研究者詳細 - 最相　大輔

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酸化ストレスとミトコンドリア機能．

目黒直樹, 最相大輔, 中園幹生

蛋白質核酸酵素 50 ( 14 ) 2005年

Spontaneous brassinolide-insensitive barley mutant ‘uzu’ adapted to East Asia

Saisho, D, K. Tanno, M. Chono, I. Honda, H. Kitano, K. Takeda

Breed. Sci. 54 ( 4 ) 409 - 416 2004年12月

「イネ科作物の半わい性形質の機構と利用」-”緑の革命”と渦性オオムギとの比較-

最相大輔

資源生物科学シンポジウム 21st 2004年

A semidwarf phenotype of barley uzu results from a nucleotide substitution in the gene encoding a putative brassinosteroid receptor. 国際誌

Makiko Chono, Ichiro Honda, Haruko Zeniya, Koichi Yoneyama, Daisuke Saisho, Kazuyoshi Takeda, Suguru Takatsuto, Tsuguhiro Hoshino, Yoshiaki Watanabe

Plant physiology 133 ( 3 ) 1209 - 19 2003年11月

記述言語：英語

Brassinosteroids (BRs) play important roles throughout plant growth and development. Despite the importance of clarifying the mechanism of BR-related growth regulation in cereal crops, BR-related cereal mutants have been identified only in rice (Oryza sativa). We previously found that semidwarf barley (Hordeum vulgare) accessions carrying the "uzu" gene, called "uzu" barley in Japan, are non-responding for brassinolide (BL). We then performed chemical and molecular analyses to clarify the mechanisms of uzu dwarfism using isogenic line pairs of uzu gene. The response of the uzu line to BL was significantly lower than that of its corresponding normal line. Measurement of BRs showed that the uzu line accumulates BRs, similar to known BR-insensitive mutants. The marker synteny of rice and barley chromosomes suggests that the uzu gene may be homologous to rice D61, a rice homolog of Arabidopsis BR-insensitive 1 (BRI1), encoding a BR-receptor protein. A barley homolog of BRI1, HvBRI1, was isolated by using degenerate primers. A comparison of HvBRI1 sequences in uzu and normal barley varieties showed that the uzu phenotype is correlated with a single nucleotide substitution. This substitution results in an amino acid change at a highly conserved residue in the kinase domain of the BR-receptor protein. These results may indicate that uzu dwarfism is caused by the missense mutation in HvBRI1. The uzu gene is being introduced into all hull-less barley cultivars in Japan as an effective dwarf gene for practical use, and this is the first report about an agronomically important mutation related to BRs.

DOI： 10.1104/pp.103.026195

PubMed

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14. 半矮性オオムギ"渦"は農業上重要なブラシノステロイド関連変異体である

蝶野真喜子, 本多一郎, 銭谷晴子, 米山弘一, 最相大輔, 武田和義, 高津戸秀, 星野次汪, 渡邊好昭

植物化学調節学会研究発表記録集 38th ( 38 ) 31 - 31 2003年10月

記述言語：日本語出版者・発行元：植物化学調節学会

Brassinosterois (BRs) play important roles throughout plant growth and development. Despite the importance of clarifying the mechanism of BR-related growth regulation in cereal crops, BR-related cereal mutants have been identified only in rice. We previously found that semi-dwarf barley accessions carrying the "uzu" gene, called "uzu" barley in Japan, are non-responding for brassinolide (BL). We then performed chemical and molecular analyses to clarify the mechanisms of uzu dwarfism using isogenic line pairs of uzu gene. The response of the uzu line to BL was significantly lower than that of its corresponding normal line. Measurement of BRs showed that the uzu line accumulates BRs, similar to known BR-insensitive mutants. The marker synteny of rice and barley chromosomes suggests that the uzu gene may be homologous to rice D61,a rice homolog of Arabidopsis BR-insensitive 1 (BRI1), encoding a BR-receptor protein. A barley homolog of BRI1,HvBRI1,was isolated by using degenerate primers. A comparison of HvBRI1 sequences in uzu and normal barley varieties showed that the uzu phenotype is correlated with a single nucleotide substitution. This substitution results in an amino acid change at a highly conserved residue in the kinase domain of the BR-receptor protein. These results may indicate that uzu dwarfism is caused by the missense mutation in HvBRI1. The uzu gene is being introduced into all hull-less barley cultivars in Japan as an effective dwarf gene for practical use, and this is the first report about an agronomically important mutation related to BRs.

DOI： 10.18978/jscrpanb.38.0_31

渦遺伝子がオオムギ未熟胚由来カルスからの植物体再分化に及ぼす影響

力石和英, 最相大輔, 前川雅彦, 武田和義

育種学研究 2003年

オオムギ半矮性遺伝子‘渦’はブラシノステロイド非感受性変異に起因する．

最相大輔, 蝶野真喜子, 本多一郎, 渡邊好昭, 丹野研一, 北野英己, 武田和義

日本植物生理学会年会およびシンポジウム講演要旨集 44 S71 - S71 2003年

記述言語：英語掲載種別：研究発表ペーパー・要旨（国際会議）出版者・発行元：日本植物生理学会

オオムギの渦性は，3H染色体上に座乗する単一劣性遺伝様式を示す遺伝子uzuによって支配される半矮性形質を示す．渦性系統は耐倒伏性に優れた草型を示し、多肥密植条件で多収となる．同様の草型を呈し，「緑の革命」として知られるイネ及びコムギの半矮性形質は，何れもジベレリンに関わる変異によってもたらされているが，オオムギ渦性は古くからジベレリンとは無関係の変異であることが知られており，その遺伝的な制御機構については不明であった．本研究では，uzu遺伝子の同質遺伝子系統対を用いて形態的搏Iな側面から詳細に解析を行った．その結果，渦性系統は高温生育条件下では第一節間の伸長が抑制されるdm型の節間伸長様式を示すこと，ブラシノライド投与に対する応答性が低く，且つ内在性のブラシノステロイド類が過剰に蓄積していることが明らかとなった．これらの形質は，近年イネで報告されたブラシノステロイド非感受性変異体d61と酷似していた．そこで，並性（正常型）と渦性オオムギからD61相同遺伝子を単離し，その塩基配列を比較したところ，C末端側の機能性領域の中にアミノ酸変異につながる一塩基置換を見出した．この変異は，遺伝地図上で渦性と同座に位置付けられ，皮裸性や播性程度の異なる複数の渦性系統が同一の変異を有することを見出した．従って，オオムギ渦性はブラシノステロイドレセプターの変異に起因すると結論した．

DOI： 10.14841/jspp.2003.0.185.0

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塩ストレス条件下における塩抵抗性オオムギ根cDNAのEXPRESSED SEQUENCE TAG(EST)解析

杉本学, 矢野健太郎, 南角奈美, 最相大輔, 佐藤和広, 武田和義

日本分子生物学会年会プログラム・講演要旨集 26th 2003年

オオムