経歴(researchmap)

  • 新潟大学   農学部   教授

    2012年4月 - 現在

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  • 新潟大学   自然科学研究科 生命・食料科学専攻   教授

    2012年4月 - 現在

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  • 新潟大学   農学部 応用生物化学科   准教授

    2007年4月 - 2012年3月

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  • 新潟大学   農学部 応用生物化学科   助教授

    1998年3月 - 2007年3月

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  • ワシントン州立大学   客員研究員

    1992年6月 - 1994年3月

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    国名:アメリカ合衆国

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  • 神戸大学農学部   助手

    1989年4月 - 1998年2月

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▶ 全件表示

所属学協会

留学歴

  • ワシントン州立大学   客員研究員

    1992年6月 - 1994年3月

 

論文

  • Effects of Application of Various forms of Nitrogen on the Growth of Soybean Nodules and Roots Related to the Carbon and Nitrogen Metabolism

    Takuji Ohyama, Sayuri Tanabata, Norikuni Ohtake, Takashi Sato, Kuni Sueyoshi, Yoshihiko Takahashi, Shinji Ishikawa, Yuki Ono, Natsumi Yamashita, Akinori Saito

    Soybean - Recent Advances in Research and Applications   2022年9月

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    掲載種別:論文集(書籍)内論文   出版者・発行元:IntechOpen  

    Soybean plants require a large amount of nitrogen either from nitrogen fixation in nodules or nitrogen absorption from roots. It is known that nitrate, a major inorganic nitrogen compound in upland soils, represses nodule growth and nitrogen fixation. Rapid and reversible inhibition of nodule growth and nitrogen fixation activity was found in the hydroponically cultivated soybeans after changing the nutrient solution with or without nitrate. Isotope tracer analysis revealed that the major cause of this inhibition depended on the changes in the partitioning of photo-assimilate between nodules and roots and was not directly related to the transported N compounds. Transcriptome and metabolome analyses supported that nitrate strongly promotes nitrogen and carbon metabolism in the roots but represses them in the nodules. The application of ammonium, glutamine, or urea also inhibited the nodule growth and nitrogen fixation like nitrate, although the inhibition was lower than that of nitrate. The degree of inhibition was related to the decrease in carbon isotope partitioning into the nodules, rather than the import of nitrogen isotope to nodules. Urea was detected in xylem sap and all parts of soybean, and some urea might be originated from ureide degradation.

    DOI: 10.5772/intechopen.105348

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  • Rice amino acid transporter-like 6 (OsATL6) is involved in amino acid homeostasis by modulating the vacuolar storage of glutamine in roots. 国際誌

    Saori Ogasawara, Masataka Ezaki, Ryusuke Ishida, Kuni Sueyoshi, Shunya Saito, Yuki Hiradate, Toru Kudo, Mitsuhiro Obara, Soichi Kojima, Nobuyuki Uozumi, Kentaro Tanemura, Toshihiko Hayakawa

    The Plant journal : for cell and molecular biology   107 ( 6 )   1616 - 1630   2021年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Glutamine is a product of ammonium (NH4 + ) assimilation catalyzed by glutamine synthetase (GS) and glutamate synthase (GOGAT). The growth of NH4 + -preferring paddy rice (Oryza sativa L.) depends on root NH4 + assimilation and the subsequent root-to-shoot allocation of glutamine; however, little is known about the mechanism of glutamine storage in roots. Here, using transcriptome and reverse genetics analyses, we show that the rice amino acid transporter-like 6 (OsATL6) protein exports glutamine to the root vacuoles under NH4 + -replete conditions. OsATL6 was expressed, along with OsGS1;2 and OsNADH-GOGAT1, in wild-type (WT) roots fed with sufficient NH4 Cl, and was induced by glutamine treatment. We generated two independent Tos17 retrotransposon insertion mutants showing reduced OsATL6 expression to determine the function of OsATL6. Compared with segregants lacking the Tos17 insertion, the OsATL6 knock-down mutant seedlings exhibited lower root glutamine content but higher glutamine concentration in the xylem sap and greater shoot growth under NH4 + -replete conditions. The transient expression of monomeric red fluorescent protein-fused OsATL6 in onion epidermal cells confirmed the tonoplast localization of OsATL6. When OsATL6 was expressed in Xenopus laevis oocytes, glutamine efflux from the cell into the acidic bath solution increased. Under sufficient NH4 + supply, OsATL6 transiently accumulated in sclerenchyma and pericycle cells, which are located adjacent to the Casparian strip, thus obstructing the apoplastic solute path, and in vascular parenchyma cells of WT roots before the peak accumulation of GS1;2 and NADH-GOGAT1 occurred. These findings suggest that OsATL6 temporarily stores excess glutamine, produced by NH4 + assimilation, in root vacuoles before it can be translocated to the shoot.

    DOI: 10.1111/tpj.15403

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  • Application of Nitrate, Ammonium, or Urea Changes the Concentrations of Ureides, Urea, Amino Acids and Other Metabolites in Xylem Sap and in the Organs of Soybean Plants (Glycine max (L.) Merr.). 国際誌

    Yuki Ono, Masashige Fukasawa, Kuni Sueyoshi, Norikuni Ohtake, Takashi Sato, Sayuri Tanabata, Ryo Toyota, Kyoko Higuchi, Akihiro Saito, Takuji Ohyama

    International journal of molecular sciences   22 ( 9 )   2021年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Soybean (Glycine max (L.) Merr.) plants form root nodules and fix atmospheric dinitrogen, while also utilizing the combined nitrogen absorbed from roots. In this study, nodulated soybean plants were supplied with 5 mM N nitrate, ammonium, or urea for 3 days, and the changes in metabolite concentrations in the xylem sap and each organ were analyzed. The ureide concentration in the xylem sap was the highest in the control plants that were supplied with an N-free nutrient solution, but nitrate and asparagine were the principal compounds in the xylem sap with nitrate treatment. The metabolite concentrations in both the xylem sap and each organ were similar between the ammonium and urea treatments. Considerable amounts of urea were present in the xylem sap and all the organs among all the treatments. Positive correlations were observed between the ureides and urea concentrations in the xylem sap as well as in the roots and leaves, although no correlations were observed between the urea and arginine concentrations, suggesting that urea may have originated from ureide degradation in soybean plants, possibly in the roots. This is the first finding of the possibility of ureide degradation to urea in the underground organs of soybean plants.

    DOI: 10.3390/ijms22094573

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  • Recent Advances in Carbon and Nitrogen Metabolism in C3 Plants. 国際誌

    Marouane Baslam, Toshiaki Mitsui, Kuni Sueyoshi, Takuji Ohyama

    International journal of molecular sciences   22 ( 1 )   2020年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    C and N are the most important essential elements constituting organic compounds in plants. The shoots and roots depend on each other by exchanging C and N through the xylem and phloem transport systems. Complex mechanisms regulate C and N metabolism to optimize plant growth, agricultural crop production, and maintenance of the agroecosystem. In this paper, we cover the recent advances in understanding C and N metabolism, regulation, and transport in plants, as well as their underlying molecular mechanisms. Special emphasis is given to the mechanisms of starch metabolism in plastids and the changes in responses to environmental stress that were previously overlooked, since these changes provide an essential store of C that fuels plant metabolism and growth. We present general insights into the system biology approaches that have expanded our understanding of core biological questions related to C and N metabolism. Finally, this review synthesizes recent advances in our understanding of the trade-off concept that links C and N status to the plant's response to microorganisms.

    DOI: 10.3390/ijms22010318

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  • Effect of the Nitrification Inhibitor 3,4-Dimethylpyrazole Phosphate on the Deep Placement of Nitrogen Fertilizers for Soybean Cultivation 招待 査読

    Soshi Hatano, Yoichi Fujita, Yoshifumi Nagumo, Norikuni Ohtake, Kuni Sueyoshi, Yoshihiko Takahashi, Takashi Sato, Sayuri Tanabata, Kyoko Higuchi, Akihiro Saito, Takuji Ohyama

    International Journal of Agronomy   2019   1 - 11   2019年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1155/2019/9724214

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  • Isolation and functional analysis of FLOWERING LOCUS T homologous gene from Vanda hybrid. 査読

    Kanokwan Panjama, Eriko Suzuki, Masahiro Otani, Masaru Nakano, Norikuni Ohtake, Takuji Ohyama, Weenun Bundithya, Kuni Sueyoshi, Soraya Ruamrungsri

    Journal of Plant Biochemistry and Biotechnology   2019年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1007/s13562-019-00487-2

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  • Effects of Different Chemical Forms of Nitrogen on the Quick and Reversible Inhibition of Soybean Nodule Growth and Nitrogen Fixation Activity. 国際誌

    Natsumi Yamashita, Sayuri Tanabata, Norikuni Ohtake, Kuni Sueyoshi, Takashi Sato, Kyoko Higuchi, Akihiro Saito, Takuji Ohyama

    Frontiers in plant science   10   131 - 131   2019年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    It has been reported that supply of nitrate to culture solution rapidly and reversibly inhibits nodule growth and nitrogen fixation activity of soybean. In this study, the effects of ammonium, urea, or glutamine on nodule growth and nitrogen fixation activity are compared with that for nitrate. Soybean plants were cultivated with a nitrogen-free nutrient solution, then 1 mM-N of nitrate, ammonium, glutamine, or urea were supplied from 12 DAP until 17 DAP. Repression of nodule growth and nitrogen fixation activity at 17 DAP were observed by ammonium, urea, and glutamine like nitrate, although the inhibitory effects were milder than nitrate. The removal of nitrogen from the culture solutions after nitrogen treatments resulted in a recovery of the nodule growth. It was found that the glutamine treatment followed by N-free cultivation gave highest nitrogen fixation activity about two times of the control. Tracer experiments with 15N and 13C were performed to evaluate the translocation of N and C to the different tissues. Culture solutions containing a 15N-labeled nitrogen source were supplied from 21 DAP, and the whole shoots were exposed to 13CO2 for 60 min on 23 DAP, and plants were harvested on 24 DAP. The percentage distribution of 15N in nodules was highest for ammonium (1.4%) followed by glutamine (0.78%), urea (0.32%) and nitrate (0.25%). The percentage distribution of 13C in the nodules was highest for the control (11.5%) followed by urea (5.8%), glutamine (2.6%), ammonium (2.3%), and nitrate (2.3%). The inhibitory effects of nitrogen compounds appeared to be related to a decrease in photoassimilate partitioning in the nodules, rather than 15N transport into the nodules. The free amino acid concentrations after nitrogen treatments were increased in the nodules and leaves by nitrate, in the roots by ammonium, in the stems by urea, and the roots, stems, and leaves by glutamine treatment. The concentrations of asparagine, aspartate, and glutamine were increased after nitrogen treatments. By the long-term supply of nitrogen for 2-weeks, nitrate significantly increased the lateral roots and leaf growth. The long-term supply of urea and glutamine also promoted the lateral roots and leaf growth, but ammonium suppressed them.

    DOI: 10.3389/fpls.2019.00131

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  • Identifying N sources that affect N uptake and assimilation in Vanda hybrid using 15N tracers

    Kanokwan Panjama, Takuji Ohyama, Norikuni Ohtake, Takashi Sato, Nuttha Potapohn, Kuni Sueyoshi, Soraya Ruamrungsri

    Horticulture, Environment, and BIotechnology   59   805 - 813   2018年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1007/s13580-018-0071-6

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  • Transcriptome and metabolome analyses reveal that nitrate strongly promotes nitrogen and carbon metabolism in soybean roots, but tends to repress it in nodules 査読

    Shinji Ishikawa, Yuki Ono, Norikuni Ohtake, Kuni Sueyoshi, Sayuri Tanabata, Takuji Ohyama

    Plants   7 ( 2 )   2018年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MDPI AG  

    Leguminous plants form root nodules with rhizobia that fix atmospheric dinitrogen (N2) for the nitrogen (N) nutrient. Combined nitrogen sources, particular nitrate, severely repress nodule growth and nitrogen fixation activity in soybeans (Glycine max [L.] Merr.). A microarray-based transcriptome analysis and the metabolome analysis were carried out for the roots and nodules of hydroponically grown soybean plants treated with 5 mM of nitrate for 24 h and compared with control without nitrate. Gene expression ratios of nitrate vs. the control were highly enhanced for those probesets related to nitrate transport and assimilation and carbon metabolism in the roots, but much less so in the nodules, except for the nitrate transport and asparagine synthetase. From the metabolome analysis, the concentration ratios of metabolites for the nitrate treatment vs. the control indicated that most of the amino acids, phosphorous-compounds and organic acids in roots were increased about twofold in the roots, whereas in the nodules most of the concentrations of the amino acids, P-compounds and organic acids were decreased while asparagine increased exceptionally. These results may support the hypothesis that nitrate primarily promotes nitrogen and carbon metabolism in the roots, but mainly represses this metabolism in the nodules.

    DOI: 10.3390/plants7020032

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  • Uptake and translocation of nitrogen in patumma (Curcuma alismatifolia) by leaves or root 査読

    Chamaiporn Anuwong, Takuji Ohyama, Kuni Sueyoshi, Norikuni Ohtake, Takashi Sato, Soraya Ruamrungsri

    JOURNAL OF PLANT NUTRITION   40 ( 8 )   1204 - 1212   2017年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS INC  

    An adequate supply of nitrogen (N) is important for patumma growth and flower quality. This study aimed to compare the uptake and translocation of N by foliar and root application. Fertilization with (15) nitrate (NO3)-N via roots or leaves was carried out at four stages, at the 1st to 4th fully expanded leaf (FEL) stages, and the plants were sampled at each successive stage. The uptake and translocation of N-15 from foliar or root applications showed relatively similar patterns at all stages. Although the N fertilizer utilization rate by roots was higher than that via leaves, the foliar application stimulated reproductive growth by earlier flowering. The N supplied at the 1st FEL and the 2nd FEL was utilized mainly in leaves, whereas supplying N at the 3rd and 4th FEL promoted flower quality. Fertilizer application method and stage of application influence the utilization rate and translocation of N to the sink organs.

    DOI: 10.1080/01904167.2016.1264597

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  • Characterisation of proteins in the storage organs of Curcuma alismatifolia Gagnep. 査読

    Chamaiporn Anuwong, Norikuni Ohtake, Kuni Sueyoshi, Soraya Ruamrungsri, Takuji Ohyama

    JOURNAL OF HORTICULTURAL SCIENCE & BIOTECHNOLOGY   89 ( 5 )   501 - 507   2014年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:HEADLEY BROTHERS LTD  

    Curcuma alismatifolia rhizomes are exported to world markets for ornamental use. Rhizome size and the number of storage roots affected shoot sprouting, plant growth, and inflorescence yield and quality, indicating an important role for storage food reserves such as the carbohydrates and proteins in these organs. An understanding of the proteins used for growth and development in rhizomes and in storage roots is lacking, so changes in the storage proteins in these organs were investigated at the planting stage, the sprouting stage, the second-leaf stage, the flowering stage, and the dormancy stage. The protein concentrations in both these organs decreased rapidly until the flowering stage in order to support the initial growth of roots and shoots. The intensity of Coomassie Brilliant Blue staining of tissues was stronger in rhizomes than in storage roots. Protein staining was present in the cytosol and in the cell walls in rhizomes, and was intense in small particle-like protein bodies in the cytosol. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles showed that peptides of 10.6 kDa and 12.0 kDa were the two major soluble proteins in rhizomes and were different from the proteins in storage roots. The 10.6 kDa and 12.0 kDa bands contained five peptides and one peptide, respectively, when separated by 2D-PAGE. The amino acid sequences of these six peptides were not homologous to any known proteins using BLASTP or FASTA analysis. The N-terminal amino acid sequences of these two polypeptides did not start with methionine, indicating that they had undergone posttranslational modification. Knowledge of the major storage proteins in the rhizomes and storage roots of C. alismatifolia may provide information to improve N-fertilisation.

    DOI: 10.1080/14620316.2014.11513112

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  • Effect of Nitrate on Nodule and Root Growth of Soybean (Glycine max ( L.) Merr.) 査読

    Akinori Saito, Sayuri Tanabata, Takanari Tanabata, Seiya Tajima, Manabu Ueno, Shinji Ishikawa, Norikuni Ohtake, Kuni Sueyoshi, Takuji Ohyama

    INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES   15 ( 3 )   4464 - 4480   2014年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MDPI AG  

    The application of combined nitrogen, especially nitrate, to soybean plants is known to strongly inhibit nodule formation, growth and nitrogen fixation. In the present study, we measured the effects of supplying 5 mM nitrate on the growth of nodules, primary root, and lateral roots under light at 28 degrees C or dark at 18 degrees C conditions. Photographs of the nodulated roots were periodically taken by a digital camera at 1-h intervals, and the size of the nodules was measured with newly developed computer software. Nodule growth was depressed approximately 7 h after the addition of nitrate under light conditions. The nodule growth rate under dark conditions was almost half that under light conditions, and nodule growth was further suppressed by the addition of 5 mM nitrate. Similar results were observed for the extending growth rate of the primary root as those for nodule growth supplied with 5 mM nitrate under light/dark conditions. In contrast, the growth of lateral roots was promoted by the addition of 5 mM nitrate. The 2D-PAGE profiles of nodule protein showed similar patterns between the 0 and 5 mM nitrate treatments, which suggested that metabolic integrity may be maintained with the 5 mM nitrate treatment. Further studies are required to confirm whether light or temperature condition may give the primary effect on the growth of nodules and roots.

    DOI: 10.3390/ijms15034464

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  • Quantitative analysis of the initial transport of fixed nitrogen in nodulated soybean plants using N-15 as a tracer 査読

    Nguyen Van Phi Hung, Shiori Watanabe, Shinji Ishikawa, Norikuni Ohtake, Kuni Sueyoshi, Takashi Sato, Satomi Ishii, Shu Fujimaki, Takuji Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   59 ( 6 )   888 - 895   2013年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    The quantitative analysis of the initial transport of fixed isotope 15-nitrogen (N-15) in intact nodulated soybean plants (Glycine max [L.] Merr. cv. Williams) was investigated at the vegetative stage (36 days after planting, DAP) and pod-filling stage (91 DAP) by the N-15 pulse-chase experiment. The nodulated roots were exposed to N-2 gas labeled with a stable isotope N-15 for 1h, followed by 0, 1, 3 and 7h of exposure with normal air. Plant roots and shoots were separated into three sections (basal, middle and distal parts) with the same length of the main stem or primary root. Approximately 80 and 92% of fixed N was distributed in the basal part of the nodulated roots at the vegetative and pod-filling stages by the end of 1h of N-15(2) exposure, respectively. In addition, about 90% of fixed N-15 was retained in the nodules and 10% was exported to root and shoot after 1 h of N-15(2) exposure at 91 DAP. The percentage distribution of N-15 in the nodules at the pod-filling stage decreased from 90% to 7% during the 7h of the chase period, and increased in the roots (14%), stems (54%), leaves (12%), pods (10%) and seeds (4%). The N-15 distribution was negligible in the distal root segment, suggesting that N fixation activity was negligible and recycling fixed N from the shoot to the roots was very low in the initially short time of the experiment.

    DOI: 10.1080/00380768.2013.838742

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  • Site of free amino acid accumulation in tulip (Tulipa gesneriana) roots during autumn and winter 査読

    Misaki Yamamoto, Toshitaka Ishida, Ryota Tanemura, Shinji Ishikawa, Norikuni Ohtake, Kuni Sueyoshi, Takuji Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   58 ( 2 )   183 - 190   2012年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    Tulip (Tulipa gesneriana, cultivar Oxford) bulbs were planted in October and grown in culture solution in a glasshouse under natural conditions. One group was cultivated in a solution containing nitrogen (N) (+N treatment: nitrate-N (NO3--N) 12.0, ammonium-N (NH4+-N) 3.0 mg L-1). Another group was cultivated in a nitrogen-free solution (-N treatment). In the first experiment, the transverse accumulation of free amino acids in the cortex and stele of the tulip roots was measured. In the cortex, the total concentration of free amino acids amounted to 39.4 mg Ng(-1) dry weight (DW) in the +N treatment, but only 5.3 mg Ng(-1) DW in the -N treatment. In the stele, the concentration was 14.4 and 5.8 mg Ng(-1) DW, respectively. Glutamine was the predominant amino acid in the cortex in the +N treatment, reaching 18.1 mg Ng(-1) DW. The concentrations of asparagine and arginine were also higher after the +N than -N treatment. On the other hand, the concentration of 4-methyleneglutamine was not so different between the treatments in either the cortex or stele. In the second experiment, the accumulation of free amino acids at five vertical positions from the base to root tip was analyzed. The total concentration of free amino acids was not so different among the positions. According to statistics, the concentrations of glutamine and asparagine were relatively constant between the +/-N treatments. In the -N treatment, the concentration of 4-methyleneglutamine gradually decreased from the base toward the root tip, and it tended to be higher in the -N treatment than that in the +N treatment.

    DOI: 10.1080/00380768.2012.669352

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  • Identification of 14-3-3 proteins as a target of ATL31 ubiquitin ligase, a regulator of the C/N response in Arabidopsis 査読

    Takeo Sato, Shugo Maekawa, Shigetaka Yasuda, Yukie Domeki, Kuni Sueyoshi, Masayuki Fujiwara, Yoichiro Fukao, Derek B. Goto, Junji Yamaguchi

    PLANT JOURNAL   68 ( 1 )   137 - 146   2011年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    The balance between carbon (C) and nitrogen (N) availability is an important determinant for various phases of plant growth; however, the detailed mechanisms regulating the C/N response are not well understood. We previously described two related ubiquitin ligases, ATL31 and ATL6, that function in the C/N response in Arabidopsis thaliana. Here, we used FLAG tag affinity purification and MS analysis to identify proteins targeted by ATL31, and thus likely to be involved in regulating the phase transition checkpoint based on C/N status. This analysis revealed that 14-3-3 proteins were associated with ATL31, and one of these, 14-3-3 chi, was selected for detailed characterization. The interaction between ATL31 and 14-3-3 chi was confirmed by yeast two-hybrid and co-immunoprecipitation analyses. In vitro assays showed that ubiquitination of 14-3-3 chi is catalyzed by ATL31. Degradation of 14-3-3 chi in vivo was shown to be correlated with ATL31 activity, and to occur in a proteasome-dependent manner. Furthermore, 14-3-3 protein accumulation was induced by a shift to high-C/N stress conditions in Arabidopsis seedlings, and this regulated response required both ATL31 and ATL6. It was also shown that over-expression of 14-3-3 chi leads to hypersensitivity of Arabidopsis seedlings to C/N stress conditions. These results indicate that ATL31 targets and ubiquitinates 14-3-3 proteins for degradation via the ubiquitin-proteasome system during the response to cellular C/N status.

    DOI: 10.1111/j.1365-313X.2011.04673.x

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  • Nitrogen and carbon translocation and distribution in curcuma alismatifolia gagnep. by labelling experiment with 15N and 13C 査読

    T. Khuankaew, S. Ito, N. Ohtake, K. Sueyoshi, T. Ohyama, T. Sato, S. Ruamrungsri

    Acta Horticulturae   886   131 - 138   2011年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:International Society for Horticultural Science  

    Curcuma alismatifolia Gagnep. rhizomes were planted on 15 May 2007. At 10 weeks after planting, at the termination of the new leaf formation stage, the plants were supplied with 15N-labeled culture solution (3.81 mM NH4 + + 8.96 mM NO3 -: 10.6 atom% 15N) twice at 3 and 1 days before 13CO2 feeding. Then 13C-labeled 13CO2 (99 atom% 13C) was exposed to the shoot. The plants were harvested at four different stages of growth. The 15N and 13C abundance in each part were determined to investigate the characteristics of N and C translocation and distribution in curcuma. The total dry weights of the plant increased continuously during the termination of the new leaf formation stage (stage 1), the flowering stage (stage 2) and the rhizome storage period (stage 3) and especially rapidly increased during stage 2 to stage 3. The DW was relatively constant thereafter until the start of the dormancy period (stage 4). The total N content of the plant rapidly increased after stage 2 until stage 4. At stage 1, the total N was distributed mostly to the old storage roots, leaves and old rhizome just as the dry weight. At stage 2, the dry weights of new organs were mostly increased as well as the N content. At stage 3 and stage 4, the total N content of the new storage organs (new rhizomes and new storage roots) was highly increased, particularly in the new rhizome. The old rhizome and the old storage roots gave very low 15N and 13C in abundance. At stage 1, leaves are the main N sink and about 5.17 mg labeled N (LN) assimilated to this organ. In this stage, translocation of the newly assimilated N to the new rhizome already started. At stage 2, the distribution of LN was increased in flower and flower stalk but it was less than in the leaves that are also the main sink of N organ. At stage 3 the distribution of LN to the aboveground part organs (leaves, flowers and flower stalks) was decreased. This contrasted to the underground part organs (new rhizome, new storage roots and fibrous roots) where LN increased especially in the new rhizome. At stage 4, the 15N was mostly distributed to the storage organ (new rhizome and new storage roots) about 8.85 mg LN and 3.59 mg LN, respectively.

    DOI: 10.17660/ActaHortic.2011.886.16

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  • Effect of NPK levels on growth and bulb quality of some geophytes in substrate culture 査読

    S. Ruamrungsri, W. Bundithya, N. Potapohn, N. Ohtake, K. Sueyoshi, T. Ohyama

    Acta Horticulturae   886   213 - 218   2011年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:International Society for Horticultural Science  

    Freesia corms were grown in substrate culture using sand: rice husk-charcoal: coconut fiber at the ratio of 1:1:1. The optimum levels of NPK in the nutrient solution were studied using different combinations of nitrogen at 100 and 200 mg/L, phosphorus at 50 and 100 mg/L combined with potassium at 100, 200 and 300 mg/L. The results showed that the minimum level of NPK at 100:50:200 mg/L was good enough for growing Freesia in substrate culture. Tubers of Gloriosa rothschildiana (10-25 g FW) were grown in substrate culture. The -N, -P, -K, -Mg and -Ca treatments compared with complete nutrient formula were supplied to the tubers. The results showed that nutrient deficiency reduced plant height, number of nodes, tuber fresh weight and tuber length. The optimum level of NPK was further studied by supplying tubers with different combinations of nitrogen at 100, 200 and 300 mg/L , phosphorus at 50 and 100 mg/L combined with potassium at 100, 200 and 300 mg/L. The results showed that the NPK combination at 200:100:200 mg/L was optimal for growing Gloriosa rothschildiana in substrate culture.

    DOI: 10.17660/ActaHortic.2011.886.29

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  • Assimilation and translocation of nitrogen and carbon in Curcuma alismatifolia Gagnep 査読

    T. Khuankaew, S. Ruamrungsri, S. Ito, T. Sato, N. Ohtake, K. Sueyoshi, T. Ohyama

    Plant Biology   12 ( 3 )   414 - 423   2010年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Curcuma or Siam tulip (Curcuma alismatifolia Gagnep.) is an ornamental flowering plant with two underground storage organs, rhizomes and storage roots. Characteristics of N and C assimilation and transport in curcuma were investigated. The plants were treated with 15NH4+ + 15NO3- and 13CO2 at 10, 13 or 21 weeks after planting. Plants were sampled at several stages up to 32 weeks. The C stored in old storage roots was used rapidly during the first 10 weeks
    after which N stored in old rhizomes and old storage roots were used. The daily gain in C depending on photosynthesis was remarkably high between 10 and 21 weeks. However, the daily gain in N was relatively constant throughout the growth period. The 15N absorbed at 10 weeks was initially accumulated in leaves and roots, but some was transported to flowering organs at 13 weeks. At harvest, 41% of 15N was recovered in new rhizomes and 17% in new storage roots. After 13CO2 exposure at 10 and 13 weeks, the distribution of 13C among organs was relatively constant in subsequent stages. When given 13CO2 at 21 weeks, a large amount of labelled C was recovered in new storage roots and new rhizomes at harvest. Both new rhizomes and new storage roots stored N and C, however, rhizomes played a more important role in supplying N, while storage roots provided C. © 2009 German Botanical Society and The Royal Botanical Society of the Netherlands.

    DOI: 10.1111/j.1438-8677.2009.00229.x

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  • Carbon and nitrogen transport during grain filling in rice under high-temperature conditions 査読

    S. Ito, T. Hara, Y. Kawanami, T. Watanabe, K. Thiraporn, N. Ohtake, K. Sueyoshi, T. Mitsui, T. Fukuyama, Y. Takahashi, T. Sato, A. Sato, T. Ohyama

    Journal of Agronomy and Crop Science   195 ( 5 )   368 - 376   2009年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Rice (Oryza sativa cv. Koshihikari) seedlings were grown in a sandy dune soil in pots with a basal dressing of N (0.5 g N), P and K. Two N treatments were applied, a +N treatment in which a top dressing of 15N-labeled 0.5 g N was supplied on July 20 and a -N treatment in which no additional fertilizer was supplied. During the grain-filling stage from August 6 to 13, plants were subjected to one of three temperature treatments
    controlled low temperature, LT (daynight 2823 °C), controlled high temperature, HT (3530 °C) and uncontrolled glasshouse temperature, UT (daynight averages, 3826 °C). All plants were then exposed to 13CO2 for 1 h on August 11 in a growth chamber at 25 °C. On August 13, all plants were harvested and the 13C and 15N abundances and starch and sugar concentrations in the ears, shoots and roots were determined. The 13C content of the ear was lower in UT than in LT irrespective of the +N or -N treatment. The translocation of 15N to the ears was also slightly depressed in UT compared with LT. Under high-temperature conditions (HT and UT), the starch content per plant was reduced for -N, but for +N, it was not significantly different among the temperature treatments. A high accumulation of sucrose was observed in all plant parts under UT conditions. It is suggested that extreme high day temperatures during the grain-filling period may reduce starch synthesis in the grains and, especially so under N-deficient conditions. High temperatures also induce an accumulation of sucrose and a decrease in carbon and nitrogen transport from the shoots to the ears via the phloem. © 2009 Blackwell Verlag GmbH.

    DOI: 10.1111/j.1439-037X.2009.00376.x

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  • Rapid quantification of cyanamide by ultra-high-pressure liquid chromatography in fertilizer, soil or plant samples 査読

    Yoshifumi Nagumo, Kazuya Tanaka, Kaushal Tewari, Khwankaew Thiraporn, Toru Tsuchida, Toshimitsu Honma, Norikuni Ohtake, Kuni Sueyoshi, Yoshihiko Takahashi, Takuji Ohyama

    JOURNAL OF CHROMATOGRAPHY A   1216 ( 29 )   5614 - 5618   2009年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    A rapid and simple method for determination of cyanamide in fertilizer, soil and plants has been developed. In this method, cyanamide is extracted with 2% acetic acid and the extract separated by centrifugation. It is then purified by passing through a membrane filter. The extract was derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl-carbamate and the derivatized compound separated by ultra-high-pressure liquid chromatography. it is then detected with a UV detector at 260 nm by the same method as is used for amino acid analysis. The proposed method is fast, simple and cheap and also has good selectivity and sensitivity for the determination of cyanamide in a wide range of biotic and abiotic materials. (C) 2009 Elsevier B.V. All rights reserved.

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  • Two-component high-affinity nitrate transport system in barley: Membrane localization, protein expression in roots and a direct protein-protein interaction 査読

    Shinji Ishikawa, Yuka Ito, Yuki Sato, Yuka Fukaya, Misa Takahashi, Hiromichi Morikawa, Norikuni Ohtake, Takuji Ohyama, Kuni Sueyoshi

    PLANT BIOTECHNOLOGY   26 ( 2 )   197 - 205   2009年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC PLANT CELL & MOLECULAR BIOL  

    A high affinity transport system (HATS) for nitrate in plants is operated by a two-component NRT2/NAR2 transport system. However, the regulation and localization of NRT2 and NAR2 at protein level are largely unknown and especially so in crop plant species. In this study with barley (Hordeum vulgare), membrane localization, protein expression in the roots, and a direct protein-protein interaction of HvNRT2 and HvNAR2 proteins were investigated. Immunochemical analysis showed that both HvNRT2 and HvNAR2 proteins were co-localized in the plasma membrane of the roots. Expression of HvNRT2 and HvNAR2 proteins was more strongly induced by treatment with higher concentrations of external nitrate, while HATS activity and transcripts for HvNRT2 and HvNAR2 were markedly repressed. An affinity Column binding analysis using recombinant proteins suggests that the C-terminus of HvNRT2.1 is possibly involved in its binding to the HvNAR2.3 central region and that the Ser463 present in the HvNRT2.1 C-terminus plays a role in the binding ability.

    DOI: 10.5511/plantbiotechnology.26.197

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  • Nitrogen Fixation and Translocation in Young Sugarcane (Saccharum officinarum L.) Plants Associated with Endophytic Nitrogen-Fixing Bacteria 査読

    Atsushi Momose, Norikuni Ohtake, Kuni Sueyoshi, Takashi Sato, Yasuhiro Nakanishi, Shoichiro Akao, Takuji Ohyama

    MICROBES AND ENVIRONMENTS   24 ( 3 )   224 - 230   2009年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC MICROBIAL ECOLOGY, DEPT BIORESOURCE SCIENCE  

    The tracer (15)N(2) was used to investigate sites of N(2) fixation and the possible translocation of the fixed N. Young sugarcane plants (Saccharum officinarum L.) from a stem cutting were exposed to (15)N(2)-labeled air in a 500 mL plastic cylinder. Plants fed (15)N(2) for 7 days were grown in normal air for a further chase period. After 21 days, about half of the N originating in the stem cutting had been transported to the shoot and roots, suggesting that the cutting played a role in supplying N for growth. After 3 days of feeding, the percentage of N derived from (15)N(2) was higher in the roots (2.22%) and stem cutting (0.271%) than the shoot (0.027%). Most of the fixed N was distributed in the 80% ethanol-insoluble fractions in each plant part, and the (15)N fixed either in the roots or in the stem cutting remained there and was not appreciably transported to the shoot. The results were quite different from the fate of fixed N in soybean nodules, which is rapidly transported from nodules to roots and shoots.

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  • Isolation and characterization of a novel Arabidopsis thaliana mutant that shows low nitrate concentration in shoots 査読

    Hiroki Ishibashi, Kimihiro Konno, Tahei Kawachi, Hisatomi Harada, Takashi Sato, Norikuni Ohtake, Takuji Ohyama, Kuni Sueyoshi

    SOIL SCIENCE AND PLANT NUTRITION   54 ( 1 )   103 - 107   2008年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    A novel Arabidopsis thaliana mutant that shows a low concentration of nitrate in shoots was isolated from an ethyl-methanesulfonate-mutagenized M-2 population. In the mutant, the nitrate concentration in the shoots was reduced to 45-61% of that in the wild-type plants when plants were grown hydroponically at a different range of external nitrate. When grown in solid media containing 5 mmol L-1 nitrate, the fresh weights of shoots and roots in the mutant were approximately 62% and 67%, respectively, compared with those of the wild type. The mutant did not show any difference in in vitro nitrate reductase activity from wild-type plants and net nitrate uptake in the mutant was 22% lower than that in wild-type plants. Genetic mapping analysis revealed that the mutation was mapped near the bottom of chromosome 1.

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  • Absorption and translocation of nitrogen in cucumber (Cucumis sativus L.) plants using the N-15 tracer technique 査読

    Ryota Tanemura, Hiroshi Kurashima, Norikuni Ohtake, Kuni Sueyoshi, Takuji Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   54 ( 1 )   108 - 117   2008年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    Cucumber (Cucumis sativus L.) plants were grown with hydroponics in a greenhouse and the absorption and translocation of nitrogen in the plants were investigated after the first fruit harvest using N-15-labeled nitrogen. The N-15-labeled nutrient solution, which contained 13 mmol L-1 nitrate, was supplied for 7 days from first fruit harvest on 21 September, 2005. Following 7 days of N-15 treatment, these plants received the same composition of non-labeled nitrogen for 16 days (chase period). Commercial-sized fruits, approximately 20 cm in length, were harvested every day. The whole plants were harvested three times at the beginning (21 September) and at the end (28 September) of the N-15 treatment period, and at the end of the chase period on 14 October. The average nitrogen absorption rate was 196 mg plant(-1) per day during the N-15 treatment period and 181 mg plant(-1) per day during the chase period. The labeled nitrogen was detected in the fruits within 1 day of the N-15 treatment, and the percentage of nitrogen from labeled nitrogen in the harvested fruits increased up to approximately 50% at the end of the N-15 treatment period. The percentage of labeled nitrogen in the fruits quickly decreased after changing from N-15 solution to the non-labeled conditions during the chase period. At the end of the N-15 treatment period on 28 September, the percentage distribution of labeled nitrogen was 46% in the leaves, 24% in the harvested fruits, 13% in the stems, 9% in the roots and 9% in the small fruits. After 16 days of chase period, the distribution of labeled nitrogen decreased in the leaves (33%), stems (6.5%), roots (3.6%) and small fruits (1%), but 56% of labeled nitrogen was distributed in the harvested fruits. Based on the result obtained, the nitrate absorbed from the medium is rapidly transported to young fruits as well as to leaves and stems. The nitrogen once assimilated or stored in the leaves, stems and roots is gradually redistributed to the fruits for supporting their rapid growth. To maximize the yield of cucumber fruits, it is necessary to supply sufficient levels of nitrogen continuously after the first harvest.

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  • The autoregulation of nodulation mechanism is related to leaf development 査読

    Sayuri Ito, Taichi Kato, Norikuni Ohtake, Kuni Sueyoshi, Takuji Ohyama

    PLANT AND CELL PHYSIOLOGY   49 ( 1 )   121 - 125   2008年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    To understand the autoregulation of nodulation (AON) system, in which leguminous plants control the nodule number, we examined the details of the characteristics of hypernodulation soybean mutants NOD1-3 and NOD3-7. A microscopic study showed that NOD1-3 and NOD3-7 produced small-size leaves due to the smaller number of leaf cells, compared with the Williams parent. These phenotypes were not affected by inoculation with bradyrhizobia or nitrate supply. The AON signaling might be related to the control system of leaf cell proliferation. This hypothesis was strongly supported by the finding that activation of AON in wild types by inoculation leads to an increase in the cell number of leaves.

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  • COMPARISON OF LEAF GROWTH OF HYPERNODULATION SOYBEAN MUTANTS, NOD1-3, NOD2-4 AND NOD3-7, WITH THEIR PARENT CV. WILLIAMS 査読

    S. Ito, N. Ohtake, K. Sueyoshi, T. Ohyama

    BIOLOGICAL NITROGEN FIXATION: TOWARDS POVERTY ALLEVIATION THROUGH SUSTAINABLE AGRICULTURE   42   193 - +   2008年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:SPRINGER  

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  • Changes of carbohydrate contents in curcuma alismatifolia Gagnep. Affected by red light 査読

    A. Chidburee, W. Bundhitaya, C. Suwanthada, N. Ohtake, K. Sueyoshi, T. Ohyama, S. Ruamrungsri

    Acta Horticulturae   788   131 - 136   2008年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:International Society for Horticultural Science  

    The effects of red light on carbohydrate content in Curcuma alismatifolia Gagnep. were studied. Plants grown under two artificial light sources: T1) red light fluorescent lamp (Philips TLD 36W/15
    632-660 nm) and T2) cool daylight source (Philips TLD 36W/865
    405-812 nm) at 27±2°C, 70-80% RH and 60 μmol m-2s-1 PAR of light intensity compared with growing under natural condition (T3). Each plant were supply with 100 ml of nutrient solution comprised of 200 mg N, 50 mg P, 200 mg K, 65 mg Ca, 20 mg Mg, 0.22 mg B, 0.54 mg Mn, 0.26 mg Zn, 0.04 mg Mo and 0.45 mg Fe per litre, three times a week. The starch content in new rhizome of plants growing under red light was the lowest. Total soluble sugar, fructose, glucose and sucrose of plants growing under natural light were the highest. The contents of starch and total soluble sugar in storage roots of plant grown under natural light were the highest.

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  • MECHANISM OF QUICK AND REVERSIBLE INHIBITION OF SOYBEAN NODULE GROWTH AND NITROGEN FIXATION ACTIVITY BY NITRATE AND ITS METABOLITES 査読

    T. Ohyama, A. Yamazaki, N. Yamashita, T. Kimura, S. Ito, N. Ohtake, K. Sueyoshi

    BIOLOGICAL NITROGEN FIXATION: TOWARDS POVERTY ALLEVIATION THROUGH SUSTAINABLE AGRICULTURE   42   89 - +   2008年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:SPRINGER  

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  • Analysis of the nitrogen nutrition of soybean plants with deep placement of coated urea and lime nitrogen 査読

    Kaushal Tewari, Takanori Sato, Mayumi Abiko, Norikuni Ohtake, Kuni Sueyoshi, Yoshihiko Takahashi, Yoshifumi Nagumo, Toru Tutida, Takuji Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   53 ( 6 )   772 - 781   2007年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    In the present study, the effect and utilization of deep placement N-15-labeled coated urea (CU) and lime nitrogen (LN) were investigated at various growth stages of soybean (Glycine max [L.]). Soybean (cv. Enrei) plants were planted in a rotated paddy field in Niigata, Japan. N-15-labeled CU or LN (100 kg N ha(-1)) was supplied separately at a depth of 20 cm just below the seeding line. Deep placement of LN and CU significantly enhanced the dry weight and N content of the plants compared with the control treatment (Cont) without deep placement. As a result, the seed yield per plant in CU (67.2 g) and LN (70.6 g) was much higher than the Cont (37.1 g). N-15 analysis of plants showed that the pattern of labeled N absorption tended to be lower with LN than CU at the R3 and R5 stages, but the recovery rate at R7 was higher in LN (70%) than CU (61%). Combining the N-15 analysis with a relative ureide analysis, the N derived from nitrogen fixation (Ndfa), from fertilizer (Ndff) and from soil (Ndfs) were evaluated. At the R7 stage, the amount of Ndfa was higher in CU (32.1 g m(-2)) and LN (31.1 g m(-2)) than in Cont (21.4 g m(-2)). This positive response for N-2 fixation by the deep placement of CU and LN may result from the continuous supply of N from the lower parts of the roots, which promotes shoot growth and extends the photosynthetic activity of the green leaves, resulting in the promotion of nitrogen fixation and seed yield.

    DOI: 10.1111/j.1747-0765.2007.00194.x

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  • Characteristics of initial growth of hypernodulation soybean mutants, NOD1-3, NOD2-4 and NOD3-7, affected by inoculation of bradyrhizobia and nitrate supply 査読

    Sayuri Ito, Norikuni Ohtake, Kuni Sueyoshi, Takuji Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   53 ( 1 )   66 - 71   2007年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    The characteristics of plant growth of hypernodulation soybean mutant lines NOD1-3, NOD2-4 and NOD3-7 were compared with their parent cv. Williams. The plants were grown using hydroponics in an illuminated growth chamber with or without seed inoculation of bradyrhizobia, and in the absence or presence of nitrate in the culture solution. When the plants were grown without inoculation, the total dry weight of all mutant lines was not different statistically from Williams, both in the absence and presence of nitrate. When they were grown with inoculation of bradyrhizobia, however, the total dry weight of each mutant line was significantly lower than that of Williams, both in the absence and presence of nitrate. These results indicate that the reduced accumulation of total dry matter of hypernodulation mutant lines compared with the wild type may be a secondary effect passively resulting from the large number of nodules, while the hypernodulation trait is a primary effect of the mutated gene. When the plants were grown with inoculation, the nodule number was decreased by the presence of nitrate in Williams, NOD1-3 and NOD2-4, but not in NOD3-7. NOD3-7 may be the most tolerant to nitrate inhibition of nodulation among the NOD mutant lines. In contrast, leaf growth of NOD3-7 and NOD1-3 was different from the wild type; the expanded leaf was smaller, but the leaf emergence rate was faster compared with Williams under all conditions. This indicates that NOD3-7 and NOD1-3 might decrease the ability for leaf expansion or may have a faster leaf emergence rate.

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  • Effect of deep placement of slow-release fertilizer (Lime nitrogen) applied at different rates on growth, N-2 fixation and yield of soya bean (Glycine max [L.] Merr.) 査読

    T. Kaushal, M. Onda, S. Ito, A. Yamazaki, H. Fujikake, N. Ohtake, K. Sueyoshi, Y. Takahashi, T. Ohyama

    JOURNAL OF AGRONOMY AND CROP SCIENCE   192 ( 6 )   417 - 426   2006年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    A new fertilization method with deep placement of slow-release N fertilizers, such as coated urea and lime nitrogen (LN) (calcium cyanamide) at 20 cm depth was found to promote soy bean seed yield. In the present study, the effect of deep placement of LN was investigated on different parameters such as growth, N accumulation, NZ fixation activity and yield of soy bean by applying LN at different rates in the rotated paddy field of Niigata, Japan. In addition to the basal fertilizer, ammonium sulphate (16 kg N ha(-1)), deep placement of LN was conducted by applying various amounts such as 50 kg N ha(-1) (A50), 100 kg N ha(-1) (A100) and 200 kg N ha(-1) (A200) at 20 cm depth in separate plots. A N-15-labelled LN fertilizer was also employed for each of the above treatments to calculate N utilization from LN in separate plots. Soya bean plant growth and N-2 fixation activity were periodically analysed. Both plant growth and N accumulation were found to increase with LN treatment compared with control plants. An increase in N-2 fixation activity was found in the A100 plots. The total seed yield was the highest in the deep placement of LN with A100 (73 g per plant) compared with other treatments. The visual quality of harvested seeds also showed that A100 enhanced the quality of seeds compared with other treatments. Thus, it is suggested that N fertilization management with particular reference to optimum amount of fertilizers is important for maximum growth. N-2 fixation and enhancement of seed yield of soy bean.

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  • Effect of nitrogen supply on nitrogen and carbohydrate constituent accumulation in rhizomes and storage roots of Curcuma alismatifolia Gagnep. 査読

    Norikuni Ohtake, Soraya Ruamrungsri, Sayuri Ito, Kuni Sueyoshi, Takuji Ohyama, Pimchal Apavatjrut

    SOIL SCIENCE AND PLANT NUTRITION   52 ( 6 )   711 - 716   2006年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    Curcuma (Curcuma alismatifolia cv. Gagnep.), a tropical flowering plant known as "Siam tulip", were cultivated in a pot with vermiculite and supplied with different levels of nitrogen (N). Rhizomes with storage roots were harvested at 215 days after planting. Results indicated that a high level of N supply increased flower numbers and promoted continuous new rhizome formation, but storage root growth was depressed. The N supply to the plants increased the N concentrations both in the rhizomes and in the storage roots. The predominant nitrogenous compounds related to total N increase were proteins in the rhizomes. The N of the insoluble fraction of 80% ethanol or the N of the soluble fraction of 10% trichloroacetic acid was the predominant fraction of N that accumulated in the storage roots. A lack of N supply increased the starch concentration both in the rhizomes and in the storage roots. These results suggested that a high level of N supply to the curcuma plant increased new rhizome formation because of increased flower numbers, but depressed new storage root formation because of reduced starch accumulation.

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  • 硝酸イオンによるダイズ根粒の肥大生長と窒素固定活性の阻害機構

    大山 卓爾, 伊藤 小百合, 大竹 憲邦, 末吉 邦

    化学と生物   44 ( 11 )   752 - 759   2006年11月

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    記述言語:日本語   出版者・発行元:Japan Society for Bioscience, Biotechnology, and Agrochemistry  

    DOI: 10.1271/kagakutoseibutsu1962.44.752

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  • Comparison of the depth of placement of lime nitrogen on growth, N-2 fixation activity, seed yield and quality of soybean (Glycine max (L.) Merr.) plants 査読

    Tewari Kaushal, Masaru Onda, Sayuri Ito, Akihiko Yamazaki, Hiroyuki Fujikake, Norikuni Ohtake, Kuni Sueyoshi, Yoshihiko Takahashi, Yoshifumi Nagumo, Tohru Tsuchida, Takuji Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   52 ( 4 )   453 - 463   2006年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    We have shown in previous reports that a new fertilization method with deep placement of slow-release N fertilizers, such as coated urea and lime nitrogen (calcium cyanamide), at a depth of 20 cm promoted soybean growth and seed yield. In the present study, the effect of deep placement of lime nitrogen (which gave the highest yield in our previous experiment) was compared at various depths in a rotated paddy field at the Niigata Agricultural Research Institute. The objective of the present study was to analyze growth, N-2 fixation activity and seed yield, and to examine visually the quality of soybean seeds cultivated at different depths with the slow-release fertilizer, lime nitrogen. In addition to the conventional basal application of mixed chemical fertilizer (including ammonium sulfate 16 kg N ha(-1)) in 0-10 cm soil layers, deep placement of lime nitrogen was conducted by applying at various depths (10 cm [D10], 15 cm [D15] and 20 cm [D20]) the same amount (100 kg N ha(-1)) in separate plots. N-15-labeled lime nitrogen fertilizers were also used for each of the above treatments to calculate N absorption in relation to fertilization depth. Soybean plant growth was periodically analyzed and the quality of harvested seeds was also visually examined (hereafter referred to as "visual quality"). The total seed yield was highest in the deep placement of lime nitrogen (100 kg N ha(-1)) at D20 (69 g plant(-1)) compared with D15 (59 g plant(-1)), D10 (57 g plant(-1)) and the control (48 g plant(-1)). The visual quality of harvested seeds showed that lime nitrogen (20 cm depth and 100 kg N ha(-1)) enhanced the quality of seeds compared with the other treatments, in which the good quality seeds (hereafter referred to as "good seeds") based on dry weight were 24 g plant(-1) (control), 37 g plant(-1) (D10), 33 g plant(-1) (D15) and 45 g plant(-1) (D20). Thus, it appears that N fertilization management with particular reference to the depth of fertilizer placement is important for maximum yield and enhancement of seed quality of soybean.

    DOI: 10.1111/j.1747-0765.2006.00056.x

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  • Allocation of photosynthetic products in soybean during the early stages of nodule formation 査読

    Sayuri Ito, Norikuni Ohtake, Kuni Sueyoshi, Takuji Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   52 ( 4 )   438 - 443   2006年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    A time-course study examining the current photosynthate allocation of soybean (Glycine max [L.] Merr.) cv. Williams was conducted in relation to nodule initiation. Whole shoots were exposed to (CO2)-C-14 for 120 min and the distribution of radioactivity in each organ was determined. During the early stages of nodule formation (i.e. 4, 6 and 8 days after inoculation) the C-14 distribution to the inoculated roots did not increase when compared with uninoculated control roots. In addition, the C-14 respired by underground parts was similar in both the inoculated and the control roots. Eight days after inoculation, the accumulation of starch and sugar was similar in both inoculated and uninoculated plants. These results indicate that photosynthate allocation for nodule initiation does not increase markedly during the early stages of nodule formation. After the emergence of the nodules, photosynthate allocation to the inoculated roots gradually increased. In addition, the consumption of current photosynthate by the respiration of underground parts increased at day 12 after inoculation, but did not increase at day 8 after inoculation.

    DOI: 10.1111/j.1747-0765.2006.00051.x

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  • Determination of the uptake and utilization of nitrogen in Curcuma alismatifolia Gagnep. using N-15 isotope 査読

    S Ruamrungsri, N Ohtake, K Sueyoshi, T Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   52 ( 2 )   221 - 225   2006年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    Rhizomes of Curcuma alismatifolia Gagnep. were planted in a soil-less culture medium composed of sand: perlite mixed at a ratio of 1:1. Three groups of plants were fed with a N-15 ((NO3-)-N-15 40 mg N L-1 + (NH4+)-N-15 10 mg N L-1)-labeled culture solution over three different periods: (1.) Period 1: from root emergence (2 weeks after planting [WAP]) to the two expanded leaf stage (Stage 1: 6 WAP), (2) Period 2: from Stage 1. to the first floret opening stage (Stage 2: 9 WAP), (3) Period 3: from Stage 2 to harvest during dormancy (Stage 3: 24 WAP). Plants received the same composition of non-labeled N from 2 WAP to 24 WAP, except for the N-15-labeling period. The fourth group of plants was cultured in a nitrogen-free solution and the fifth group was cultured in a complete solution (referred to as control treatment) from planting until harvest and sampled to determine the changes in total N content. Nitrogen content and N-15 abundance were determined at different stages of growth. Average absorption of N-15 from the culture solution was 23.5 mg N plant(-1) during Period 1, 97.8 mg N plant(-1) during Period 2 and 78.7 mg N plant(-1) during Period 3. Daily N absorption rates per plant were 0.84 mg N during Period 1, 4.66 mg N during Period 2 and 0.75 mg N during Period 3. A large amount of absorbed nitrogen was translocated to the compact spike, particularly in Period 2. At harvest the nitrogen accumulation was partitioned almost equally between new rhizomes and storage roots. Most of the absorbed nitrogen in the new rhizomes and storage roots was derived from that received during Period 3.

    DOI: 10.1111/j.1747-0765.2006.00027.x

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  • Genetic engineering of nitrite reductase gene improves uptake and assimilation of nitrogen dioxide by Rhaphiolepis umbellata (Thunb.) Makino 査読

    Jun Shigeto, Sanae Yoshihara, Suaad E.H. Adam, Kuni Sueyoshi, Atsushi Sakamoto, Hiromichi Morikawa, Misa Takahashi

    Plant Biotechnology   23 ( 1 )   111 - 116   2006年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:Japanese Society for Plant Cell and Molecular Biology  

    Hypocotyl sections of Rhaphiolepis umbellata (Thunb.) Makino were transformed by Agrobacterium tumefaciens bearing a binary vector pIG121-AtNiR, which contains cDNA of the nitrite reductase gene from Arabidopsis thaliana (Atni) under the control of a modified cauliflower mosaic virus 35S promoter and chimeric hygromycin phosphotransferase gene (hph). A 4% of the hypocotyl explants transfected with Agrobacterium formed hygromycin resistant adventitious shoots, and most of them rooted upon root induction treatment. The presence and expression of the introduced transgene in putative transgenic plants (12 months after transfection) were respectively confirmed by polymerase chain reaction (PCR) analysis using primers specific to Atni and by western blot analysis using anti NiR antibody. A total of 37 transgenic plant lines were obtained. Plants 33 months after the transfection were fumigated with 200±50 ppb 15NO2 under the natural light for one week in the fumigation chamber in a confined glasshouse. The amount of total nitrogen derived from NO2 (reflecting uptake of NO2) and that of Kjeldahl nitrogen derived from NO2 (reflecting assimilation of NO2) were determined using a mass spectrometer. One of the 9 transgenic plants tested was 1.6-2.0 times higher both in the uptake and assimilation of NO2 than non-transformed wild-type ones.

    DOI: 10.5511/plantbiotechnology.23.111

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  • Long-term effect of nitrate application from lower part of roots on nodulation and N-2 fixation in upper part of roots of soybean (Glycine max (L.) Merr.) in two-layered pot experiment 査読

    H Yashima, H Fujikake, A Yamazaki, S Ito, T Sato, K Tewari, N Ohtake, K Sueyoshi, Y Takahashi, T Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   51 ( 7 )   981 - 990   2005年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    The long-term effect of the concentration and duration of application of nitrate from the lower part of soybean roots on the nodulation and nitrogen fixation in the upper part of roots was investigated using a two-layered pot system separating the upper roots growing in a vermiculite medium and the lower roots growing in a nutrient solution. Continuous absence of nitrate (hereafter referred to as "0-0 treatment"), and continuous 1 mm (1-1 treatment) and 5 mm (5-5 treatment) nitrate treatments were imposed in the lower pot from transplanting to the beginning of the maturity stage. In addition, 5 mm nitrate was supplied partially from the beginning of the pod stage till the beginning of the maturity stage (0-5 treatment) or from transplanting till the beginning of the pod stage (5-0 treatment). The values of the total plant dry weight and seed dry weight were highest in the 5-5 treatment, intermediate in the 1-1, 5-0, 0-5 treatments, and lowest in the 0-0 treatment. The values of the nodule dry weight and nitrogen fixation activity (acetylene reduction activity) were lowest in the 5-5 treatment. The value of the nodule dry weight in the upper roots was highest in the plants subjected to the 1-1 treatment and exceeded that in the 0-0 treatment. Total nitrogen fixation activity of the upper nodules per plant at the beginning of the pod stage was also highest in the 1-1 treatment. These results indicated that long-term supply of a low level of nitrate from the lower roots could promote nodulation and nitrogen fixation in the upper part of roots. Withdrawal of 5 mm nitrate after the beginning of the pod stage (5-0 treatment) markedly enhanced nodule growth and ARA per plant in the upper roots at the beginning of the maturity stage when the values of both parameters decreased in the other treatments. The nitrate concentration in the nodules attached to the upper roots was low, including the 5-5 treatment regardless of the stages of growth. This indicated that the inhibitory effect of 5 mm nitrate or promotive effect of 1 mm nitrate supplied from the lower roots was not directly controlled by nitrate itself, but was mediated by some systemic regulation, possibly by the C or/and N requirement of the whole plant.

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  • N-15 analysis of the promotive effect of deep placement of slow-release N fertilizers on growth and seed yield of soybean 査読

    T Kaushal, M Onda, S Ito, A Yamazaki, H Fujikake, N Ohtake, K Sueyoshi, Y Takahashi, T Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   51 ( 6 )   885 - 892   2005年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    It is well known that nodulation and nitrogen fixation of soybean are generally depressed by the application of N fertilizers. We previously reported that a new fertilization method consisting of deep placement of slow-release N fertilizers, such as coated urea and lime nitrogen (calcium cyanamide) at a 20 cm depth enhanced soybean growth and seed yield. In the present study, the effect of deep placement of various N-15-labeled N fertilizers (100 kg N ha(-1)) was compared in converted paddy fields located at Niigata Agricultural Research Institute, in Nagaoka. Fertilizers consisting of N-15-labeled ammonium sulfate (AS), urea (U), coated urea (CU) or lime nitrogen (LN) were applied at a 20 cm depth for nodulated soybean (cv. Enrei) and for a non-nodulated isogenic line (En1282). The plants without deep placement of N fertilizer were used as control (Cont) for both Enrei and En1282. Whole plants were sampled at maturing stage, and N-15 abundance and N concentration in each part were determined. The evaluation of the amounts of Ndfa, Ndfs and Ndff was conducted by the N-15 dilution method using En1282 as a reference plant. Also the amounts of Ndfa and Ndfs+Ndff were estimated by the N balance method for comparing Cont and the other treatments. In all the deep placement treatments, the non-nodulated line exhibited only 36-40% of total dry weight and 16-30% of total N accumulation compared with the nodulated line, due to N deficiency associated with the lack of nitrogen fixation. In both nodulated and non-nodulated soybeans, plant growth and the total amount of accumulated N showed the highest values by the application of LN, followed by CU, U, AS and Cont, respectively: The value of the seed dry weight of Enrei was highest in the LN (73 g plant(-1)), followed by CU (63 g plant(-1)), U (47 g plant(-1)), AS (37 g plant(-1)) and Cont (26 g plant(-1)) treatments. The value of Ndfa estimated by the N-15 dilution method was higher in the LN (3.6 g N plant(-1)) and CU (2.8 g N plant(-1)) treatments than in the U (2.3 g N plant(-1)) and AS (2.2 g N plant(-1)) treatments. In addition, the value of Ndfa estimated by the N-balance method was also higher in the LN (4.1 g N plant(-1)), CU (3.9 g N plant(-1)), U (2.8 g N plant(-1)) and AS (2.6 g N plant(-1)) treatments than in the Cont treatment (1.5 g N plant(-1)). The recovery rate of N fertilizers in Enrei was higher in the LN (43%) and CU (36%) treatments than in the U (21%) treatment and lowest in the AS (8.5%) treatment. These results indicate that deep placement of LN and CU is effective to improve soybean growth and seed yield. The combination of "deep placement" and "slow-release N fertilizers" is important to supply N until the maturation stage, resulting in the increase of seed yield.

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  • Analysis of promotive effects of deep placement of slow release fertilizers on growth and seed yield of soybean by 15N dilution method 査読

    T Kaushal, M Onda, S Ito, A Yamazaki, H Fujikake, N Ohtake, K Sueyoshi, Y Takahashi, T Ohyama

    Biological Nitrogen Fixation, Sustainable Agriculture and the Environment   41   287 - 288   2005年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:SPRINGER  

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  • Effect of placement of urea and coated urea fertilizers on yield and quality of soybean (Glycine max (L.) Merr.) seeds 査読

    T Kaushal, M Onda, S Ito, A Yamazaki, H Fujikake, N Ohtake, K Sueyoshi, Y Takahashi, T Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   50 ( 8 )   1245 - 1254   2004年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    The objective of the present study was to record the seed yield and to examine visually the quality of soybean seeds cultivated under different types and placements of urea fertilizers. In addition to the conventional fertilizer application (including ammonium sulfate 16 kg N ha(-1)), broadcasting (100 kg N ha(-1)) of urea (UB) and 100-d type coated urea CU-100 (CUB), and deep placement (100 kg N ha(-1)) of urea (UD) and 100-d type coated urea CU-100 (CUD) was conducted in separate plots in a paddy field converted to an upland field located at Shindori Experimental Station of Niigata University. Soybean plant growth was periodically analyzed and the quality of harvested seeds was also visually examined (hereafter referred to as "visual quality"). It was found that the deep placement treatments were more conducive to nitrogen (N,) fixation, based on the relative ureide N concentration in the xylem sap, which is a good indicator of N, fixation by soybean. Also the total seed yield was the highest in CUD (82 g plant(-1)) and UD (81 g plant(-1)), compared to the control (62 g plant(-1)), UB (68 g plant(-1)), and CUB (68 g plant(-1)). The visual quality of harvested seeds showed that CUD enhanced the quality of seeds compared to the other treatments, in which the percentage of good quality seeds, hereafter referred to as "good seeds," based on the dry weight was 51 (control), 65 (UB), 61 (CUB), 61 (UD), and 66% (CUD). In terms of diseased seeds, the percentage of turtle wrinkle and broken seed coats was found to decrease by N application compared to the control. Thus, it is suggested that N fertilization management is important for maximum yield of soybean as well as for the enhancement of seed quality.

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  • Contribution of shoots and roots to in vivo nitrate reduction in NADII-Specific nitrate reductase-deficient mutant seedlings of barley (Hordeum vulgare L.)

    S Abdel-Latif, T Kawachi, H Fujikake, N Ohtake, T Ohyama, K Sueyoshi

    SOIL SCIENCE AND PLANT NUTRITION   50 ( 4 )   527 - 535   2004年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    Barley (Hordeum vulgare L.) has two nitrate reductase (NR) isozymes, namely NADH-specific NR and NAD(P)H-bispecific NR. To determine the effect of NADH-specific NR deficiency on in vivo nitrate reduction and distribution of reduced N to shoots and roots, the N-15-incorporation model was applied to NADH-specific NR-deficient mutant (Az12) seedlings of barley. The N-deprived seedlings were treated with a nutrient solution containing 2.3 mm NO3- and 0.2 mm NO2-, and were labeled with different amounts of 15 N for 48 It under continuous illumination. In Az12, the total in vitro NR activity derived from the NAD(P)H-bispecific NR was only 10% of that of the wild type (cv. Steptoe). In Az12, the total (NO3-)-N-15 incorporation and translocation of absorbed (NO3-)-N-15 to shoots were about 25% higher than those of Steptoe. Nitrate reduction in the Az12 roots was 2 times higher than that in Steptoe during the first period (0-24 h) and 1.3 times higher during the second period (24-48 h). However, nitrate reduction in the Az12 shoots was 10 to 30% lower than that in the Steptoe shoots. As a result, nitrate markedly accumulated in the Az12 shoots. Accumulation of reduced N-15 in the Az12 roots was 2 times higher than that in the Steptoe roots, but 10% lower in the Az12 shoots than in the Steptoe shoots at the end of the experiment. Upward transport of reduced N-15 via the xylem in Az12 was nearly 2 times more active than that of Steptoe throughout the experiment. This result suggested that the derepression of the NAD(P)H-NR isozyme in the Az12 shoots could compensate for the absence of the NADH-NR isozyme. Furthermore, increased levels of root nitrate reduction seemed to make up for the limited nitrate assimilation in the Az12 shoots.

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  • Quantitative and isotopic analysis of amino acids, allantoin, and allantoic acid in soybeans by LC-MS using the atmospheric pressure chemical ionization method 査読

    N Ohtake, A Takano, S Ito, A Yamazaki, H Fujikake, K Sueyoshi, T Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   50 ( 2 )   241 - 248   2004年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    In this report, we determined whether liquid chromatography mass spectrometry (LC-MS) using the atmospheric pressure chemical ionization (APCI) method could be applied for isotopic analysis of the high polar compounds, allantoin, and allantoic acid, which are the major transport forms of N from soybean nodules. The peak areas of the ion chromatogram of authentic allantoin and allantoic acid obtained by selected ion monitoring (SIM) were proportional to the contents of the compounds. The data obtained from the ion intensity using the scan mode and the area of the ion chromatogram using the SIM mode were entirely consistent with the isotope ratio of the compounds. The allantoin and allantoic acid concentrations in the xylem sap of soybeans at various growth stages were measured by the following two methods using LC-MS; an absolute calibration method and an isotope-labeled internal standard method. The values estimated by the isotope-labeled internal standard method were more consistent with those obtained by the colorimetric method. Furthermore, amino acids could be detected under the same analytical conditions. The N-15 ratio of asparagine and aspartic acid in the non-nodulating soybean xylem sap which had been supplied with N-15-labeled nitrate could be measured. These results indicate that LC-MS (APCI) is a suitable method for isotopic analysis of high polar compounds, including allantoin and allantoic acid in plant xylem sap and tissues.

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  • 日本で広く栽培されているレタス品種におけるアグロバクテリウム法による形質転換

    中野 優, 牧野 太英, 末吉 邦

    新潟大学農学部研究報告   56 ( 2 )   59 - 66   2004年3月

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    記述言語:英語   出版者・発行元:新潟大学  

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  • Effect of deep placement of N fertilizers and different inoculation methods of bradyrhizobia on growth, N-2 fixation activity and N absorption rate of field-grown soybean plants 査読

    K Tewari, T Suganuma, H Fujikake, N Ohtake, K Sueyoshi, Y Takahashi, T Ohyama

    JOURNAL OF AGRONOMY AND CROP SCIENCE   190 ( 1 )   46 - 58   2004年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL VERLAG GMBH  

    The effects of deep placement (supplied at 20 cm depth from soil surface below plants) of 100 kg N ha(-1) of N fertilizers, urea, coated urea or calcium cyanamide (lime nitrogen) on the growth, nitrogen fixation activity, nitrogen absorption rate and seed yield of soybean (Glycine max L. Merr.) plants were examined by comparing them with control plots without deep placement of N fertilizer in sandy dune field. In addition, three different inoculation methods of bradyrhizobia were used for each N treatment: (1) transplantation of 10-day-old seedling in a paper pot with vermiculite inoculated with Bradyrhizobium japonicum USDA110, (2) direct transplantation of inoculated 10-day-old seedlings, and (3) transplantation of 10-day-old seedlings in a non-inoculated paper pot. The deep placement of N fertilizers, especially calcium cyanamide and coated urea, markedly increased the growth and total N accumulation in shoot, roots and nodules, which resulted in an increase in seed yield. Daily N-2 fixation activity and N absorption rate were estimated by relative abundance of ureide-N analysed from the concentration of N constituents (ureide-N, amide-N and nitrate-N) in root bleeding xylem sap and increase in total N accumulation in whole plants at R1, R3, R5 and R7 stages. The total amount of N-2 fixation was about 50 % higher in the plants with calcium cyanamide and coated urea deep placements compared with control plants. Deep placement of slow release fertilizers kept nodule dry weight higher in the maturing stage of seed, possibly through abundant supply of photoassimilate to the nodules by supporting leaf area and activity until late reproductive stages. The results indicate that deep placement of calcium cyanamide or coated urea enhances N-2 fixation activity, which ultimately increases the seed yield. The promotive effect was observed with the seedlings transplanted in paper pot with inoculum of bradyrhizobia within any treatments, although nodulation by indigenous rhizobia was observed in the plants transplanted with non-inoculated paper pot.

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  • Systemic and local effects of long-term application of nitrate on nodule growth and N-2 fixation in soybean (Glycine max [L.] Merr.) 査読

    H Yashima, H Fujikake, T Sato, N Ohtake, K Sueyoshi, T Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   49 ( 6 )   825 - 834   2003年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    The effect of nitrate placement on the nodule number, nodule growth, and N-2 fixation activity in soybean plant was elucidated by using a two-layered pot system separating the upper roots grown in vermiculite medium from the lower roots cultured in hydroponics. Four treatments were imposed i.e. 0/0, 0/5, 5/0, and 5/5, with the 0 or 5 mm nitrate treatment in the upper pot/lower pot, respectively. The plants were harvested at the flowering stage and initial pod setting stage. Systemic and local inhibition on nodule number did not occur in the upper nodules in vermiculite. On the other hand, systemic inhibition on the nodule growth and N-2 fixation activity in the upper pot was apparent at pod setting stage. In the lower pot where the nodules were in direct contact with 5 mm nitrate, inhibition on nodule number, nodule size, and N-2 fixation activity were conspicuous. Systemic inhibition on the number and dry weight of lower nodules was also observed. Root growth was promoted with the root part in direct contact with 5 mm nitrate, irrespective of upper or lower roots. Nitrate accumulation was observed only in the part of roots and nodules in direct contact with 5 mm nitrate. The results indicated that systemic nitrate inhibition occurred both on N-2 fixation activity and on nodule growth but not on the upper nodule number after long-term application of 5 mm nitrate. Milder inhibition on upper nodules compared with the lower nodules may be due to the rapid depletion of nitrate from the vermiculite medium compared with the culture solution in the lower pot.

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  • Photoassimilate partitioning in hypernodulation mutant of soybean (Glycine max (L.) Merr.) NOD1-3 and its parent Williams in relation to nitrate inhibition of nodule growth 査読

    H Fujikake, Y Tamura, N Ohtake, K Sueyoshi, T Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   49 ( 4 )   583 - 590   2003年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    Nodule growth of a hypernodulating soybean (Glycine max (L.) Merr.) mutant line NOD1-3 was compared to that of its wild-type parent cv. Williams from 14 to 18 days after planting (DAP) in the absence of nitrate treatment (hereafter referred to as "0 mm treatment") or with 5 mm nitrate treatment. The growth rate determined by increase in the diameter of the nodules was relatively lower in the mutant NOD1-3 than that of the parent Williams under nitrogen-free conditions (0 mm nitrate). The inhibition of nodule growth by 5 mm nitrate started at 1 d after the onset of the nitrate treatment in Williams, while the inhibition did not occur before the application of the nitrate treatment for 2 d in NOD1-3. The nodule growth was completely inhibited after 2 d in Williams and after 3 d in NOD1-3 during the 5 mm nitrate treatment period. After 4 d of 5 mm nitrate treatment, the nodule dry weight decreased by 22% in NOD1-3 and by 58% in Williams, respectively. The treatment with 5 mm nitrate decreased the acetylene reduction activity (ARA) in NOD1-3 by 60% per plant and by 50% per nodule g DW and these parameters were less sensitive to the treatment than those in Williams in which the inhibition rate was 90% per plant and 80% per nodule g DW. These results indicate that NOD1-3 is partially nitrate-tolerant in terms of individual nodule growth as well as total nodule dry weight and N-2 fixation activity. A whole shoot of Williams and NOD1-3 plants was exposed to Co-14(2) for 120 min followed by 0 or 5 mm nitrate treatment for 2 d, and the partitioning of the photoassimilates among the organs was analyzed. Under 0 mm nitrate treatment, the percentages of the distribution of 14 C radioactivity between the nodules and roots were 63 and 37% in Williams and 89 and 11% in NOD1-3. Under the 5 mm nitrate conditions, the percentages of the distribution of C-14 between the nodules and roots changed to 14 and 86% in Williams and 39 and 61% in NOD1-3, respectively. These results indicated that the hypernodulating mutant NOD1-3 supplied a larger amount of photoassimilates to the nodules than to the roots under nitrogen-free conditions, and that the nitrate depression of photoassimilate transport to the nodules was less sensitive than that of the parent line.

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  • Site of nitrogen accumulation in tulip (Tulipa gesneriana L.) roots during winter 査読

    S Komiyama, S Tanabe, A Murayama, S Ruamrungsri, T Ikarashi, N Ohtake, K Sueyoshi, T Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   49 ( 4 )   611 - 618   2003年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    It was found that tulip (Tulipa gesneriana L.) plants accumulate a large amount of nitrogen in their roots during the winter season, which is efficiently used for rapid shoot growth after sprouting in spring. In the present paper, the site and origin of nitrogen accumulation in tulip roots were investigated in a vertical split-root experiment. Tulip roots were separated into upper and lower half parts in a two-compartment tray, and the culture solution was separately supplied for a week. Culture solution with N-free medium or that containing N-15-labeled nitrate (2 mm (NO3-)-N-15 + 0.5 mm NH4+) or N-15-labeled ammonium (2 mm NO3- + 0.5 mm (NH4+)-N-15) was supplied to individual trays to determine the metabolic fate of nitrate and ammonium nitrogen in either part of the roots. The results indicated that nitrate and ammonium absorption activities were similar between the lower and upper half-parts of the tulip roots. The absorbed N-15 predominantly accumulated either in the upper or lower part of the roots in direct contact with N-15-labeled medium, and only a small portion of N-15 was translocated to the other part. In the root part treated with N-15-labeled medium, most of the absorbed nitrogen was in ethanol soluble forms, and free glutamine and glutamic acid were intensely labeled with N-15. Also in the root parts directly in contact with nitrogen, the glutamine concentration in either the upper or the lower roots was considerably enhanced. On the other hand, nitrogen application did not affect the 4-methyleneglutamine concentration. Sucrose and glucose were the major soluble sugars in tulip roots, and the concentration was reduced at the nitrogen absorption site, possibly due to energy and carbon consumption for the absorption and assimilation of nitrogen.

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  • Quick and reversible inhibition of soybean root nodule growth by nitrate involves a decrease in sucrose supply to nodules 査読

    H Fujikake, A Yamazaki, N Ohtake, K Sueyoshi, S Matsuhashi, T Ito, C Mizuniwa, T Kume, S Hashimoto, NS Ishioka, S Watanabe, A Osa, T Sekine, H Uchida, A Tsuji, T Ohyama

    JOURNAL OF EXPERIMENTAL BOTANY   54 ( 386 )   1379 - 1388   2003年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    The upper part of a nodulated soybean root hydroponically cultured in a glass bottle was monitored using a computer microscope under controlled environmental conditions, and the diameter of individual nodules was measured from 10-24 d after planting. The diameter of a root nodule attached to the primary root increased from 1 mm to 6 mm for 2 weeks under nitrogen-free conditions. The increase in diameter of the nodules was almost completely stopped after 1 d of supplying 5 mM nitrate, and was due to the cessation of nodule cell expansion. However, nodule growth quickly returned to the normal growth rate following withdrawal of nitrate from the solution. The reversible depression of nodule growth by nitrate was similar to the restriction of photoassimilate supply by continuous dark treatment for 2 d followed by normal light/dark conditions. In addition, the inhibitory effect of nitrate was partially alleviated by the addition of 3% (w/v) sucrose to the medium. Plant leaves were exposed to C-11 or C-14-labelled carbon dioxide to investigate the effects of 5 mM nitrate on the translocation and distribution of photosynthates to nodules and roots. Supplying 5 mM nitrate stimulated the translocation rate and the distribution of labelled C in nitrate-fed parts of the roots. However, the C-14 partitioning to nodules decreased from 9% to 4% of total C-14 under conditions of 5 mM nitrate supply. These results indicate that the decrease in photoassimilate supply to nodules may be involved in the quick and reversible nitrate inhibition of soybean nodule growth.

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  • Effect of deep placement of calcium cyanamide, coated urea, and urea on soybean (Glycine max (L.) Merr.) seed yield in relation to different inoculation methods 査読

    T Kaushal, T Suganuma, H Fujikake, N Ohtake, K Sueyoshi, Y Takahashi, T Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   48 ( 6 )   855 - 863   2002年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    The main objective of this study was to increase the productivity of soybean [Glycine max (L.) Merr. cv, Enrei] seed by deep placement of controlled release nitrogen fertilizers and by the application of different., methods of inoculation of bradyrhizobia. Ten days old seedlings in an inoculated paper.,pot (IPP), in a non-inoculated paper pot (NIPP), and those grown in a vermiculite bed without paper pot (DT) were transplanted to an upland field converted from a drained paddy field in Nagaoka. In addition to the basal application of 16 kg N ha(-1) in the surface layer (Control), deep placement of 100 kg N ha-1 of urea (Urea), 100-day type coated urea (CU-100), and calcium cyanamide (CaCN2,) treatments were applied at the depth of 20 cm. In the IPP method, a significantly higher seed yield was obtained with the deep placement of CaCN2 and CU-100 compared with the Urea and Control treatments. A similar tendency was observed for the DT and NIPP methods. Among the same N fertilizer treatments, the seed yield for IPP and DT tended to exceed that for NIPP, although the NIPP roots also showed nodulation probably due to infection with indigenous bradyrhizobia. The percentage of nitrogen derived from atmospheric N, estimated by the simple relative ureide method was higher in the plants with CU-100 and CaCN2 compared with those with the Urea and Control treatments at the R1 stage, suggesting that the basal deep placement of CaCN2 or CU-100 for soybean cultivation enabled the supply of N without concomitant depression of N-2 fixation. Thus the deep placement of cheaper CaCN2 was found to be as effective as that of CU-100 for enhancing the soybean seed yield.

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  • A protein kinase activated by darkness phosphorylates nitrate reductase in Komatsuna (Brassica campestris) leaves 査読

    K Nakamura, N Shiraishi, S Hosoo, K Sueyoshi, T Sugimoto, T Nanmori, H Nakagawa, Y Oji

    PHYSIOLOGIA PLANTARUM   115 ( 4 )   496 - 503   2002年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL MUNKSGAARD  

    Although it has been shown that leaf nitrate reductase (NR: EC 1.6.6.1) is phosphorylated by subjecting plants to darkness, there is no evidence for the existence of dark-activated or dark-induced NR kinase. This study was undertaken to investigate the occurrence of a protein kinase phosphorylating NR in response to dark treatments. Immediately after transferring Komatsuna (Brassica campestris L.) plants to darkness, we observed rapid increases in the phosphorylating activity of the synthetic peptide, which is designed for the amino acid sequence surrounding the regulatory serine residue of the hinge 1 region of Komatsuna NR, in crude extracts from leaves. The activity reached a maximum after 10 min of darkness. Inactivation states of NR estimated from relative activities with or without Mg2+ were correlated to activities of the putative dark-activated protein kinase. Using the synthetic peptide as a substrate, we purified a protein kinase from dark-treated leaves by means of successive chromatographies on Q-Sepharose, Blue Sepharose, FPLC Q-Sepharose, and ATP-gamma-Sepharose columns. The purified kinase had an apparent molecular mass of 150 kDa with a catalytic subunit of 55 kDa, and it was Ca2+-independent. The purified kinase phosphorylated a recombinant cytochrome c reductase protein, a partial protein of NR, and holo NR, and inactivated NR in the presence of both 14-3-3 protein and Mg2+. The kinase also phosphorylated synthetic peptide substrates designed for sucrose phosphate synthase and 3-hydroxy-3-methylglutaryl-Coenzyme A reductase. Among inhibitors tested, only K252a, a potent and specific serine/threonine kinase inhibitor, completely inhibited the activity of the dark-activated kinase. The activity of the purified kinase was also specifically inhibited by K252a. Taken together with these findings, results obtained suggest that the putative dark-activated protein kinase may be the purified kinase itself, and may be responsible for in vivo phosphorylation of NR and its inactivation during darkness.

    DOI: 10.1034/j.1399-3054.2002.1150403.x

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  • Effect of exogenous salicylic acid supply on nodule formation of hypernodulating mutant and wild type of soybean

    Takashi Sato, Hiroyuki Fujikake, Norikuni Ohtake, Kuni Sueyoshi, Tadashi Takahashi, Atsushi Sato, Takuji Ohyama

    Soil Science and Plant Nutrition   48 ( 3 )   413 - 420   2002年6月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Informa UK Limited  

    DOI: 10.1080/00380768.2002.10409219

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  • Rapid and reversible nitrate inhibition of nodule growth and N-2 fixation activity in soybean (Glycine max (L.) Merr.) 査読

    H Fujikake, H Yashima, T Sato, N Ohtake, K Sueyoshi, T Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   48 ( 2 )   211 - 217   2002年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    Nodulated soybean (Glycine max. (L) Merr. cv. Williams) plants were hydroponically cultured, and various combinations of I-week culture with 5 or 0 mm nitrate were applied using 13-d-old soybean seedlings during three successive weeks. The treatments were designated as 0-0-0, 5-5-5, 5-5-0, 5-0-0, 5-0-5, 0-5-5, and 0-0-5, where the three sequential numbers denote the nitrate concentration (mm) applied in the first-second-third weeks. The size of the individual nodule was measured periodically using a slide caliper. All the plants were harvested after measurement of the acetylene reduction activity (ARA) at the end of the treatments. In the 0-0-0 treatment, the nodules grew continuously during the treatment period. Individual nodule growth was immediately suppressed after 5 mm nitrate supply. However, the nodule growth rapidly recovered by changing the 5 mm nitrate solution to a 0 mm nitrate solution in the 5-0-0 and 5-5-0 treatments. In the 5-0-5 treatment, nodule growth was completely inhibited in the first and the third weeks with 5 mm nitrate, but the nodule growth was enhanced in the second week with 0 mm nitrate. The nodule growth response to 5 mm nitrate was similar between small and large size nodules. After the 5-5-5, 5-0-5, 0-0-5, and 0-5-5 treatments, where the plants were cultured with 5 mm nitrate in the last third week, the ARA per plant was significantly lower compared with the 0-0-0 treatment. On the other hand, the ARA after the 5-0-0 and 5-5-0 treatments was relatively higher than that after the 0-0-0 treatment, possibly due to the higher photosynthate supply associated with the vigorous vegetative growth of the plants supplemented with nitrate nitrogen. It is concluded that both soybean nodule growth and N-2 fixation activity sensitively responded to the external nitrate level, and that these parameters were reversibly regulated by the current status of nitrate in the culture solution, possibly through sensing of the nitrate concentration in roots and / or nodules.

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  • Appearance of endopeptidases during the senescence of cucumber leaves 査読

    Y Yamauchi, T Sugimoto, K Sueyoshi, Y Oji, K Tanaka

    PLANT SCIENCE   162 ( 4 )   615 - 619   2002年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCI IRELAND LTD  

    In cucumber (Cucumis sativus L.) leaves at different ontogenic stages, a differential appearance of three major endopeptidases was observed by employing activity staining using gelatin as a substrate. On the basis of this observation, we discussed their physiological roles in senescing leaves. The most active endopeptidase in young mature leaves was a glutamyl endopeptidase with a pI of 4.5. It might be involved in active protein catabolism in young leaves because its activity became maximal just after the leaf had fully expanded and when protein and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) contents rapidly decreased. An endopeptidase with a pI of 4.3 was not observed in young leaves, however, it was highly active in senescing leaves. Interestingly, its activity in cotyledons was eliminated when the upper metabolically active leaves were removed. This implies that the appearance of this enzyme is regulated by the presence of sink tissues, and it is involved in the degradation of protein in senescing leaves facilitating N transfer to upper developing leaves. A trypsin-like endopeptidase with a pI of 5.0 showed relatively constant activity during the whole period. This endopeptidase has been shown to be inhibited by arginine, guanidino compounds and Mg2+, therefore, it might exist constitutively and its activity might be regulated mainly at a post-translational level responding to nutrient and environmental conditions. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.

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  • Effects of anion channel blockers on xylem nitrate transport in barley seedlings 査読

    T Kawachi, C Nishijo, H Fujikake, S Abdel-Latif, N Ohtake, K Sueyoshi, T Ohyama, N Shigeta-Ishioka, S Watanabe, A Osa, T Sekine, S Matsuhashi, T Ito, C Mizuniwa, T Kume, S Hashimoto, H Uchida, A Tsuji

    SOIL SCIENCE AND PLANT NUTRITION   48 ( 2 )   271 - 277   2002年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    The effects of anion channel blockers, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) and anthracene-9-carboxylic acid (A-9-C), on the uptake and xylem transport of nitrate were investigated in intact barley (Hordeum vulgare L.) seedlings using (NO3-)-N-13 and (NO3-)-N-15 as tracers. The seedling roots were pretreated with either blocker for 1 h and then (NO3-)-N-13 or (NO3-)-N-15 was supplied to the medium. Real-time images of N-13 accumulation in shoots were monitored during the first 30 min using Positron Emitting Tracer Imaging Systems (PETIS). The radioactivity in the shoots of the plants treated with either blocker was about half of that in the shoots of the control plants. Analysis of the distribution of N-13 in whole seedlings using a Bioimaging Analyzer System (BAS) showed a N-13-accumulation in both shoots and roots of the control plants but reduced N-13-levels in the shoots of the plants treated with either blocker. Nitrate concentrations in the xylem sap were significantly reduced by the application of either blocker while the net uptake of (NO3-)-N-15 was not or slightly influenced by the treatment with blockers. The translocation and uptake of chloride were also significantly reduced by the treatment with A-9-C but not with DIDS. These results suggested that anion channels contributed to xylem loading of nitrate in barley plants.

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  • Effect of short-term application of nitrogen on the accumulation of beta-subunit of beta-conglycinin in nitrogen-starved soybean (Glycine max L.) developing seeds 査読

    N Ohtake, T Kawachi, Okuyama, I, H Fujikake, K Sueyoshi, T Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   48 ( 1 )   31 - 41   2002年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    It has been reported that the accumulation of the beta-subunit of beta-conglycinin, a storage protein of soybean (Glycine max L.) seeds, was suppressed by nitrogen (N)-deficiency. In this report we attempted to determine which compound(s) may play a key role in regulating the accumulation of beta-subunit mRNA and protein in intact seeds and in an in vitro cotyledon culture system. Non-nodulating soybean plants were cultivated under N-deficient (0.5 mm NO3-) conditions, and transferred to N-sufficient medium (5 mm NO3-) when the plants reached the pod filling stage. The beta-subunit mRNA and the protein were detected in immature seeds within 2 d after transfer to 5 mm NO3- medium. Among the free amino acids in the immature seeds, asparagine concentration increased rapidly within 2 d. In the in vitro cotyledon culture system, the application of glutamine induced the accumulation of beta-subunit mRNA within 12 h, while asparagine did not induce the accumulation of beta-subunit mRNA for 7 d. An inhibitor of transaminases, (aminooxy)acetic acid, enhanced beta-subunit mRNA accumulation without Gln accumulation when asparagine was the sole nitrogen source. Although the asparagine concentration in the seed reflected the nitrogen statues in whole plant, the accumulation of beta-subunit mRNA and continuous protein storage in immature seeds appeared to be regulated by glutamine, its metabolites or related compounds.

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  • Role of xylem sap nitrate in the regulation of nitrate reductase gene expression in leaves of barley (Hordeum vulgare L.) seedlings 査読

    T Kawachi, Y Shoji, T Sugimoto, YL Oji, A Kleinhofs, RL Warner, N Ohtake, T Ohyama, K Sueyoshi

    SOIL SCIENCE AND PLANT NUTRITION   48 ( 1 )   79 - 85   2002年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    The role of xylem sap nitrate in the regulation of nitrate reductase (NR) in leaves was investigated in barley (Hordeum vulgare L.) seedlings. Upon the exposure of barley roots to nitrate at concentrations ranging from 0.1 to 5 mm, rapid accumulation of NADH-specific NR mRNA was observed in both leaves and roots. In the same treatment, the concentration of xylem sap nitrate also rapidly increased during the early feeding period and reached a steady state at concentration ranging from 15 to 50 mm. Patterns of NR mRNA accumulation in leaves were closely linked with the increase in the nitrate concentration in the xylem sap, but not with the amount of nitrate stored in the leaf tissues. High levels of NR mRNA in intact leaves were observed only when the xylem sap nitrate reached mm levels. Supply of different concentrations of nitrate to detached leaves also indicated that mM levels of nitrate supplied through the xylem were required for adequate accumulation of NR mRNA in leaves. When nitrate-grown seedlings were transferred to a nitrate-free medium, a parallel decline in both the concentration of xylem sap nitrate and NR mRNA level was observed, while the nitrate concentrations in leaf tissues remained constant. These results indicate that continuous supply of mM levels of nitrate through the xylem is important for maintaining the high levels of NR mRNA in barley leaves.

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  • Expression of asparagine synthetase in rice (Oryza sativa) roots in response to nitrogen 査読

    T Kawachi, K Sueyoshi, A Nakajima, H Yamagata, T Sugimoto, Y Oji

    PHYSIOLOGIA PLANTARUM   114 ( 1 )   41 - 46   2002年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL MUNKSGAARD  

    The expression of asparagine synthetase (AS; EC 6.3.5.4) in response to externally supplied nitrogen was investigated with respect to enzyme activity and protein levels as detected immunologically in rice (Oryza sativa) seedlings. The asparagine content was very low in leaves and roots of nitrogen-starved rice plants but increased significantly after the supply of 1 mM NH4+ to the nutrient solution. While neither AS activity nor AS protein could be detected in leaves and roots prior to the supply of nitrogen, levels became detectable in roots but not in leaves within 12 h of the supply of I mM NH4+ Or 10 mM glutamine. Other nitrogen compounds, such as nitrate, glutamate, aspartate and asparagine had no effect. Methionine sulfoximine completely inhibited the NH4+-induced accumulation of AS protein but did not affect the glutamine-induced accumulation of the enzyme. The results suggested that glutamine or glutamine-derived metabolites regulate AS expression in rice roots.

    DOI: 10.1034/j.1399-3054.2002.1140107.x

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  • A serine endopeptidase from cucumber leaves is inhibited by L-arginine, guanidino compounds and divalent cations 査読

    Y Yamauchi, T Sugimoto, K Sueyoshi, Y Oji, K Tanaka

    PHYTOCHEMISTRY   58 ( 5 )   677 - 682   2001年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    An endopeptidase was purified and characterized from green leaves of cucumber (Cucumis sativus L. suyo). The purified enzyme, a basic amino acid-specific endopeptidase with a pI of 5.0, was a monomeric protein of 80 kDa whose pH optimum was 9.5. Inhibitor analysis suggested that it was a serine endopeptidase and contained sulfhydryl groups essential for catalytic activity. Analysis of internal amino acid sequences of the endopeptidase showed no significant similarity to other proteins. Its activity was inhibited by L-Arg and guanidino compounds having high hydrophobicity, as well as divalent cations such as Mg2+ and Ca2+. The K-i values of L-Arg and Mg2+, which are also likely in vivo inhibitors, were 3.5 and 10 mM, respectively. Inhibition by L-Arg and Mg2+ was additive, and more than 70% of the activity was reversibly inhibited under their physiologically significant concentrations. These results suggest that the enzyme is possibly regulated by L-Arg and/or guanidino compounds, and by divalent cations in vivo. (C) 2001 Elsevier Science Ltd. All rights reserved.

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  • A high molecular weight glutamyl endopeptidase and its endogenous inhibitors from cucumber leaves

    Y Yamauchi, Y Ejiri, T Sugimoto, K Sueyoshi, Y Oji, K Tanaka

    JOURNAL OF BIOCHEMISTRY   130 ( 2 )   257 - 261   2001年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE BIOCHEMICAL SOC  

    We purified a glutamyl endopeptidase that is a major foliar endopeptidase in cucumber. The endopeptidase had a molecular mass of 400 kDa, consisted of four subunits of 97 kDa, and was inactivated by SH-modifying reagents. Its optimum pH and optimum temperature were 8.0 and 30-37 degreesC, respectively. An internal amino acid sequence of the endopeptidase was highly homologous to a partial sequence of unidentified proteins deduced from genetic information for Arabidopsis thaliana, soybean and rice, but not to the sequences of bacterial glutamyl endopeptidases or animal proteases. Therefore, the unidentified proteins might be glutamyl endopeptidases and be widely distributed only among plant species. The activity of the cucumber glutamyl endopeptidase was inhibited by at least three inhibitors existing in cucumber leaves. One of the inhibitors was a competitive inhibitor of 25 kDa, which did not significantly inhibit commercial endopeptidases derived from animals and microorganisms. This suggests that the cucumber glutamyl endopeptidase might be controlled by endogenous inhibitors in vivo.

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  • Temporary application of nitrate to nitrogen-deficient soybean plants at the mid- to late-stages of seed development increased the accumulation of the beta-subunit of beta-conglycinin, a major seed storage protein 査読

    N Ohtake, T Kawachi, A Sato, Okuyama, I, H Fujikake, K Sueyoshi, T Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   47 ( 1 )   195 - 203   2001年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    The accumulation of the beta -subunit of beta -conglycinin, a major soybean storage protein, in seeds is controlled by the nitrogen (N) status of the plant. When non-nodulating soybean mutants (T201) were hydroponically cultivated under N-sufficient conditions (supply of 5 mM NaNO3 to the medium), beta -subunit accumulation was first detected at 30 d after flowering (DAF). In N-deficient plants (supply of 1 mM NaNO3), the accumulation of the beta -subunit was strongly inhibited. When 5 mM N-15-labeled NO3- was supplied for 1 week to N-deficient T201 plants before 28 DAF, the beta -subunit protein did not accumulate, but the application of 5 mM NO3- after 28 DAF, led to a substantial accumulation of the beta -subunit, The total N-15 percentage in mature seeds was similar, irrespective of the time of feeding, regardless of the accumulation of the beta -subunit. The 7 S (mostly composed of beta -conglycinin) and 11 S globulins (mostly glycinin, which was another major component of storage protein) contained N from 5 mM (NO3-)-N-15 at a similar level, about 60% of total N, These results indicate that a) the temporary supply of a high level of NO3- effectively promoted beta -subunit accumulation in N-deficient plants when feeding was carried out after 28 DAF, and b) incorporation of fed NO3- was similar between 7 S and 11 S globulins.

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  • Rapid N transport to pods and seeds in N-deficient soybean plants 査読

    N Ohtake, T Sato, H Fujikake, K Sueyoshi, T Ohyama, NS Ishioka, S Watanabe, A Osa, T Sekine, S Matsuhashi, T Ito, C Mizuniwa, T Kume, S Hashimoto, H Uchida, A Tsuji

    JOURNAL OF EXPERIMENTAL BOTANY   52 ( 355 )   277 - 283   2001年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    Non-nodulated soybean (Glycine max (L,) Merr,) plants were cultivated hydroponically under N-sufficient (5 mM NaNO3) or N-deficient (0.5 mM NaNO3) conditions. N-13- Or N-15-labelled nitrate was fed to the cut end of the stems, and the accumulation of nitrate-derived N in the pods, nodes and stems was compared. Real-time images of N-13 distribution in stems, petioles and pods were obtained using a Positron Emitting Tracer Imaging System for a period of 40 min. The results indicated that the radioactivity in the pods of N-deficient plants was about 10 times higher than that of N-sufficient plants, although radioactivity in the stems and nodes of N-deficient versus N-sufficient plants was not different. A similar result was obtained by supplying (NO3-)-N-15 to cut soybean shoots for 1 h, The fact that the N translocation into the pods from NO3- fed to the stem base was much faster in N-deficient plants may be due to the strong sink activity of the pods in N-deficient plants. Alternatively, the redistribution of N from the leaves to the pods via the phloem may be accelerated in N-deficient plants. The temporal accumulation of (NO3-)-N-13 in nodes was suggested in both N-sufficient and N-deficient plants. In one (NO3-)-N-13 pulse-chase experiment, radioactivity in the stem declined rapidly after transferring the shoot from the (NO3-)-N-13 solution to non-labelled NO3; in contrast, the radioactivity in the node declined minimally during the same time period.

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  • Changes in four leghemoglobin components in nodules of hypernodulating soybean (glycine max [L] merr.) mutant and its parent in the early nodule developmental stage 査読

    Takashi Sato, Noriyasu Onoma, Hiroyuki Fujikake, Norikuni Ohtake, Kuni Sueyoshi, Takuji Ohyama

    Plant and Soil   237 ( 1 )   129 - 135   2001年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Soybean nodules contain four major leghemoglobin (Lb) components, Lba, Lbc1, Lbc2 and Lbc3. A sensitive and selective method for quantitative analysis of the four Lb components was devised with capillary isoelectric focusing (CIEF). The changes in the concentrations of four Lb components in nodules during the initial stages of development were compared between hypernodulating soybean mutant NOD1-3 and its parent cv. Williams. The hydroponically cultivated soybean plants were periodically sampled. All the visible nodules were collected from the roots, and then the four Lb components in the largest nodules were analyzed with the CIEF method. In NOD1-3 Lbs were initially detected at 19 days after sowing (DAS), a few days earlier than in Williams at 22 DAS. The Lbcs (Lbc1, Lbc2 and Lbc3) were the main component at the earliest nodule growth stage, and the relative proportion of Lba increased with nodule growth in both NOD1-3 and Williams. This result is in agreement with previous observation, and the CIEF method is considered to be useful for Lb components analysis to define their function and gene expression.

    DOI: 10.1023/A:1013317219871

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  • Effects of inhibitors for signaling components on the expression of the genes for nitrate reductase and nitrite reductase in excised barley leaves

    K Sueyoshi, T Mitsuyama, T Sugimoto, A Kleinhofs, RL Warner, Y Oji

    SOIL SCIENCE AND PLANT NUTRITION   45 ( 4 )   1015 - 1019   1999年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    Nitrate-induced accumulation of mRNAs for nitrate reductase and nitrite reductase was strongly inhibited by treatment with inhibitors of Ca2+ channels, protein phosphatases, and tyrosine protein kinases in excised barley leaves. It was shown that these components may be involved in the nitrate-response signaling pathways for the expression of nitrate and nitrite reductase genes in barley leaves.

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  • Ethylene-induced putrescine accumulation modulates K+ partitioning between roots and shoots in barley seedlings 査読

    T Tamai, M Inoue, T Sugimoto, K Sueyoshi, N Shiraishi, Y Oji

    PHYSIOLOGIA PLANTARUM   106 ( 3 )   296 - 301   1999年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MUNKSGAARD INT PUBL LTD  

    We investigated the cause and effect relationships among ethylene, polyamines, and K+ in barley (Hordeum vulgare L. cv. Amagi) seedlings, Application of 1-aminocyclopropane-1-carboxylic acid (ACC), a precursor of ethylene, to the growth medium caused a decrease in K+ concentration in roots and an increase in shoots. Addition of ACC induced putrescine accumulation in roots, while spermidine and spermine levels remained unchanged. Exogenous supply of putrescine led to putrescine accumulation and reduced K+ concentration. Application of Co2+, an inhibitor of ethylene biosynthesis, together with ACC, inhibited putrescine accumulation with a decrease in K+ concentration in roots, ACC-treated roots showed K+ uptake capacity equivalent to that of control roots, implying that the majority of K+ is translocated to shoots. These results suggest that ethylene regulates K+ partitioning between roots and shoots through the level of accumulation of putrescine in barley seedlings.

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  • Possible involvement of photosynthetic supply in changes of nodule characteristics of hypernodulating soybeans 査読

    T Sato, H Yashima, N Ohtake, K Sueyoshi, S Akao, T Ohyama

    SOIL SCIENCE AND PLANT NUTRITION   45 ( 1 )   187 - 196   1999年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    The hypernodulating mutant of soybean En6500 has a larger number of small nodules than its parent, cv. Enrei, and the shoot size is relatively small at the same time. It was shown that some characteristics of the nodules of En6500 were different from those of the parent nodules such as, nodule size distribution, acetylene reduction activity (ARA), concentration of leghemoglobin (Lb) components and proportion of the infected region in the nodules. In this study, we attempted to identify the main factor(s) which control(s) the changes in the characteristics of mutant nodules, by manipulating the source-sink relationship in relation to the photosynthetic supply.
    By decreasing the infection dose of Bradyrhizobium japonicum, the nodule number of En6500 could be reduced to the level of nodulation normally observed in Enrei. In addition a part of Enrei shoot was cut off for equalizing the shoot size of Enrei with that of En6500 at 16 d after sowing (DAS). The plants were hydroponically cultured until 30 DAS with or without 5 mM NO3- for the last 14 d. ARA, bacteroid density, concentration of Lb components (Lba, Lbc1, Lbc2, and Lbc3), and volume ratio of the infected region in the nodules were determined.
    When the hypernodulating mutant En6500 which was treated to obtain a similar nodule number to that of Enrei was compared with the parent Enrei which was treated to obtain the same shoot size as that of En6500, nodule characteristics such as, ARA per nodule dry weight, Lb concentration, Lb component ratios and volume ratio of the infected region in the nodules, were found to be similar in both En6500 and Enrei. These results suggest that the specific characteristics of the nodules of hypernodulating mutant, such as low ARA, low Lb concentration and small infected region, may be caused by the insufficient supply of photosynthates to each nodule. It is concluded that many of the specific characteristics described above in the nodules of hypernodulating En6500 are not directly related to genetic factors induced by chemical mutation, but that a secondary effect is involved in that the mutants have an excess number of nodules with a relatively small shoot. On the other hand, the partial tolerance of nodule growth and ARA per plant to nitrate may be involved in another mechanism than the secondary effect of photosynthate deficiency.
    In soybean, autoregulatory control may play an important role in optimizing the nodule number where the nodule growth and total N-2 fixation activity can be maximized.

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  • 70 窒素供給時期がダイズ種子貯蔵タンパク質組成に及ぼす影響(関東支部講演会)

    大竹 憲邦, 佐藤 明世, 長谷 光展, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   45   402 - 402   1999年

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

    DOI: 10.20710/dohikouen.45.0_402_2

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  • 10-16 窒素栄養がダイズ種子内のIAAに及ぼす影響(10.植物の代謝)

    佐藤 明世, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   45   124 - 124   1999年

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

    DOI: 10.20710/dohikouen.45.0_124_2

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  • 10-13 窒素によるダイズ種子貯蔵タンパク質の集積調節機構の解析(10.植物の代謝)

    大竹 憲邦, 佐藤 明世, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   45   123 - 123   1999年

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

    DOI: 10.20710/dohikouen.45.0_123_1

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  • 68 14-3-3タンパク質による植物硝酸還元酵素の活性調節(関東支部講演会)

    林 優子, 今井 光, 末吉 邦, 大竹 憲邦, 王子 善清, 大山 卓爾

    日本土壌肥料学会講演要旨集   45   401 - 401   1999年

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

    DOI: 10.20710/dohikouen.45.0_401_2

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  • Purification of inactivation factor of phosphorylated Nitrate reductase from Leaves of Brassica campestris 査読

    K Sueyoshi, M Kojima, T Sugimoto, H Nakagawa, Y Oji

    JOURNAL OF PLANT PHYSIOLOGY   153 ( 1-2 )   74 - 78   1998年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:GUSTAV FISCHER VERLAG  

    The factor that inactivates the phosphorylated nitrate reductase was purified 870-fold from leaves of Brassica campestris L. ssp. apifera by column chromatography oil DEAE-Toyopearl, Phenyl Sepharose, Sephadex G-200 and Mono Q. Final preparation of the purified factor was nearly homogenous as revealed by polyacrylamide gel electrophoresis and its molecular weight was estimated to be 70 ku using Superose 12 gel filtration column. SDS-polyacrylamide gel analysis of the final preparation showed two protein bands with molecular weight of 30 and 33 ku suggesting that the inactivation factor was composed of two heterogenous subunits. The inactivation of phosphorylated nitrate reductase by the purified protein was neither rime dependent, nor due to degradation of nitrate reductase, suggesting chat the inactivation factor exhibits its effect by binding to the phosphorylated nitrate reductase.

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  • Determination of leghemoglobin components and xylem sap composition by capillary electrophoresis in hypernodulation soybean mutants cultivated in the field 査読

    Takashi Sato, Hiroyuki Yashima, Norikuni Ohtake, Kuni Sueyoshi, Shoichiro Akao, James E. Harper, Takuji Ohyama

    Soil Science and Plant Nutrition   44 ( 4 )   635 - 645   1998年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The hypernodulation soybean mutant lines (NOD1-3, NOD2-4, NOD3-7) and their parent Williams, and the mutant En6500 and its parent Enrei were cultivated in a sandy dune field in Niigata, and the nodules and root bleeding xylem sap were sampled at 50, 70, 90, and 120 d after planting (DAP). The nodule size distribution patterns and concentration of leghemoglobin components were determined. The number of nodules of the hypernodulation mutant lines was about two to three times higher than that of the parent lines irrespective of the sampling date. At 50 DAP the nodule size was relatively smaller in the hypernodulation mutant lines, and the total dry weight of the nodules was almost the same in the mutant lines and their parents. At 70 DAP and 90 DAP, the size distribution of the hypernodulation mutant nodules became.almost the same as that of the parent lines, and both the number and total dry weight of the nodules were higher than those of the parent lines. The concentration of four Lb components was separately measured by capillary electrophoresis. The concentration of the Lb components in the hypernodulation mutant lines tended to be lower than in the parents, but the component ratios were not different between the hypernodulation mutants and their parents. Under field conditions, plant growth and nodulation characteristics were more similar between mutants and parents than in the hydroponic culture reported previously, although the mutants did exhibit hypernodulation traits. These findings suggest that the decrease in the Lb concentration and the different Lb components ratios in the mutants may be caused by secondary effects of excess nodulation, such as photosynthate deficiency, rather than by a genetic defect in mutation. The concentration of major nitrogenous compounds (allantoic acid, allantoin, asparagine, aspartic acid, and nitrate) in the xylem sap was also measured by capillary electrophoresis. The concentration of ureides and nitrate in xylem sap decreased with the plant age, but the asparagine concentration increased during the same period. The concentrations of ureides and asparagine were higher, and the nitrate concentration was lower in the mutant lines than in their parents, possibly due to the higher dependence on N2 fixation than N03 ™ utilization. In the xylem sap, nitrate was the major inorganic anion followed by phosphate, sulfate, and chloride, and potassium was the major cation followed by calcium or magnesium and sodium. © 1998, Taylor &amp
    Francis Group, LLC.

    DOI: 10.1080/00380768.1998.10414487

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  • Effect of nitrogen application at the flowering stage on the quality of soybean seeds 査読

    T Sugimoto, K Nomura, R Masuda, K Sueyoshi, Y Oji

    JOURNAL OF PLANT NUTRITION   21 ( 10 )   2065 - 2075   1998年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MARCEL DEKKER INC  

    Application of nitrogen (N) fertilizer at the flowering stage changed the contents of storage compounds in seeds of Soybean (Glycine max L. cvs Enrei and Tamahomare). The effects of the N application on the maturation of soybean seeds were examined by comparing changes in the contents of amino acids, sugars, water, protein, and oil in seeds from N-dressed plants (NDS) with those from undressed plants (UDS) during maturation. The application resulted in a decrease of contents of total and some amino acids (glutamine and asparagine) in developing seeds except for at the early maturation stage and in a decreased protein content of mature seeds. On the other hand, the N application led to faster accumulation of oil in developing seeds and to an increased oil content of mature seeds. Based on these results, it was concluded that the N application at the flowering stage changed the composition of solutes imported by developing seeds and resulted in variations in the contents of storage compounds. The results indicate that it is possible to improve seed quality by fine control of N application.

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  • Characterization of plasma membrane-bound Fe3+-chelate reductase from Fe-deficient and Fe-sufficient cucumber roots

    K Sueyoshi, O Hirata, Y Oji

    SOIL SCIENCE AND PLANT NUTRITION   43 ( 1 )   149 - 156   1997年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    In vivo Fe3+-chelate reductase (FeR) activity in cucumber roots (Cucumis sativus L,) increased by transferring the plants from Fe-sufficient (+Fe) conditions to Fe-deficient (-Fe) conditions, This increase was inhibited by the protein synthesis inhibitor, cycloheximide, The plasma membranes were isolated from the +Fe and -Fe roots and then the enzymatic properties of plasma membrane-bound FeR were characterized, The FeR in the plasma membranes from both the +Fe and -Fe roots reduced Fe3+-citrate using NADH as an electron donor in the presence of Triton X-100, Plasma membrane-bound FeR from both types of roots showed similar K-m values for Fe3+-citrate and NADH at 70 and 100 mu M, respectively, whereas V-max of the enzyme from -Fe roots was three-fold higher than that of the enzyme from +Fe roots, The enzyme was solubilized from plasma membranes with 1.0% Triton X-100 and subsequently analyzed on an isoelectric focusing gel, The activity staining of the gel after electrophoresis showed that four FeR isozymes with different pls of 5.3, 6.8, 7.5, and 8.7 were present on the plasma membranes of both the +Fe and -Fe roots, Only the intensity of the pi 7.5 band was enhanced in the -Fe roots, These results suggested that the increase of FeR activity in the -Fe roots resulted from the increased synthesis of the FeR isoform which is constitutively present in the plasma membranes of +Fe roots.

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  • 10-9 イネ幼植物アスパラギン合成酵素の窒素に対する応答(10.植物の代謝)

    川地 太兵, 末吉 邦, 杉本 敏男, 王子 善清

    日本土壌肥料学会講演要旨集   43   104 - 104   1997年

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

    DOI: 10.20710/dohikouen.43.0_104_1

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  • Emergence of proteases in germinating cucumber cotyledons and their roles in the two-step degradation of storage protein 査読

    Y Yamauchi, T Sugimoto, K Sueyoshi, Y Oji

    PLANT AND CELL PHYSIOLOGY   37 ( 3 )   279 - 284   1996年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    The degradation of storage protein in germinating cucumber seeds was shown to proceed via two distinct steps. First, several proteases with acidic isoelectric points (pIs) were involved in solubilization and partial degradation of 11S globulin, Treatment of seedlings with cycloheximide inhibited this step and the expression of these proteases. Thus, the first step appeared to be governed by these proteases, which were synthesized de novo after imbibition, The first step was observed in dark-grown cotyledons, but the complete degradation of 11S globulin did not occur in the absence of illumination, An additional protease, with a pI of 4.5, was induced by illumination, and it was involved in the further cleavage of the partially degraded products of 11S globulin. Thus, the complete degradation of the storage protein proceeded via a two-step process in illuminated germinating seedlings, Light is needed to induce the second step in the degradation of 11S globulin that supplies the nitrogen required for development of the photosynthetic apparatus in the greening cotyledon.

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  • EXPRESSION OF NADH-SPECIFIC AND NAD(P)H-BISPECIFIC NITRATE REDUCTASE GENES IN RESPONSE TO NITRATE IN BARLEY 査読

    K SUEYOSHI, A KLEINHOFS, RL WARNER

    PLANT PHYSIOLOGY   107 ( 4 )   1303 - 1311   1995年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC PLANT PHYSIOLOGISTS  

    Barley (Hordeum vulgare L.) has two, differentially regulated, nitrate reductase (NR) genes, one encoding the NADH-specific NR (Nar1) and the other encoding the NAD(P)H-bispecific NR (Nar7). Regulation of the two NR genes by nitrate was investigated in wild-type Steptoe and in an NADH-specific NR structural gene mutant (Az12). Gene-specific probes were used to estimate NADH and NAD(P)H NR mRNAs. The kinetics of induction by nitrate were similar for the two NR genes; expression was generally below the limits of detection prior to induction, reached maximum levels after 1 to 2 h of induction in roots and 4 to 8 h of induction in leaves, and then declined to steady-state levels. Derepression of the NAD(P)H NR gene in leaves of the NADH-specific NR gene mutant Az12 did not appear to be associated with changes in nitrate assimilation products or nitrate flux. Nitrate deprivation resulted in rapid decreases in NADH and NAD(P)H NR mRNAs in seedling roots and leaves and equally rapid decreases in the concentration of nitrate in the xylem sap. These results indicate that factors affecting nitrate uptake and transport could have a direct influence on NR expression in barley leaves.

    DOI: 10.1104/pp.107.4.1303

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  • イネ幼植物切断根のH^+/K^+交換輸送に対する原形質膜H-ATPase阻害剤,およびイオノフォアの効果

    原田 久富美, 藤本 健, 脇内 成昭, 末吉 邦, 王子 善清

    日本土壌肥料学雑誌   63 ( 1 )   39 - 45   1992年

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    記述言語:日本語   出版者・発行元:日本土壌肥料學會  

    イネ切断根による H^+/K^+ 交換輸送および再構成原形質膜小胞の ATPase 活性と pH 勾配形成能に対する ATPase 阻害剤およびイオノフォアの効果を調べた.適当な濃度の阻害剤およびイオノフォアは,K^+ 吸収と H^+ 排出の両方を完全に阻害したが,ATPase 活性および pH 勾配形形成能を阻害しなかった.したがって,H^+/K^+ 交換輸送は,原形質膜 ATPase により原形質膜を隔てて存在する H^+ の電気化学ポテンシャル差により制御されていると思われた.とくに,阻害剤やイオノフォアを添加することにより,H^+/K^+ 交換輸送は K^+ が放出される方向にも進行することが示された.また,K^+ 以外の1価カチオンを用いた場合,Rb^+, NH_4^+ では K^+ と同様に吸収溶液の pH の低下が生じたが,一方,Na^+, LI^+, Cs^+ の場合には,大きな pH 変化は認められなかった.

    DOI: 10.20710/dojo.63.1_39

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  • RECONSTITUTION AND CHARACTERIZATION OF PLASMA-MEMBRANE H+-ATPASE ACTIVITY FROM RICE ROOTS

    H HARADA, N WAKIUCHI, K SUEYOSHI, Y OJI

    SOIL SCIENCE AND PLANT NUTRITION   37 ( 4 )   725 - 733   1991年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC SOIL SCIENCE PLANT NUTRITION  

    Plasma membrane vesicles were purified from rice (Oryza sativa L. cv. Nipponbare) roots using aqueous two-phase partitioning. The purified vesicles were solubilized with deoxycholate and reconstituted with soybean phospholipids by gel filtration. Ionophores stimulated the ATPase activity of the reconstituted vesicles. Inhibition of the ATPase activity by vanadate increased from 77 to 93% by the reconstitution. These results indicate that the reconstituted vesicles were sealed and the vanadate-sensitive ATPase activity was purified during the reconstitution. The ATPase activity of the reconstituted vesicles was characterized. Only ATP was effectively hydrolyzed. Apparent K(m) value was 1.26 mm. Divalent cations were necessary for the activity, and their stimulation was in the following order: Mg2+ > Fe2+ > Mn2+, Co2+ >> Ca2+. Half-maximal inhibition by vanadate occurred at 5-mu-M. Monovalent ions stimulated the activity, and cations and anions were effective in the order of NH4+ > K+, Rb+ > Na+ > Li+ > choline+ and NO3- > Cl-, Br- > I- > SO42-, respectively. Optimum pH was 6.5.

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  • オオムギ根液胞膜のプロトン輸送性ピロフォスファターゼに及ぼす各種イオンの影響について

    仲尾次 浩一, 原田 久富美, 脇内 成昭, 末吉 邦, 王子 善清, 志賀 一一

    日本土壌肥料学雑誌   62 ( 4 )   393 - 398   1991年

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    記述言語:日本語   出版者・発行元:日本土壌肥料學會  

    オオムギ幼植物根からデキストランの密度勾配遠心分離法により,0%,と6%の界面に回収された膜小胞は,ATPおよびPP_i依存のプロトン輸送能をもっていた。ATPおよびPP_i依存性プロトン輸送はそれぞれ硝酸塩およびフッ化物イオンにより阻害されたことから,この調製した膜小胞は液胞膜由来であると考えられる。液胞膜小胞におけるPP_i分解活性およびPP_i依存性プロトン輸送活性は完全にカリウムに依存していた。それぞれの活性はルビジウムの存在下ではカリウムと同程度の活性が認められたが,ナトリウムとリチウムではカリウムの代替性はなかった。液胞膜小胞のPP_i分解活性はアニオンにより活性に差異が認められなかったが,PP_i依存性プロトン輸送活性はアニオンによって著しい影響を受け,活性の強さの順序はNO_3^->Br^->Cl^-≫SO_4^<2->の順であり,透過性アニオンの非存在下では活性はみられなかった。プロトン輸送はNO_3^-とCl^-の濃度に対して飽和曲線を示し,アニオンの輸送担体が存在することが推察された。

    DOI: 10.20710/dojo.62.4_393

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  • 6-31 キュウリ緑葉オルニチントランスカルバミラーゼの精製と性質(6. 植物の代謝および代謝成分)

    末吉 邦, 王子 善清

    日本土壌肥料学会講演要旨集   37   99 - 99   1991年

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

    DOI: 10.20710/dohikouen.37.0_99_1

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  • 5-32 イネ切断根におけるカリウムの吸収とそれに伴うプロトンの放出について(5. 植物の無機栄養および養分吸収)

    原田 久富美, 脇内 成昭, 藤本 健, 末吉 邦, 王子 善清

    日本土壌肥料学会講演要旨集   37   73 - 73   1991年

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

    DOI: 10.20710/dohikouen.37.0_73_2

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  • 25 オオムギ根液胞膜のプロトン輸送性ピロフォスファターゼについて(関西支部講演会講演要旨)

    仲尾次 浩一, 原田 久富美, 脇内 成昭, 末吉 邦, 王子 善清, 志賀 一一

    日本土壌肥料学会講演要旨集   37   340 - 340   1991年

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

    DOI: 10.20710/dohikouen.37.0_340_1

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  • IDENTIFICATION OF POSSIBLE INTERMEDIATES IN INVIVO DEGRADATION OF SPINACH NITRATE REDUCTASE

    K SUEYOSHI, N OGURA, H NAKAGAWA

    AGRICULTURAL AND BIOLOGICAL CHEMISTRY   53 ( 1 )   151 - 156   1989年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPAN SOC BIOSCI BIOTECHN AGROCHEM  

    Degradation intermediates of nitrate reductase (NR) were extracted from fresh spinach leaves in the presence of protease inhibitors, and concentrated by chromatography on an anti-NR IgG-conjugated Sepharose-4B column. Seven polypeptides, 120 (that of the intact subunit), 110, 100, 76, 64, 62, and 44 kDa, were obtained. Their cross-reactivities to mono-specific polyclonal antibody and monoclonal antibody prepared against NR subunit were assessed by an immunoblotting method. All polypeptides were recognized with mono-specific polyclonal antibody. On the other hand, four of them, at 120, 110, 100, and 44 kDa, were recognized with monoclonal antibody which specifically binds to the NADH-ferricyanide reductase domain of NR. The possibility that these polypeptides were artifacts in preparation was ruled out by an internal tracer experiment. Thus, it is concluded that spinach leaf NR <i>in vivo</i> degraded <i>via</i> putative intermediates catabolic products of 110, 100, 76, 64, 62, and 44 kDa.

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  • MONOCLONAL-ANTIBODIES SPECIFIC FOR NADH-FERRICYANIDE REDUCTASE DOMAIN OF SPINACH LEAF NITRATE REDUCTASE

    K SUEYOSHI, Y KUBO, K YAMAGISHI, N OGURA, K OCHIAI, K FUKUSHIMA, T IKEDA, H NAKAGAWA

    PLANT AND CELL PHYSIOLOGY   29 ( 6 )   975 - 980   1988年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    Two monoclonal antibodies, 17(3)9 and 36(79)4, were prepared against nitrate reductase from Spinacia oleracea L. leaves. An enzyme-linked immunosorbent assay showed that 17(3)9, but not 36(79)4, reacted more strongly to heat-denatured than native antigen. These antibodies inhibited NADH-nitrate reductase as well as its various partial activities including reduced methyl vilogen-nitrate reductase, reduced flavin mononucleotide-nitrate reductase and NADH-cytochrome c reductase activities, but not NADH-ferricyanide reductase activity. Immunoblotting after electrophoretic separation of nitrate reductase fragments obtained by Staphyrococcus aureus V8 protease digestion of native enzyme revealed that the two monoclonal antibodies bind to different epitopes located on the 28 kDa of the NADH-ferricyanide reductase domain.

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  • ACTIVITIES AND PROPERTIES OF PEPTIDYLARGININE DEIMINASES OF SEVERAL VERTEBRATE BRAINS

    H TAKAHARA, K SUEYOSHI, K SUGAWARA

    AGRICULTURAL AND BIOLOGICAL CHEMISTRY   50 ( 5 )   1303 - 1306   1986年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPAN SOC BIOSCI BIOTECHN AGROCHEM  

    Extracts from brains of mouse, rat, rabbit, pig, chicken, and duck were assayed for peptidylarginine deiminase, which catalyzes the deimination of arginyl residues. There was peptidylarginine deiminase in the extracts from all the brains that we tested, suggesting the widespread occurrence of this enzyme in vertebrate brains. The levels of the activity in the brains of 8 different inbred strains of mice could be divided into two groups. There is a difference of over 300% between the high and low groups. Furthermore peptidylarginine deiminase purified from the brains of mouse, pig, and chicken showed that there is no great difference in their MW and the optimal conditions for the activity. However, the substrate specificity of the enzyme from chicken brain was somewhat different from those of the enzymes from mammalian brains.

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  • Recent Advances in Carbon and Nitrogen Metabolism in C3 Plants 国際誌

    Marouane Baslam, Toshiaki Mitsui, Kuni Sueyoshi, Takuji Ohyama

    INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES   22 ( 1 )   2021年1月

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    記述言語:英語   出版者・発行元:MDPI  

    C and N are the most important essential elements constituting organic compounds in plants. The shoots and roots depend on each other by exchanging C and N through the xylem and phloem transport systems. Complex mechanisms regulate C and N metabolism to optimize plant growth, agricultural crop production, and maintenance of the agroecosystem. In this paper, we cover the recent advances in understanding C and N metabolism, regulation, and transport in plants, as well as their underlying molecular mechanisms. Special emphasis is given to the mechanisms of starch metabolism in plastids and the changes in responses to environmental stress that were previously overlooked, since these changes provide an essential store of C that fuels plant metabolism and growth. We present general insights into the system biology approaches that have expanded our understanding of core biological questions related to C and N metabolism. Finally, this review synthesizes recent advances in our understanding of the trade-off concept that links C and N status to the plant's response to microorganisms.

    DOI: 10.3390/ijms22010318

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  • ダイズの種子収量、元素集積、貯蔵タンパク組成に対するリン供給量の影響

    大竹 憲邦, 七夕 小百合, 五十嵐 弥重, 白鳥 豊, 本間 利光, 南雲 芳文, 高橋 能彦, 末吉 邦, 大山 卓爾, 大竹 憲邦, 七夕 小百合, 五十嵐 弥重, 白鳥 豊, 本間 利光, 南雲 芳文, 高橋 能彦, 末吉 邦, 大山 卓爾

    新潟大学農学部研究報告 = Bulletin of the Faculty of Agriculture, Niigata University   69   25 - 31   2017年2月

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    記述言語:英語   出版者・発行元:新潟大学農学部  

    Various levels of P application were imposed to hydroponically grown soybean plants, and the concentrations of mineralelements and storage protein composition in the matured seeds were analyzed. Non-nodulating (cv T201) and nodulating (cv Enrei) soybean plant were cultivated with nitrate (NaNO3) and fix-N as solo nitrogen source, respectively. Application of phosphate increased P and reduced N concentration in seeds both non-nodulating and nodulating soybean plants. This result indicated that seed mineral composition could be changed by phosphate application irrespective of nitrogen source. Low level phosphate (5μM-P) application to the non-nodulating soybean plants increased β-conglycinin, a major storage protein in soybean seed, concentration in seed compared with normal level of P (25μM-P) irrespective of nitrate application level. In the nodulating soybean plants, the concentration of β-subunit of β-conglycinin was decreased by high level of P (50μM-P) application both all through plants growth and increasing P level after flowering. In conclusion, P application on soybean plants changed mature seed mineral composition and protein composition irrespective of N nutrition.ダイズをさまざまなリン濃度で水耕栽培し、種子の各種元素濃度および貯蔵タンパク質組成を比較した。根粒非着生系統ダイズ(品種T201)と栽培ダイズ(品種エンレイ)をそれぞれ、硝酸あるいは窒素固定のみを唯一の窒素限として栽培した。両植物とも、植物体に対するリン供給量の増加は、種子におけるリン濃度を増加させ、窒素濃度を減少させた。この結果は、種子に貯蔵される元素濃度は、窒素の供給形態に関わらず、リン供給により変化することを示している。根粒非着生系統ダイズに対する低濃度のリンの供給(5μM-P)は、対照区(25μM-P)と比較して、窒素の供給量によらず、種子貯蔵タンパク質のβ- コングリシニン含量を増加させた。栽培系統ダイズのエンレイでは、β- コングリシニンのβサブユニット濃度は、生育全期間あるいは開花期から高濃度のリンの供給(50μM-P)により減少した。 これらの結果から、ダイズにおける種子の元素集積および貯蔵タンパク組成は窒素の供給形態に関わらず、リンの供給量により変えることができることが示された。

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    その他リンク: http://hdl.handle.net/10191/47174

  • 39 水耕ダイズに対する無機態窒素供給が根粒肥大と培地pHに与える影響(関東支部講演会,2014年度各支部会)

    小野 雄基, 後藤 大輝, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 61 )   266 - 266   2015年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • P4-1-9 オオムギ高親和性硝酸輸送系の制御機構(ポスター,4-1 植物の多量栄養素,2015年度京都大会)

    篠原 雅人, 大竹 憲邦, 大山 卓爾, 末吉 邦

    日本土壌肥料学会講演要旨集   ( 61 )   61 - 61   2015年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • P4-1-6 チューリップの導管中の窒素の移動形態について(ポスター,4-1 植物の多量栄養素,2015年度京都大会)

    堤 洸太郎, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 61 )   60 - 60   2015年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • P4-4-5 短時間の無機態窒素供給によるダイズ体内成分の変動(ポスター,4-4 植物の代謝成分と農作物の品質,2015年度京都大会)

    小野 雄基, 後藤 大輝, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 61 )   87 - 87   2015年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • ダイズの窒素固定依存率迅速測定のための微量相対ウレイド分析法

    大山 卓爾, 坂詰 大志, 石川 伸二, 末吉 邦, 大竹 憲邦, 高橋 能彦, 南雲 芳文

    農業および園芸   89 ( 10 )   977 - 987   2014年10月

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    記述言語:日本語   出版者・発行元:養賢堂  

    ダイズは,豆腐,味噌,醤油,納豆など我が国の伝統食品の原料作物であるとともに,世界的にも油料および飼料用穀物として生産が増加している。我が国ではダイズは年間約400万トンが消費されているが,国内生産高は約20万トンと低く,平均収量も10aあたり170kg(農林水産省大豆関連データ)と世界平均248kg(FAOSTAT 2013)と比べても低迷している。ダイズは種子にタンパク質を約35-40%と高濃度に含み,開花期以降も多量の窒素を必要とする(大山 2001)。ダイズは,根に土壌微生物である根粒菌を共生し,空中窒素を固定利用できる。また,ダイズは他の作物と同様に肥料窒素や土壌窒素も利用できるが,速効性窒素肥料を多量に施用すると,根粒の着生および窒素固定活性を抑制することが知られている。したがって,ダイズの施肥や栽培技術の改善のためには,根粒による窒素固定と根からの化合態窒素の吸収割合を調べることにより有用な情報が得られる。

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    その他リンク: http://agriknowledge.affrc.go.jp/RN/2010873737

  • 導管液と茎アポプラスト液の微量相対ウレイド分析によるダイズの窒素固定依存率の推定

    坂詰 大志, 田中 一也, 會田 浩基, 石川 伸二, 南雲 芳文, 高橋 能彦, 大竹 憲邦, 末吉 邦, 大山 卓爾, 坂詰 大志, 田中 一也, 會田 浩基, 石川 伸二, 南雲 芳文, 高橋 能彦, 大竹 憲邦, 末吉 邦, 大山 卓爾

    新潟大学農学部研究報告   67 ( 1 )   27 - 41   2014年9月

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    記述言語:英語   出版者・発行元:新潟大学農学部  

    相対ウレイド法は、圃場栽培ダイズの窒素固定依存率を推定するための簡便で信頼のできる方法である。一般に主茎株切断面から溢泌する導管液中の硝酸、アミド、ウレイド濃度を分光光度計を用いて比色定量する。我々は、通常の分析で用いる導管液量50μLの代わりに、その1/20の2.5μLを用いる微量分析法を開発した。呈色反応は、1.5mLエッペンドルフチューブ内で行ない、呈色液200μLを96穴マイクロプレートのウェルに入れ、マイクロプレートリーダーで吸光度を測定した。標準液を2連で測定することにより、精度の高い検量線の作成が可能であった。本分析法を用いることにより、1日に200試料を測定できた。また、生育後期(黄葉期R7)や、土壌が乾燥すると、導管液を溢泌しないことがあるため、確実に試料を採取する方法として、携帯型遠心器(2,200 x G,10秒)を用いて茎のアポプラスト液を採取した。茎は、主茎下部から2,3cmの長さで切り出し、1.5mLのエッペンドルフチューブに入れて回収した。茎のアポプラスト液は、時期や気象条件によらず株あたり30-50mg程度回収できた。各窒素成分濃度および相対ウレイドパーセントを比較したところ、R7では、導管液とほぼ同様の値が得られ、導管液の代わりに利用できる可能性が示された。しかしながら、R1,R5では、アポプラスト液のアミド,硝酸濃度が導管液よりも高く、相対ウレイドパーセントは、50%程度と導管液80-90%と異なる値となった。

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    その他リンク: http://hdl.handle.net/10191/30053

  • ダイズ根粒の生長におよぼす温度と明暗の影響

    小野雄基, 石川慶祐, 石川伸二, 大竹憲邦, 末吉邦, 七夕小百合, 七夕高也, 大山卓爾

    植物微生物研究会研究交流会講演要旨集   24th   2014年

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  • ダイズ根粒成長解析のためのコンピュータ画像計測方法

    七夕小百合, 七夕高也, 齋藤明徳, 田嶋誠也, 渡辺詩織, 石川慶祐, 大竹憲邦, 末吉邦, 大山卓爾

    日本土壌肥料学雑誌   85 ( 1 )   2014年

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  • 施肥窒素供給期間によるサトウキビ(Saccharum officinarum L.)の生育に対する窒素固定寄与の15N希釈法による推定

    飛山 隆洋, 百瀬 篤志, 西村 圭子, 石崎 記子, 石川 伸二, 山本 美祥, Hung N.V.P., Chamaiporn A., Ruamrungsri S., 大竹 憲邦, 末吉 邦, 大山 卓爾

    新潟大学農学部研究報告 = Bulletin of the Faculty of Agriculture, Niigata University   66 ( 1 )   11 - 19   2013年10月

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    記述言語:英語   出版者・発行元:新潟大学農学部  

    施肥窒素供給期間の違いによるサトウキビ(Saccharum officinarum L., 品種農林8号)の窒素固定寄与率を15N希釈法で調べる目的で新潟大学の温室内でポット試験を実施した。サトウキビの種茎を水中で発芽させたのち,バーミキュライトを充填した1/5000aのワグネルポットで栽培した。窒素施肥処理は,3区設けた。1)N0区:無窒素培養液のみ供与,2)N100:15N標識の硫酸アンモニウム水溶液(100mgN)を週1回ポットに添加,3)N100N0:移植から移植後6週目まで,15N標識の硫酸アンモニウム水溶液(100mgN)を週1回ポットに添加し,その後無窒素栽培した。移植後植物の地上部の長さと葉数を毎週測定した。また,移植12週間後と20週間後に植物を採取し,各部位の窒素含有量,15N濃度とアセチレン還元活性を測定した。N0区の植物はほぼ生長が停止し,窒素固定だけでは生育を支えるには不十分であることが確認された。N100区とN100N0区の植物は,地上部の長さや葉の数は,大きな差はなかったが,N100N0区の葉は葉色が薄く,窒素含有量もN100区の半分程度であった。移植12週間後には,N100区とN100N0区の株あたり全窒素含有率は,それぞれ408mgNと286mgNであった。15N希釈法で推定した窒素固定量は,N100区とN100N0区でそれぞれ株あたり87mgN(窒素固定由来窒素割合%Ndfa,21%),48mgN(17%Ndfa)であった。移植20週間後では,N100区とN100N0区の株あたり全窒素含有率は,それぞれ569mgNと292mgNであった。窒素固定推定量は,N100区とN100N0区でそれぞれ株あたり87mgN(%Ndfa,21%),57mgN(33%Ndfa)であった。器官別窒素固定割合では,枯葉,種茎が高く,緑葉,茎が低い傾向がみられた。

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  • ミヤコグサ根粒超着生変異株har1-4の葉の生育

    七夕 小百合, 大竹 憲邦, 末吉 邦, 川口 正代司, 大山 卓爾, 七夕 小百合, 大竹 憲邦, 末吉 邦, 川口 正代司, 大山 卓爾

    新潟大学農学部研究報告   66 ( 1 )   21 - 24   2013年10月

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    記述言語:英語   出版者・発行元:新潟大学農学部  

    ダイズの根粒超着生変異株は親株と比較し,葉面積が小さく,葉の細胞数が少ない。そのため,ダイズの根粒形成自己制御機構は葉の細胞増殖と関連することが示唆されている。本報告では,マメ科モデル植物であるミヤコグサの根粒超着生変異株har1-4の葉の生育を親株と比較した。har1-4の葉面積は,親株と同程度であった。har1-4の葉の細胞数は親株より少ない傾向にあり,har1-4の細胞面積は,親株よりも大きい傾向にあった。以上の結果から,ミヤコグサにおいても根粒形成自己制御機構は葉の細胞増殖と関連する可能性が示唆された。マメ科植物の根粒形成自己制御機構は,根粒形成に特異的な機構というよりは,植物の一般的なメカニズムの一部を利用した機構であるのかもしれない。

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    その他リンク: http://hdl.handle.net/10191/23967

  • Estimation of contribution of nitrogen fixation in sugarcane (Saccharum officinarum L.) plants with ¹⁵N dilution method in relation to nitrogen supply period

    飛山 隆洋, 百瀬 篤志, 西村 圭子, Ishizaki Noriko, Ishikawa Shinji, Yamamoto Misaki, Hung Ngyuen Van Phi, Chamaiporn Anuwong, Ruamrungsri Soraya, Ohtake Norikuni, Sueyoshi Kuni, Ohyama Takuji, 飛山 隆洋, 百瀬 篤志, 西村 圭子, 石崎 記子, 石川 伸二, 山本 美祥, 大竹 憲邦, 末吉 邦, 大山 卓爾

    新潟大学農学部研究報告   66 ( 1 )   11 - 19   2013年10月

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    記述言語:英語   出版者・発行元:新潟大学農学部  

    Pot experiment was conducted in warm greenhouse in Niigata University to estimate the contribution of nitrogen fixation in sugarcane (Saccharum officinarum L. var. NiF8) plants using 15N dilution method in relation to N supply period. Sugarcane plants were grown from a cut stalk in water for 20 days and the young shoots were transplanted to the 1/5000 a Wagner pot filled with vermiculite. Three fertilizer treatments were applied, 1) N0: Nitrogen free culture solution was supplied, 2) N100: 15N labeled ammonium sulfate was supplied at the rate of 100 mgN per pot a week, 3) N100N0: 15N labeled ammonium sulfate was supplied at the rate of 100 mgN per por a week until 6 weeks after transplanting (WAT), then the plants were cultivated without N supply. The growth of the plants was measured every week, and plants were harvested at 12 WAT and 20 WAT. Total nitrogen content and 15N abundance in each part were determined. Acetylene reduction activity was measured for each part of harvested plants. N0 plants grew very poor, and sole nitrogen fixation is not enough to support vigorous growth of sugarcane plants. N100 plants and N100N0 plants showed relatively similar shoot length and leaf number, but the N100N0 plants leaves were pale and the N concentration was almost a half of N100 plants. At 12 WAT, total N content was 408 mgN and 286 mgN per plant in N100 treatment and N100N0 treatment, respectively. The amount of nitrogen derived from nitrogen fixation (Ndfa) in N100 and N100N0 was 87 mgN (21%Ndfa) and 48 mgN (17%Ndfa) per plant respectively. At 20 WAT, total N content was 569 mgN (100N) and 292mgN (100N0N), and the amount of Ndfa was 87 mg (15%Ndfa) in 100N and 57 mgN (20%Ndfa) in 100N0N treatment. Among organs, the estimated %Ndfa tended to be higher in old leaves and stalk, and lower in green leaves and stems. From this experiment, the continuous supply of N fertilizer did not inhibit nitrogen fixation in sugarcane compared with N deficient plants.施肥窒素供給期間の違いによるサトウキビ(Saccharum officinarum L., 品種農林8号) の窒素固定寄与率を15N希釈法で調べる目的で新潟大学の温室内でポット試験を実施した。サトウキビの種茎を水中で発芽させたのち、バーミキュライトを充填した1/5000 a のワグネルポットで栽培した。窒素施肥処理は、3区設けた。1) N0区: 無窒素培養液のみ供与、 2) N100: 15N標識の硫酸アンモニウム水溶液(100mgN)を週1回ポットに添加、3) N100N0: 移植から移植後6週目まで、15N標識の硫酸アンモニウム水溶液(100mgN)を週1回ポットに添加し、その後無窒素栽培した。移植後植物の地上部の長さと葉数を毎週測定した。また、移植12週間後と20週間後に植物を採取し、各部位の窒素含有量、15N濃度とアセチレン還元活性を測定した。N0区の植物はほぼ生長が停止し、窒素固定だけでは生育を支えるには不十分であることが確認された。N100区とN100N0 区の植物は、地上部の長さや葉の数は、大きな差はなかったが、N100N0区の葉は葉色が薄く、窒素含有量もN100区の半分程度であった。移植12週間後には, 100 区とN100N0 区の株あたり全窒素含有率は、それぞれ408 mgN と286 mgN であった。15N 希釈法で推定した窒素固定量は、N100 区とN100N0 区でそれぞれ株あたり87mgN(窒素固定由来窒素割合%Ndfa, 21%)、48mgN(17%Ndfa)であった。移植20週間後では、N100区とN100N0区の株あたり全窒素含有率は、それぞれ569mgNと292mgNであった。窒素固定推定量は、N100区とN100N0区でそれぞれ株あたり87mgN(%Ndfa, 21%)、57mgN(33% Ndfa)であった。器官別窒素固定割合では、枯葉、種茎が高く、緑葉、茎が低い傾向がみられた。

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    その他リンク: http://hdl.handle.net/10191/23964

  • サトウキビ(Saccharum officinarum L.)から分離した窒素固定内生菌の窒素固定と窒素放出機構

    百瀬 篤志, 飛山 隆洋, 西村 圭子, 石崎 記子, 石川 伸二, 山本 美祥, Hung N.V.P., 大竹 憲邦, 末吉 邦, 大山 卓爾

    新潟大学農学部研究報告 = Bulletin of the Faculty of Agriculture, Niigata University   66 ( 1 )   1 - 9   2013年10月

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    記述言語:英語   出版者・発行元:新潟大学農学部  

    サトウキビ(Saccharum officinarum L., 品種農林8号)の茎から分離された内生窒素固定菌(JA1株,JA2株,菌の性質からGluconacetobacter diazotrophicusと予測された)の15N2固定活性とアセチレン還元活性(ARA) を調べた。試験管内で固形LGIP培地または液体LGIP培地で培養した菌を,酸素濃度を0から20%まで変化させて,24時間15N2固定活性を測定し,その後同じ試験管で1時間ARAを測定した。固形培地では,O2 0.4%で最大活性を示したが,液体培地では,O2 0%で最大活性を示した。どちらも,O2 20%まで,酸素濃度が高まるにつれて活性が低下した。JA1株の生育密度(吸光度)とARAをLGIP液体培地に植継いでから10日後まで毎日測定した。菌密度は,植継1日後に急速に高まり6日後までは増加したが,それ以降10日目までは増加が抑制された。JA1株の増殖抑制はクオラムセンシング機構によると予想される。ARAも植継5日目までは上昇したが,6日目以降急激に低下した。7日目から10日目までは,ほとんど活性を示さなかった。この結果は,本菌の窒素固定活性は,菌の増殖中にのみ発現し,増殖が止まると窒素固定活性も停止することが示唆された。このことが,サトウキビ植物体内でも同様であるとすると,内生菌が増殖を続けることが,窒素固定の維持に必要であると推定される。内生菌から,植物への窒素の放出は,サトウキビの生育促進に重要である。液体培養したJA1株は,24時間の15N2固定直後では,大部分の固定窒素が菌体内に残存しており,培地中には,固定窒素総量の4%程度のみ放出していた。一方,24時間15N2を固定させた後,酸素濃度0-20%で10日間培養を続けたところ,酸素濃度が高い程,放出15N量の割合が増加し,O2 20%では,固定窒素総量の40%が培地に放出されていた。これらの結果は,JA1株は,窒素固定停止後,増殖を停止し,菌の死に伴う分解により,窒素を放出するのではと予想された。

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  • P7-1-3 窒素深層施肥による水田フル活用 : 新潟県畑土壌におけるオオムギへの窒素深層施肥効果(ポスター,シンポジオン,7-1 肥料および施肥法,2013年度名古屋大会)

    末吉 邦, 石川 伸二, 服部 誠, 藤田 与一, 南雲 芳文, 浦野 保徳, 大竹 憲邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 59 )   137 - 137   2013年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • 7-1-36 石灰窒素の土壌中での動態解析(7-1 肥料および施肥法,2013年度名古屋大会)

    大竹 憲邦, 石黒 竜成, 山本 美祥, 石川 伸二, 南雲 芳文, 服部 誠, 高橋 能彦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 59 )   135 - 135   2013年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • P4-1-9 オオムギにおける二成分高親和性硝酸輸送系の解析(4-1 植物の多量栄養素)

    高田 雅大, 石川 伸二, 大竹 憲邦, 大山 卓爾, 末吉 邦

    日本土壌肥料学会講演要旨集   ( 59 )   64 - 64   2013年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • ダイズ根粒の生長におよぼす温度と日周期の影響

    石川慶祐, 渡辺詩織, 七夕高也, 七夕小百合, 石川伸二, 大竹憲邦, 大竹憲邦, 末吉邦, 末吉邦, 大山卓爾, 大山卓爾

    植物微生物研究会研究交流会講演要旨集   22nd   2013年

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  • P12-1 チューリップ根のグルタミン合成酵素の解析(12.植物の代謝成分・農作物の品質,2012年度鳥取大会)

    山本 美祥, 堤洸 太郎, 会田 知夏, 種村 竜太, 石川 伸二, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 58 )   90 - 90   2012年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • ダイズ根粒の生長におよぼす硝酸,温度,明暗条件の影響について

    渡辺詩織, 七夕小百合, 七夕高也, 田嶋誠也, 大竹憲邦, 末吉邦, 大山卓爾

    植物微生物研究会研究交流会講演要旨集   21st   2012年

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  • 12-4 ダイズ栽培における窒素及びリン酸質肥料の施用効果(12.植物の代謝成分と農作物の品質)

    大竹 憲邦, 七夕 小百合, 山本 美祥, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 57 )   93 - 93   2011年8月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • P9-8 シロイヌナズナ硝酸低蓄積突然変異体の解析(ポスター紹介,9.植物の多量栄養素,2010年度北海道大会)

    太田 弘毅, 星野 はるな, 石川 伸二, 川地 太平, 原田 久富美, 大竹 憲邦, 大山 卓爾, 末吉 邦

    日本土壌肥料学会講演要旨集   ( 56 )   65 - 65   2010年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • 明暗期におけるダイズ根粒と根の生長に及ぼす硝酸の効果

    齋藤明徳, 伊藤小百合, 七夕高也, 田嶋誠也, 大竹憲邦, 末吉邦, 大山卓爾

    植物微生物研究会研究交流会講演要旨集   19th   2010年

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  • P8-15 導管液を用いたダイズの根粒窒素固定活性測定と無機栄養診断技術の開発(ポスター紹介,8.共生,2008年度愛知大会)

    坂詰 大志, 高橋 能彦, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 54 )   69 - 69   2008年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • P19-10 緩効性窒素肥料の深層施肥がダイズ子実の収量および品質に及ぼす影響(ポスター紹介,19.肥料および施肥法,2008年度愛知大会)

    田中 一也, Tewari Kaushal, 大竹 憲邦, 末吉 邦, 高橋 能彦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 54 )   157 - 157   2008年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • P10-4 シロイヌナズナ硝酸低蓄積突然変異体の解析(ポスター紹介,10.植物の代謝,2008年度愛知大会)

    星野 はるな, 石橋 弘規, 川地 太兵, 原田 久富美, 大竹 憲邦, 大山 卓爾, 末吉 邦

    日本土壌肥料学会講演要旨集   ( 54 )   102 - 102   2008年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • P8-14 硝酸によるダイズ根粒の肥大成長阻害について : 光および根の伸長との関連(ポスター紹介,8.共生,2008年度愛知大会)

    齋藤 明徳, 木村 拓也, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 54 )   69 - 69   2008年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • 8-9 ダイズ葉柄から供与した植物ホルモンの移動と根粒形成に及ぼす影響(8.共生,2007年度東京大会)

    種田 祐士, 伊藤 小百合, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 53 )   54 - 54   2007年8月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • P10-11 シロイヌナズナ硝酸蓄積突然変異体の解析(10.植物の代謝,2007年度東京大会)

    石橋 弘規, 川地 太兵, 原田 久富美, 大竹 憲邦, 大山 卓爾, 末吉 邦

    日本土壌肥料学会講演要旨集   ( 53 )   98 - 98   2007年8月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • P10-10 オオムギ低親和性硝酸トランスポーターの機能解析(10.植物の代謝,2007年度東京大会)

    小野 大輔, 石川 伸二, 大竹 憲邦, 大山 卓爾, 末吉 邦

    日本土壌肥料学会講演要旨集   ( 53 )   98 - 98   2007年8月

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  • P8-1 大豆に共生する根粒菌及びAzospirillum菌の動態及び大豆の成長と窒素固定に与える影響について(8.共生,2007年度東京大会)

    本間 史生, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 53 )   60 - 60   2007年8月

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  • P11-8 ^<15>N analysis of deep placement of lime nitrogen at various growth stages of soybean (Glycine max (L) merr.) plants

    Tewari Kaushal, 佐藤 隆紀, 佐藤 直美, 高橋 能彦, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 52 )   99 - 99   2006年9月

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    記述言語:英語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • 8-13 ダイズ根粒超着生変異株NOD系統の表現型の比較(8. 共生, 2006年度秋田大会講演要旨)

    伊藤 小百合, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 52 )   45 - 45   2006年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • 9-58 シロイヌナズナにおける硝酸低蓄積型突然変異体の解析(9. 植物の無機栄養, 2006年度秋田大会講演要旨)

    石橋 弘規, 石川 伸二, 今野 公博, 星野 はるな, 川地 太兵, 原田 久富美, 大竹 憲邦, 大山 卓爾, 末吉 邦

    日本土壌肥料学会講演要旨集   ( 52 )   70 - 70   2006年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • 9-61 ダイズ種子貯蔵タンパク質転写調節に関与するSWI/SNF様タンパク質の解析(9. 植物の無機栄養, 2006年度秋田大会講演要旨)

    大竹 憲邦, 伊藤 小百合, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 52 )   71 - 71   2006年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • P8-10 ダイズ葉柄から供与したオーキシンの根粒形成に及ぼす影響(8. 共生, 2006年度秋田大会講演要旨)

    種田 祐士, 伊藤 小百合, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 52 )   49 - 49   2006年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • P8-11 ダイズ品種 Williams と根粒超着生変異株NOD 1-3に下部根から与えた硝酸が生育と窒素固定に与える影響(8. 共生, 2006年度秋田大会講演要旨)

    木村 拓也, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 52 )   49 - 49   2006年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • ダイズ根粒超着生変異株NOD1-3の根粒形成初期過程における光合成産物の分配

    Ito Sayuri, Ohtake Norikuni, Sueyoshi Kuni, Ohyama Takuji, 伊藤 小百合, 大竹 憲邦, 末吉 邦, 大山 卓爾

    新潟大学農学部研究報告   59 ( 1 )   33 - 38   2006年8月

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    記述言語:英語   出版者・発行元:新潟大学農学部  

    We reported previously that photosynthate allocation for nodule formation did not increase during early stage of nodule formation in soybean (Glycine max [L.] Merr.) cv. Williams (Ito et al., 2006). In this study, current photosynthate allocation of hypernodulation mutant of soybean NOD1-3 isolated from Williams was examined in early stage of nodule formation. Whole shoots were exposed to 14CO2 for 120 min and the distribution of radioactivity in each organ was determined. The 14C distribution in the roots of 8 days after inoculation did not increase when compared with uninoculated control plants. In visualized images of radioactivity by imaging plate, nodules were observed as strong signal spots in the underground organ. These results indicate that current photosynthate allocation to the inoculated root did not increase markedly during the early stages of nodule formation, while small nodules had already strong sink activity than root in 8 days after inoculation different from Williams. To investigate the carbon status of plants at 8 days after inoculation, the starch and sugar concentration in the plants were analyzed. It was shown that the accumulation of starch and sugar was similar in both inoculated and uninoculated plants. We concluded that appreciable amount of photoassimilate is not required for nodule initiation in NOD1-3 in 8 days after inoculation.これまでに、ダイズ(Glycine max [L] Merr.)栽培品種Williamsを用い、根粒菌接種8日後までの根粒形成初期過程では、根粒着生による光合成産物の分配増加が認められないことを示した(Ito et al. 2006)。今回は、Williamsより得られた根粒超着生変異株NOD1-3を用い、根粒形成初期過程における光合成産物の要求性に違いがあるか否かを調査した。播種18日後のダイズ植物の地上部に14CO2を120分間供給する実験を行った。根粒菌接種8日後の地下部への14Cの分配を、根粒菌非接種の地下部への分配と比較した。接種8日後では根粒超着生変異株においても、接種した根への光合成産物の分配の増加は認められなかった。しかしイメージングプレートを用いた14C分配のイメージング画像では、Williamsとは異なり接種した地下部において根粒の部分が強いシグナルとして検出された。接種8日後では地下部に送られる光合成産物量全体は非接種と変わらないが、局所的には根粒が根よりもすでに強いシンク活性を持つことを示した。また、より長期的な光合成産物の動態を調査するために、植物体内のデンプンおよび糖含量を分析した。しかし、デンプン・糖含量ともに根粒菌接種・非接種処理間に有意な差は無かった。これらの結果から、NOD1-3においても根粒形成初期過程における光合成産物の地下部への分配は顕著に増加することはないと結論づけた。

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  • P11-6 Comparison of deep placement of slow release fertilizer (lime nitrogen) on yield and quality of soybean (Glycine max (L.) Merr.) seeds at various depth and with different amounts

    Tewari Kaushal, 遠田 優, 佐藤 直美, 伊藤 小百合, 藤掛 浩行, 大竹 憲邦, 末吉 邦, 高橋 能彦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 51 )   103 - 103   2005年9月

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    記述言語:英語   出版者・発行元:一般社団法人日本土壌肥料学会  

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  • P9-1 シロイヌナズナ硝酸蓄積突然変異体の解析(ポスター紹介,9.植物の無機栄養,日本土壌肥料学会 2005年度大会講演要旨集)

    石橋 弘規, 川地 太兵, 原田 久富美, 大竹 憲邦, 大山 卓爾, 末吉 邦

    日本土壌肥料学会講演要旨集   ( 51 )   78 - 78   2005年9月

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  • 10-8 ダイズ種子貯蔵タンパク質の窒素応答性機構の解析(10.植物の代謝,日本土壌肥料学会 2005年度大会講演要旨集)

    大竹 憲邦, 伊藤 小百合, 大友 渉, 楠 佳輝, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 51 )   88 - 88   2005年9月

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  • ダイズ根粒乾燥誘導遺伝子 (ERD15) の硝酸応答性(8. 共生, 2004年度大会講演要旨集)

    山崎 明彦, 藤掛 浩行, 伊藤 小百合, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 50 )   62 - 62   2004年9月

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  • ダイズ根粒形成初期過程における光合成産物の分配(8. 共生, 2004年度大会講演要旨集)

    伊藤 小百合, 大竹 憲邦, 山崎 明彦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 50 )   62 - 62   2004年9月

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  • ダイズ未熟子葉の試験管培養における培地成分が種子貯蔵タンパク質βコングリシニンのβサブユニットmRNA集積に及ぼす影響

    大竹 憲邦, 岡野 小百合, 末吉 邦, 大山 卓爾

    新潟大学農学部研究報告   57 ( 1 )   41 - 45   2004年8月

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    記述言語:日本語   出版者・発行元:新潟大学  

    ダイズ種子貯蔵タンパク質の一つであるβコングリシニンのβサブユニットの集積調節物質を検索するために、未熟子葉の試験管培養系を用いていくつかのアミノ酸、植物ホルモン及び有機酸添加に対する応答性を調査した。グルタミン酸、アラニンを添加すると、未熟子葉内の遊離アミノ酸が上昇し、いくつかのアミノ酸が増加したことから、これらのアミノ酸は種子に取り込まれ、代謝されたことが示唆された。しかし、βサブユニットmRNAはグルタミン添加培地以外では検出できなかった。アスパラギン酸添加では未熟子葉内の遊離アミノ酸濃度の上昇及びβサブユニットmRNAの集積は認められなかった。植物ホルモンの一つサイトカイニン(t-zeatin)の添加によってもβサブユニットの集積は促進されなかった。TCA回路のα-ケトグルタル酸、オキサロ酢酸、クエン酸の添加によってもβサブユニットmRNAの集積は促進されなかった。また培養後の未熟子葉内におけるこれらの有機酸濃度にはグルタミンのみ添加の対照区と比較して有意な上昇は認められなかった。以上の実験結果から、ダイズ未熟子葉において少なくとも今回試験に用いた物質の子葉外濃度の上昇によりβサブユニットmRNAの集積は促進されないことが確認された。It has been reported that the accumulation of soybean (Glycine max L.) storage protein, β subunit of β conglycinin, was suppressed by nitrogen (N)-deficiency. In this report we tried to evaluate what compound(s) may have a key role in regulating the accumulation of β subunit mRNA in vitro cotyledon culture system. The cotyledons from non-nodulating soybean plants cultivated under N-deficient condition (0.5 mM NO 3 -) were transferred to in vitro culture system. The application of glutamine (Gln) enhanced β subunit mRNA accumulation within twelve hours. The application of glutamic acid, alanine, or aspertic acid could not induced β subunit mRNA accumulation. Application of cytokinin (t-zeatin) oxaloasetate, 2 -oxoglutarate and citrate could not enhance the accumulation of β subunit mRNA under N free culture medium. These results suggested that at the least application to theses amino acids, orgamic acids and cytokinin could not be a direct signal to enhance β subunit mRNA accumulation in invitro cotyledon culture system.

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    その他リンク: http://agriknowledge.affrc.go.jp/RN/2010700658

  • 9-44 オオムギにおけるアニオンチャネルホモログの発現解析(9.植物の無機栄養)

    北林 雅士, 川地 太兵, 大竹 憲邦, 大山 卓爾, 末吉 邦

    日本土壌肥料学会講演要旨集   ( 49 )   79 - 79   2003年8月

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  • 10-15 ダイズ植物体内における窒素化合物の移動と分配(10.植物の代謝)

    大竹 憲邦, 高野 史, 藤掛 浩行, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 49 )   94 - 94   2003年8月

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  • 窒素深層施肥と根粒菌接種が客土造成転換畑初作ダイズに及ぼす影響

    Kaushal Tewari, 皆川 律子, 菅沼 丈人, 藤掛 浩行, 大竹 憲邦, 末吉 邦, 高橋 能彦, 土田 徹, 大山 卓爾

    日本土壌肥料學雜誌 = Journal of the science of soil and manure, Japan   74 ( 2 )   183 - 189   2003年4月

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    記述言語:日本語   出版者・発行元:日本土壌肥料学会  

    GUS標識根粒菌を用い,バーミキュライトを含む5種類の土壌に接種した根粒菌の増殖を調べたところ,どの土壌でも接種1週間後に根粒菌密度は10倍以上に増加した。そこで,尿素,被覆尿素,石灰窒素の深層施肥区にバーミキュライトを充填したペーパーポットに播種と同時に根粒菌を接種し初生葉展開時に移植する方法,種子接種苗移植またはペーパーポット無接種苗を移植する方法で栽培を行った。土着根粒菌が定着していない建設残土を上乗せ客土した新潟県内の大規模水田転換畑初作圃場内で窒素肥料の深層施肥試験を行った。施肥は,標準施肥(1.6g N m^<-2>)に加え,尿素,100日タイプの被覆尿素,または石灰窒素(10g N m^<-2>)を移植直下深さ20cmの位置に深層施肥し,ダイズの子実収量と窒素含有量に対する施肥と根粒菌接種の効果を調べた。根粒菌無接種ペーパーポット苗移植では根粒の着生が認められず,窒素深層施肥無施用区の子実収量は約9g株^<-1>ともっとも低かった。無施用区のペーパーポット接種苗移植,種子接種苗移植ともに子実収量は約22g株^<-1>となり,根粒菌接種で有意に高まった。葉の寿命は石灰窒素区でもっとも長かった。また,株あたり子実収量は,尿素区32〜37g,被覆尿素区40〜44g,石灰窒素区38〜48gと深層施肥により増加した。以上の結果から,根粒菌を接種した苗を移植栽培することにより,根粒菌のいない転換畑の初年作でも根粒形成と窒素固定の寄与が期待できること,および,石灰窒素の深層施肥はダイズの生育および収量の増加に被覆尿素と同等の効果があることが示唆された。Population of GUS marked bradyrhizobia inoculated in five different soils including calcined vermiculite increased over 10 times in a week. Each soybean seed was planted in a paper pot filled with vermiculite inoculated with or without bradyrhizobia, and the 10-d-old seedlings were transplanted with a paper pot to the first cropping reclaimed field in Niigata. In addition to the conventional fertilization (1.6 g N m^<-2>), deep placement of urea, 100 d type coated urea, or calcium cyanamide (10 g N m^<-2>) was applied. In conventional fertilization the soybean plants transplanted with non-inoculated paper pot did not nodulate due to the absence of indigenous bradyrhizobia, and the seed dry weight was lowest, about 9 g plant^<-1>. Seed dry weight of the plants either with paper pot inoculation or seed inoculation was higher, about 22 g plant^<-1>. Soybean plants with deep placement of calcium cyanamide exhibited prolonged leaf activity. With deep placement of urea, coated urea and calcium cyanamide, seed dry weight increased to 32-37, 40-44 and 38-47g plant^<-1> respectively. The highest average seed yield was obtained in the inoculated paper pot transplantation with calcium cyanamide.

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  • 6 Comparative studies on contribution of roots and shoots to in vivo nitrate reduction in wild type and nitrate reductase-deficient mutant of barely

    Latif Salwa, Kawachi Tahei, Fujikake Hiroyuki, Ohtake Norikuni, Sueyoshi Kuni, Ohyama Takuji

    日本土壌肥料学会講演要旨集   ( 48 )   212 - 212   2002年3月

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  • 9-51 オオムギおよびレタスにおけるアニオンチャネルの発現解析(9.植物の無機栄養)

    末吉 邦, 川地 太兵, 藤掛 浩行, 大竹 憲邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 48 )   62 - 62   2002年3月

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  • 10-5 ダイズ種子貯蔵タンパク質集積に及ぼすアミノ酸の影響(10.植物の代謝)

    大竹 憲邦, 末吉 邦, 川地 太兵, 藤掛 浩行, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 48 )   72 - 72   2002年3月

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  • 9-52 オオムギ根における硝酸トランスポーターの発現解析(9.植物の無機栄養)

    石川 伸二, 川地 太兵, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 48 )   63 - 63   2002年3月

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  • 8-18 ダイズ根粒の肥大生長における硝酸の影響とその阻害機構(8.共生)

    藤掛 浩行, 菅沼 丈人, 田村 洋平, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 48 )   42 - 42   2002年3月

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  • 現代土壌肥料学の断面(4)植物硝酸還元酵素の機能発現制御のしくみ

    末吉 邦, 川地 太兵

    農業および園芸   76 ( 4 )   509 - 515   2001年4月

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    記述言語:日本語   出版者・発行元:養賢堂  

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  • 9-46 オオムギにおける硝酸イオンの輸送に対するアニオンチャネル阻害剤の影響(9.植物の無機栄養)

    末吉 邦, 川地 太岳, 西條 千鶴, 大竹 憲邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 47 )   83 - 83   2001年3月

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  • 21-13 砂質土壌ポット栽培レタスに対する醤油粕の施肥効果(21.土壌改良資材)

    藤井 邦彦, 藤原 和成, 大竹 憲邦, 末吉 邦, 大山 卓爾, 大辻 英郎

    日本土壌肥料学会講演要旨集   ( 47 )   190 - 190   2001年3月

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  • 9-45 オオムギアニオンチャネル遺伝子のクローニングと機能解析(9.植物の無機栄養)

    川地 太岳, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 47 )   83 - 83   2001年3月

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  • 土壌中の可給態窒素に応答する植物機能

    末吉 邦, 川地 太兵, 大竹 憲邦, 大山 卓爾

    日本農芸化学会誌   75   463 - 463   2001年3月

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  • 8-16 ダイズ根粒の硝酸阻害と光合成産物の関係について(8.共生)

    藤掛 浩行, 菅沼 丈人, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 46 )   60 - 60   2000年3月

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  • 9-19 オオムギ高親和性硝酸トランスポーターの発現制御機構に関する研究(9.植物の無機栄養)

    菊地 章子, 川地 太兵, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 46 )   75 - 75   2000年3月

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  • 10-5 オオムギ硝酸還元酵素の誘導に及ぼすMSXの影響(10.植物の代謝)

    高橋 泉, 川地 太兵, 大竹 憲邦, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 46 )   96 - 96   2000年3月

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  • 10-13 窒素によるダイズ種子貯蔵タンパク質の調節機構の解析(10.植物の代謝)

    大竹 憲邦, 佐藤 明世, 奥山 郁子, 末吉 邦, 大山 卓爾

    日本土壌肥料学会講演要旨集   ( 46 )   100 - 100   2000年3月

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  • INACTIVATION OF BARLEY NITRATE REDUCTASE BY 14-3-3 PROTEIN

    SUEYOSHI Kuni, HAYASHI Yuko, OHTAKE Norikuni, SUGIMOTO Toshio, OJI Yoshikiyo, OHYAMA Takuji

    Plant and cell physiology   40   s139 - s139   1999年3月

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  • Measurement Method of Gytokinis in Soybean Immature Seed using Fluorescent Derivative and LC/MS System.

    OHTAKE Norikuni, SATO Akiyo, SUEYOSHI Kuni, OHYAMA Takuji

    Plant and cell physiology   40   s52 - s52   1999年3月

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  • Comparison of phosphoenolpyruvate carboxylases in seeds of maize (Zea mays) and rice (Oryza sativa)

    Sasai Michitaka, Kamata Toru, Yamauchi Yasuo, Sueyoshi Kuni, Sugimoto Toshio, Oji Yoshikiyo

    神戸大学大学院自然科学研究科紀要. A   17   11 - 18   1999年3月

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    記述言語:英語   出版者・発行元:神戸大学  

    CiNii Article

    CiNii Books

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  • EFFECTS OF INHIBITORS ON EXPRESSION OF NITRATE REDUCTASE GENE IN BARLEY

    SUEYOSHI Kuni, MITSUYAMA Takashi, SUGIMOTO Toshio, OJI Yoshikiyo

    Plant and cell physiology   38   s62   1997年3月

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  • EXPRESSION OF ASPARAGINE SYNTHETASE IN GERMINATING RICE SEED

    KAWACHI Tahei, SUEYOSHI Kuni, SUGIMOTO Toshio, OJI Yoshikiyo

    Plant and cell physiology   38   s49   1997年3月

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  • 植物硝酸還元酵素のリン酸化,脱リン酸化による制御 解明進む環境条件に応答する巧妙な制御のしくみ

    末吉 邦, 杉本 敏男, 王子 善清

    化学と生物   34 ( 7 )   424 - 425   1996年7月

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  • Purification of Two Major Proteases in Cucumber Leaves

    YAMAUCHI Yasuo, ONISHI Michihito, SUGIMOTO Toshio, SUEYOSHI Kuni, OJI Yoshikiyo

    Plant and cell physiology   37   12 - 12   1996年3月

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  • FUNCTION OF PROTEASES EXPRESSING INCUCUMBER LEAF

    YAMAUCHI Yasuo, KIMURA Tomoko, SUEYOSHI Kuni, SUGIMOTO Toshio, OJI Yoshikiyo

    Plant and cell physiology   36   S78   1995年3月

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  • INTERCELLULAR FATE OF NITRATE REDUCTASE IN HIGHER-PLANTS - DEGRADATION AND INACTIVATION MECHANISMS

    H NAKAGAWA, K SUEYOSHI

    SEIKAGAKU   60 ( 3 )   206 - 211   1988年3月

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    記述言語:日本語   掲載種別:書評論文,書評,文献紹介等   出版者・発行元:JAPANESE BIOCHEMICAL SOC  

    Web of Science

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共同研究・競争的資金等の研究

  • オオムギ高親和性硝酸イオン輸送系の機能発現機構解明にむけた組織細胞学的アプローチ

    研究課題/領域番号:26450076

    2014年4月 - 2018年3月

    制度名:科学研究費助成事業

    研究種目:基盤研究(C)

    提供機関:日本学術振興会

    末吉 邦, 野澤 大樹, 高田 雅大

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    配分額:4940000円 ( 直接経費:3800000円 、 間接経費:1140000円 )

    本研究では、オオムギ根の高親和性硝酸輸送系を構成する2つのタンパク質(HvNRT2.1およびHvNAR2.3)の相互作用について、組織細胞生物学的に明らかにした。まず、2つのタンパク質は根の皮層における最も外側の細胞層の細胞膜に存在していることを明らかにした。続いて、HvNRT2.1とHvNAR2.3が細胞膜上で相互作用することをBiFC法で明らかにした。さらに、2つのタンパク質をタマネギ表皮細胞で個別に発現させたところ、それぞれのタンパク質単独では細胞膜に局在できないことを明らかにした。以上の結果より、両タンパク質が細胞膜に局在するためにはお互いの存在が必要であることが示唆された。

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  • ダイズにおける根粒固定窒素と根吸収窒素の移動機構と窒素固定硝酸阻害の網羅的解析

    研究課題/領域番号:26292036

    2014年4月 - 2017年3月

    制度名:科学研究費助成事業

    研究種目:基盤研究(B)

    提供機関:日本学術振興会

    大山 卓爾, 末吉 邦, 大竹 憲邦

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    配分額:16900000円 ( 直接経費:13000000円 、 間接経費:3900000円 )

    根粒菌を接種し無窒素水耕栽培したダイズに硝酸またはアンモニアを与えた後の根と根粒の代謝産物濃度の変動及びRNAseqによる遺伝子発現解析を実施した.窒素投与後、根および根粒でフルクトース及びグルコース濃度が減少し,アンモニウム処理により根で有機酸濃度の減少がみられた.
    硝酸供給8時間後の根,根粒の遺伝子発現をRNAseqにより解析した結果,特異的に発現が上昇した遺伝子数は根では12,939あり根粒では2,431あった。根と根粒で共通して上昇した遺伝子は74と少なかった.逆に硝酸投与で減少した遺伝子は,根では8,678, 根粒では1,654あり, 共通は42であった.

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  • オオムギ2成分高親和性硝酸イオン輸送系の機能解析

    研究課題/領域番号:21580076

    2009年 - 2011年

    制度名:科学研究費助成事業

    研究種目:基盤研究(C)

    提供機関:日本学術振興会

    末吉 邦

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    配分額:4810000円 ( 直接経費:3700000円 、 間接経費:1110000円 )

    オオムギ根は、高親和性硝酸輸送系(HATS)を介して低濃度の硝酸を吸収する。本研究では、HATSを構成する2つのタンパク質、HvNRT2およびHvNAR2の機能解明につながる知見を得た。まず、2つのタンパク質が細胞膜上で複合体を形成し得ることをin vitroで明らかにした。続いて、根の根毛域において、皮層の最も外側の細胞層に共に局在することを明らかにした。また、分割蛍光タンパク質を利用して、HvNRT2とHvNAR2が植物細胞の細胞膜上で相互作用して存在することをin vivoで明らかにした。

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  • オオムギ葉における低親和性硝酸イオントランスポーターの機能解明

    研究課題/領域番号:19580065

    2007年 - 2008年

    制度名:科学研究費助成事業

    研究種目:基盤研究(C)

    提供機関:日本学術振興会

    末吉 邦

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    配分額:4680000円 ( 直接経費:3600000円 、 間接経費:1080000円 )

    植物の無機窒素源である硝酸イオンは、根で吸収され、道管を介して葉に送られる。硝酸イオンの葉細胞内への輸送は、低親和性硝酸イオントランスポーター(NRT1)が関与すると予測されるが、その証拠はない。本研究では、オオムギで同定されたNRT1(HvNRT1)の機能解明を目的として研究を行い、(1)HvNRT1遺伝子は、地上部・地下部の両方で構成的に発現していること、(2)HvNRT1は細胞膜に発現していること、(3)HvNRT1を発現させたアフリカツメガエル卵母細胞は、低親和性硝酸イオン輸送活性を持つこと、を明らかにした。これらのことは、オオムギの葉細胞への硝酸イオン輸送にHvNRT1が関与している可能性を示している。

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  • ダイズ根粒における根粒形成自己制御と硝酸阻害の代謝調節機構の解析

    研究課題/領域番号:18380049

    2006年 - 2009年

    制度名:科学研究費助成事業

    研究種目:基盤研究(B)

    提供機関:日本学術振興会

    大山 卓爾, 末吉 邦, 大竹 憲邦

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    配分額:16570000円 ( 直接経費:15100000円 、 間接経費:1470000円 )

    ダイズの根粒生長と窒素固定活性の急速かつ可逆的な阻害が、アンモニア、尿素、グルタミンを培養液に投与した場合にも硝酸と同様に認められた。窒素化合物による根粒生長阻害は、^<13>Cと^<15>Nを用いた実験から光合成産物の根粒への移動低下が原因であると推定された。初生葉の葉柄から各種化合物を投与する実験で、グルタミンを投与したときに顕著な根粒形成の抑制が認められたことから、グルタミンが制御シグナルである可能性が示唆された。

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  • 植物の硝酸イオン輸送系タンパク質の翻訳後修飾に関する生化学的研究

    研究課題/領域番号:14360039

    2002年 - 2005年

    制度名:科学研究費助成事業

    研究種目:基盤研究(B)

    提供機関:日本学術振興会

    末吉 邦, 大山 卓爾, 大竹 憲邦

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    配分額:14600000円 ( 直接経費:14600000円 )

    植物において培地からのNO_3^-吸収に関わる硝酸トランスポーター(HvNRT1およびHvNRT2)とNO_3^-の道管ローディングに関わると予想されるアニオンチャネル(HvCLC)の翻訳後修飾に関する生化学的解析を加えることを目的に研究を行い,以下の結果を得た。
    1.オオムギ高親和性硝酸トランスポーター(HvNRT2)の翻訳後制御とタンパク質リン酸化
    オオムギを高濃度の窒素で生育させると、根の高親和性硝酸吸収活性は強く抑制されたが、根のNRT2タンパク質は高発現していた。これより、オオムギにおける高親和性硝酸吸収活性は、タンパク質の翻訳後レベルで制御されることが明らかとなった。組換えで得られたHvNRT2のC末端ポリペプチドは、オオムギ根の可溶性画分中のリン酸化酵素によりリン酸化された。このリン酸化活性は、Ca^<2+>依存的で、かつ窒素誘導性であった。オオムギ幼植物根より調製したミクロソーム画分中のHvNRT2タンパク質もまた可溶性画分によってリン酸化された。このリン酸化活性もCa^<2+>依存的で、かつ窒素誘導性であった。
    2.オオムギアニオンチャネル(HvCLC)のタンパク質リン酸化
    HvCLCタンパク質のC末端領域を大腸菌内で発現させ、組換えポリペプチドを得た。このHvCLCのC末端ポリペプチドは、オオムギ根の可溶性画分中のリン酸化酵素によりCa^<2+>に強く依存的してリン酸化された。このリン酸化活性は、培地窒素には応答しなかった。

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  • ダイズ根粒形成を制御する体内移動シグナル物質の検索

    研究課題/領域番号:13460030

    2001年 - 2003年

    制度名:科学研究費助成事業

    研究種目:基盤研究(B)

    提供機関:日本学術振興会

    大山 卓爾, 大竹 憲邦, 末吉 邦

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    配分額:15800000円 ( 直接経費:15800000円 )

    ダイズは、根に根粒を形成して空中窒素を固定利用することができる。一方、ダイズは根粒の過剰着生を抑制するための根粒形成自己制御(オートレギュレーション)機構を有することが知られている。近年、化学変異剤により、オートレギュレーション機能が欠損ないし、低下した根粒超着生変異株が得られた。根粒超着生形質は、地上部に支配されており、根粒菌の感染と根粒原基の形成により、「感染シグナル」が地上部へ伝達され、それを受け取った地上部(葉身)から地下部へ根粒の生長を抑制する「オートレギュレーションシグナル」が伝達されると予想されている。しかしながら、感染シグナルもオートレギュレーションシグナルも物質レベルでは同定されていない。本研究では、根粒超着生変異株と親株を用い、シグナル物質の探索を行った。
    シグナル物質が含まれると予想される篩管液の採取に成功した。篩管液と導管液中の成分をGC-MSで分析し、根粒菌接種後に変化する成分ピークの検索および、インドール酢酸とサリチル酸の分析を試みたが、これらの成分の検出には至らなかった。
    ダイズ品種Williamsとそれより分離された根粒超着生変異株NOD1-3のインドール酢酸とアブシジン酸をGC-MSで分析したところ、変異株根粒内のインドール酢酸濃度は親株の数分の1と低かった。地上部から輸送されるインドール酢酸がオートレギュレーションシグナルである可能性がある。
    病原菌の感染を全身に伝達するシグナル物質であるサリチル酸を、親株と変異株に投与したところ、親株では根粒形成の著しい抑制が見られたが、変異株では影響が少なかった。この結果はサリチル酸が根粒歯の感染シグナルである可能性を示唆する。
    根粒は、培地硝酸により可逆的かつ達やかに生長が阻害され、それは、光合成産物の分配が低下するためであることを発見した。NOD1-3は、硝酸存在下でも根粒への光合成産物の供給があまり低下せず、根粒の生長と窒素固定活性を維持した。

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  • 導管における硝酸イオンを介した器官間情報伝達の分子機構

    研究課題/領域番号:12025208

    2000年

    制度名:科学研究費助成事業

    研究種目:特定領域研究(A)

    提供機関:日本学術振興会

    末吉 邦, 大竹 憲邦, 大山 卓爾

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    配分額:2000000円 ( 直接経費:2000000円 )

    1.根柔細胞から導管への硝酸イオンのローディングに及ぼすアニオンチャネル阻害剤の影響について詳細に調べた.インタクトのオオムギ根にアニオンチャネル阻害剤(DIDS,A-9-C)を投与した後に硝酸塩を与え,導管液中の硝酸イオン濃度を調べた。R型アニオンチャネル阻害剤(DIDS)およびS型アニオンチャネル阻害剤(A-9-C)を用いたとき,硝酸イオンの導管への移行はほぼ50から70%抑制された。一方,根による培地からの硝酸イオンの吸収はいずれのアニオンチャネル阻害剤の影響を受けなかった.このことは,硝酸イオンは根で吸収された後,アニオンチャネルを介して導管へ放出されるものと考えられた.
    2.オオムギ根からアニオンチャネル遺伝子のクローニングとその発現解析を行った.
    他の植物や動物のアニオンチャネル遺伝子の塩基配列間で相同性の高い領域をRT-PCRで増幅し,この断片をプローブとしてオオムギ根cDNAライブラリーより全長cDNA(HvCLC)のクローニングを行った。その結果,2300bpのcDNAが得られた。塩基配列分析の結果,HvCLCはタバコおよびシロイヌナズナのアニオンチャネルcDNAと高い相同性を示した(約65%).このcDNAをプローブとしてノーザン解析をおこなったところ,HvCLCは培地の硝酸イオンの有無に関系なく一定量の発現がみられ,mRNA蓄積量は葉より根の方が多かった。また根の中心柱と皮層組織に同程度の発現がみられた.

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  • 硝酸還元酵素の翻訳後制御に関わる14-3-3タンパク質の細胞生物学的研究

    研究課題/領域番号:11660060

    1999年 - 2000年

    制度名:科学研究費助成事業

    研究種目:基盤研究(C)

    提供機関:日本学術振興会

    末吉 邦, 大山 卓爾

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    配分額:3500000円 ( 直接経費:3500000円 )

    植物を明所から暗所に移したときに起こるNRの急激な失活は、NRタンパク質のリン酸化と、それに続く14-3-3タンパク質の作用による。本研究では、14-3-3タンパク質の機能発現を細胞生物学的に解析した。
    1.免疫組織化学的解析のための特異抗体の精製
    タンパク質の免疫組織化学的解析には、高純度の特異抗体が必須となる。まず,大腸菌で発現させた組み換えオオムギ由来の14-3-3タンパク質を抗原としてウサギに免疫し抗血清を得た。次に,抗血清を14-3-3タンパク質を固定化したカラムに供試することによって,抗体を精製した。この抗体は非特異的に他のタンパク質を認識することの無いことが確認できた。
    2.発芽中の種子から胚乳と胚を分けて採取した。幼植物の地上部からは、展開中および未抽出の葉身、根からは皮層と中心柱を採取した。各部位の試料について、14-3-3タンパク質に対するウェスタンブロット分析を行ったところ、14-3-3タンパク質は全ての組織に同程度の含量で存在することが分かった。
    3.14-3-3タンパク質の免疫組織化学的解析
    葉および根における14-3-3タンパク質の組織内局在性の解析を試みた。葉組織切片は,固定包埋後,抗14-3-3タンパク質を一次抗体,FITCラベルの2次抗体として蛍光染色した。その結果,14-3-3タンパク質の存在を示唆する強いシグナルが維管束組織に観察された。根組織切片は,固定包埋後,抗14-3-3タンパク質抗体を反応させABC法で染色した。その結果,14-3-3タンパク質の存在を示唆する強いシグナルが,皮層細胞の最も外側と中心柱の内鞘細胞に認められた。

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  • 導管における硝酸イオンを介した器官間情報伝達の分子機構

    研究課題/領域番号:11151210

    1999年

    制度名:科学研究費助成事業

    研究種目:特定領域研究(A)

    提供機関:日本学術振興会

    末吉 邦, 中野 優, 大山 卓爾

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    配分額:2000000円 ( 直接経費:2000000円 )

    「目的」硝酸イオンが導管内を移動し、葉に到達する過程で、どのように硝酸同化系遺伝子の発現制御因子としての機能を発現するのかを明らかにすることを目的とする。本年度は、葉におけるNR遺伝子発現と導管内での硝酸イオンの輸送動態の関連性を詳細に解析することと、硝酸イオンの導管輸送坦体の性質について検討を加えた。
    「結果と考察」1.植物を無硝酸培地に移し硝酸イオンの供給を止めると、葉の細胞内に硝酸イオンが十分あるにもかかわらず、NRmRNA量と導管液硝酸イオン濃度は急速に低下した。このことより、葉におけるNR遺伝子の発現に関して、細胞内の硝酸イオンはシグナルとしての機能を持たず、導管液中の硝酸イオンがシグナルとして作用していると結論づけられた。2.オオムギ根にアニオンチャネルブロッカー(DIDS)を与えると、導管液中の硝酸イオン濃度が著しく減少した。一方、培地から根への硝酸イオンの吸収はDIDSで抑えられなかった。DIDSは、根の柔細胞から導管への硝酸イオンの積みこみの段階を阻害していると考えられた。すなわち、根の柔細胞から導管への硝酸イオンの積みこみ(loading)にアニオンチャネル様タンパク質の関与が考えられた。
    「今後の展望」1.今年度は、導管液中の硝酸イオンがNR遺伝子発現の誘導を引き起こすシグナルであることを示すことができた。導管は、葉の細胞にとっては細胞外空間である。今後は、細胞外シグナルとしての硝酸イオンを検知する仕組みを明らかにする必要がある。2.今年度は、導管への硝酸イオンのloadingにアニオンチャネルが関与する可能性を示すことができた。今後、チャネルの同定が必要である。

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  • 導管における硝酸イオンを介した器官間情報伝達の分子機構

    研究課題/領域番号:10170220

    1998年

    制度名:科学研究費助成事業

    研究種目:特定領域研究(A)

    提供機関:日本学術振興会

    末吉 邦, 王子 善清, 大山 卓爾

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    配分額:2500000円 ( 直接経費:2500000円 )

    「目的」本研究は、硝酸イオンが導管内を移動し、葉の細胞に供給される過程でどのようにシグナル機能を発現し、硝酸同化系遺伝子の発現を制御しているのかを明らかにすることを目的とする。具体的には、(1)導管以外に存在する硝酸イオンのシグナル機能の有無、(2)シグナルの受容機構、(3)導管中の硝酸イオン濃度を制御している因子の探索を目的とする.
    「経過」まず、葉におけるNR,NiR両遺伝子の発現レベルと導管以外のコンパートメントの硝酸イオン濃度に相関があるかを調べた.つまり、植物を無硝酸培地に移した後の葉におけるNR、NiR両mRNA蓄積量と葉部アポプラスト液および細胞質中の硝酸イオン濃度を経時的に測定し、変化を比較した。次に、葉と根における高親和性硝酸イオン輸送体遺伝子発現の硝酸イオン応答性を調べた.さらに、根の導管柔細胞より導管への硝酸イオンの積み込みに関わる遺伝子の探索を試みた.すなわち、オオムギ根の中心柱および皮層由来のcDNAを用いてcDNAサブトラクション法により中心柱特異的に発現している遺伝子をクローニングした.
    「結果・考察」植物を無硝酸培地に移し、硝酸の供給を止めるとNR、NiR両mRNA蓄積量と導管液硝酸イオン濃度は急速に低下したが、葉部アポプラスト液中の硝酸イオン濃度および細胞質硝酸イオン濃度の低下はわずかであった.以上のことより、葉における硝酸同化系遺伝子は、細胞質やアポプラスト中の硝酸イオンではなく、導管液中の硝酸イオンに応答して発現制御されていると結論づけた.ノーザン解析により、オオムギ高親和性硝酸イオン輸送体遺伝子は根にのみ発現していた。高親和性の硝酸イオン輸送体は葉における硝酸イオンの取り込みに関与していないことが示唆された.中心柱に特異的に発現している遺伝子60クローンを得た。全クローンの塩基配列分析を終了したが、硝酸イオンの導管への積み込みに関与する遺伝子はまだ得られていない。

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  • オオムギ硝酸還元酵素アイソザイムの異なる機能発現制御機構の解明

    研究課題/領域番号:09760057

    1997年 - 1998年

    制度名:科学研究費助成事業

    研究種目:奨励研究(A)

    提供機関:日本学術振興会

    末吉 邦

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    配分額:2100000円 ( 直接経費:2100000円 )

    「目的」
    オオムギには、活性に必要な電子供与体としてNADHのみを用いるNADH-NRと、NADHとNADPHの両方を用いるNAD(P)H-NRの2つのアイソザイムが存在する.しかし、硝酸同化における両者の機能的な差異は全く不明である.本研究では、硝酸イオン、代謝産物、光、ホルモンなどに対する両NRの機能発現応答の様式の違いを、転写レベル、タンパク質の発現レベルで詳細に解析することが目的である。
    「経過」
    前年度までに、硝酸イオンによるNRの発現誘導、同化窒素であるグルタミンやアスパラギンによる発現の抑制を調べ、両NR間で差異はないことを報告した.今年度は、オオムギ幼植物を明暗周期の条件下におき、NR活性およびNRタンパク質の変動を調べた.
    「結果と考察」
    オオムギを明暗周期の下で生育させ、明期および暗期開始後30分の植物より採取した葉をそれぞれ明処理葉および暗処理葉とした.暗処理葉から得た粗抽出液のNADH NRはMg^<2+>存在下で不活性化された.イムノブロット解析では、不活性化に伴ってNRタンパク質の減少はみられなかった.また、植物を暗所から明所に戻すとNR活性は元のレベルまで戻った.従って、NADHNRは暗条件に応答して可逆的に不活性化されると結論づけた.NAD(P)HNR活性は、野生型オオムギ葉において微弱であったため同様の解析が困難であった.NAD(P)HNR活性が葉において高く発現している変異株の種子を入手したので、これを用いて今後同様の解析を行う.

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  • 植物における導管を介した無機窒素栄養物質の輸送とその制御

    研究課題/領域番号:09274218

    1997年

    制度名:科学研究費助成事業

    研究種目:重点領域研究

    提供機関:日本学術振興会

    末吉 邦, 杉本 敏男, 王子 善清

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    配分額:3500000円 ( 直接経費:3500000円 )

    「目的」本研究では、硝酸イオンが導管内を輸送される過程でどのようにシグナルとしての機能を発現し、葉での硝酸同化系遺伝子の発現を制御しているのかを明らかにすることを目的とした。
    「経過か」水耕栽培のオオムギ幼植物に硝酸塩を種々の濃度で与えたときの葉組織の硝酸イオン蓄積量、導管液および葉部アポプラスト液の硝酸イオン濃度および硝酸同化系遺伝子の転写産物量を経時的に測定した。また、切断葉に対しても同様な実験を行った。オオムギの根を中心柱と皮層に分け、それぞれから単離した全RNAより作成したcDNAを用いてcDNAサブトラクション法を行い中心柱特異的に発現していると期待される遺伝子をクローニングした。
    「結果と考察」培地に硝酸塩を低濃度(μM)あるいは高濃度(mMレベル)で与えても、導管液中の硝酸イオン濃度は速やかに20-30mMに達した。その植物を無硝酸培地に移すと導管液の硝酸イオン濃度は急速に低下した。葉部アポプラスト液の硝酸イオン濃度は、導管液の硝酸イオン濃度の変化に連動して変動したが、その濃度範囲は1-10mMの間であった。一方、葉での硝酸還元酵素(NR)および亜硝酸還元酵素(NiR)遺伝子の発現量は高いレベルで維持されていた。これは導管液の硝酸イオン濃度レベルが高濃度で一定に保たれているという結果と合致する。一方、切断葉に低濃度(μMレベル)の硝酸イオンを与えた場合はNR、NiR遺伝子の転写産物量は検出できなかった。このことは、葉での硝酸同化系遺伝子の高い発現量を維持するため、導管における硝酸イオン濃度が数mM以上に保たれる必要があることを示す。中心柱全RNA由来のcDNAをtester、皮層全RNA由来のcDNAをdriverとしてサブトラクション法を行い、現在までに75クローンを得た。そのうちの5個については、ノーザン解析で中心柱特異的発現が確認できた。

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  • 登熟期に与える窒素肥料による種子内物質代謝の変動の解析と品質制御

    研究課題/領域番号:07660083

    1995年 - 1996年

    制度名:科学研究費助成事業

    研究種目:基盤研究(C)

    提供機関:日本学術振興会

    杉本 敏男, 末吉 邦, 王子 善清

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    配分額:2200000円 ( 直接経費:2200000円 )

    [目的]ダイズ種子のタンパク質の脂質の含量は品種や栽培環境により変動するが、どの様な機構によるか不明である。そこで、開花期に窒素肥料を与え、種子の成分含量への影響を調べた。[方法]2品種のダイズ(高蛋白品種:エンレイ“EN"、高脂質品種:タマホマレ“TM")を95年6月1日に播種し6月中旬に圃場に1株当たり2個体(株間隔20cm、1処理区5x1m、2連)を定植した。開花が始まってからしばらくの8月1日に1株当たり10gの硫安を株元にまいた(窒素施肥区、HN)。開花開始から60日頃から10日間隔で登熟途中の種子を3回採取し、凍結保存した。完熟種子は植物体が枯れた後に回収した。登熟途中と完熟した種子を重量の違いにより分離し、各々のタンパク質含量,脂質含量、アミノ酸と糖組成を調べた。[結果]両品種ともに窒素追肥の有無による種子の生長は、僅かに無施肥の方が速かった。タンパク質含量はENでは無追肥区(LN)で42〜44%、NHで40〜42%、TMではLNで36.5〜37.5%、HNで34〜36.5%で、種子重がENで0.23g,TMで0.24gで最も低くなった。また、脂質含量は全く逆になり、ENではLNで13〜14.5%、HNで14.5〜16%、TMでそれぞれ15.5〜16%、16〜16.5%になった。登熟途中の変化を調べると、脂質は登熟初期での蓄積が速く、窒素施肥により増量した。一方、タンパク質はLNの方が速く蓄積した。遊離アミノ酸組成では、転流形態の主要なアミノ酸のグルタミン含量がHNで極端に少なくなった。これらのことから、窒素施肥により茎葉部の生育が盛んになり光合成が盛んになったことに依る糖の種子への転流量が増加する一方で、アミノ酸の転流が抑制され、その結果として、種子の成分含量に影響が出たと考えられた。

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  • オオムギ硝酸還元酵素メッセンジャーRNAの安定性の制御機構に関する研究

    研究課題/領域番号:07760063

    1995年

    制度名:科学研究費助成事業

    研究種目:奨励研究(A)

    提供機関:日本学術振興会

    末吉 邦

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    配分額:1000000円 ( 直接経費:1000000円 )

    高等植物の硝酸還元酵素(NR)遺伝子は、培地での硝酸塩存在下でのみ発現する。本研究は、硝酸塩を与えて育てたオオムギ幼植物を無窒素培地に移したときのNRmRNAの急激な現象が、硝酸塩除去によって引き起こされたNRmRNAの不安定化によるものか、NRmRNAのもつ代謝回転速度が元来短いのかを評価しようと言うものである。
    オオムギ幼植物の切断葉を硝酸塩を含む培養液に4時間おいてNR遺伝子の発現を誘導した後、RNA合成阻害剤(コルジセピン)を含む培養液(硝酸塩は共存したまま)、あるいは無窒素培養液に移して経時的に試料を採取した。試料より全RNAを精製し、アガロースゲル電気泳動で分離後、ナイロン膜に転写し、^<32>PでラベルしたNRcDNAとハイブリダイズさせた。ナイロン膜に結合したプローブの放射能量をイメージングアナライザーで測定し、NRmRNA減少のキネティックスを調べた。
    硝酸塩の除去に伴うNRmRNAの半減期は、およそ30分であった。一方コルジセピンによってRNA合成を停止させた場合のNRmRNAの半減期は40分と測定された。この結果から、硝酸塩除去によって観察されるNRmRNAの急速な減少は、mRNAの不安定化が誘起されたからとは考えにくく、NRmRNAがもともと短い代謝回転速度を持っていることを示している。すなわち、NR遺伝子の転写は培地からの硝酸塩の除去によって停止させられ、細胞内に残存するNRmRNAが短時間で分解されていることを意味する。本研究の結果、NRmRNAの代謝回転速度が短いものであることが初めて示された。現在、植物は硝酸塩というシグナルの有無だけでNR遺伝子の発現を制御しうるのか検討しているところである。

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  • 硝酸還元酵素のリン酸化・脱リン酸化に対する阻害剤の影響

    研究課題/領域番号:06660079

    1994年

    制度名:科学研究費助成事業

    研究種目:一般研究(C)

    提供機関:日本学術振興会

    王子 善清, 末吉 邦, 杉本 敏男

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    配分額:1600000円 ( 直接経費:1600000円 )

    高等植物の硝酸還元酵素(NR)は光(明暗)環境の変化に伴ってリン酸化・脱リン酸化を受け、その活性を微妙に調節していることを確証するため、コマツナ葉を供試してin vivo及びin vitroでプロテインキナーゼとプロテインホスファターゼの阻害剤を活用して検証を行った。まず、in vivoの実験で、暗処理によるNRのマグネシウム感受性の増大、再度の明処理によるその消失を確認した。ついで、明処理葉を遮光したとき、また暗処理葉を光量を変えて照射したときには遮光及び照射強度に応じてNRのMg感受性が変わることを確認した。暗処理によるNRのMg感受性の増大はリン酸化の阻害剤であるマンノースを予め与えておくと観察されないし、また脱リン酸化の阻害剤であるカリキュリンAを予め与えた暗処理葉を明所に移したときにはMg感受性の消失が認められなかった。次いでin vitroの実験で、葉抽出液をATPとMgの存在下で20度でインキュベイトするとNRのMg感受性が増大し、この増大をリン酸化の阻害剤(K-252a)が阻害した。-^<32>P ATPとインキュベイトすると、-^<32>PがNR分子に取り込まれ、この取り込みをK-252aが阻害した。さらに、暗処理葉抽出液を30度でインキュベイトするとMg感受性が減少し、この減少をカリキュリンAが阻害した。
    以上によって、コマツナNRは暗所でリン酸化され不活性型になり、明所で脱リン酸化され活性型に戻ることが判明した。すなわち硝酸同化の鍵酵素であるNRは明暗環境の変化に対応してリン酸化・脱リン酸化されることによってその活性が微調整されていることがわかった。

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    2018年
    機関名:新潟大学

  • 土壌植物資源科学演習 Ⅰ

    2012年
    -
    2016年
    機関名:新潟大学

  • 水を巡る農の旅

    2012年
    -
    2016年
    機関名:新潟大学

  • 機器分析化学 Ⅰ

    2012年
    -
    2016年
    機関名:新潟大学

  • 生命・食料科学博士特定研究Ⅰ

    2012年
    -
    2015年
    機関名:新潟大学

  • 生命・食料科学博士セミナーⅠ

    2012年
    -
    2015年
    機関名:新潟大学

  • 生命・食料科学特定研究BⅠ

    2012年
    -
    2013年
    機関名:新潟大学

  • 生命・食料科学セミナーBⅠ

    2012年
    -
    2013年
    機関名:新潟大学

  • 文献詳読Ⅰ

    2012年
    -
    2013年
    機関名:新潟大学

  • 土壌植物資源科学演習Ⅰ

    2011年
    -
    2019年
    機関名:新潟大学

  • 土壌植物資源科学演習Ⅱ

    2011年
    -
    2018年
    機関名:新潟大学

  • スタディ・スキルズA2

    2011年
    機関名:新潟大学

  • 自然科学総論Ⅳ

    2011年
    機関名:新潟大学

  • 土壌植物資源科学実験

    2010年
    -
    2018年
    機関名:新潟大学

  • 機器分析化学Ⅰ

    2009年
    -
    2018年
    機関名:新潟大学

  • 生体物質構造解析学

    2008年
    -
    2010年
    機関名:新潟大学

  • 植物栄養生理化学

    2007年
    -
    2020年
    機関名:新潟大学

  • 植物生理学

    2007年
    -
    2017年
    機関名:新潟大学

  • 化学実験

    2007年
    -
    2017年
    機関名:新潟大学

  • 化学

    2007年
    -
    2015年
    機関名:新潟大学

  • 生命と環境の化学 II

    2007年
    -
    2015年
    機関名:新潟大学

  • 植物環境生理化学

    2007年
    -
    2011年
    機関名:新潟大学

  • 機器分析化学

    2007年
    -
    2010年
    機関名:新潟大学

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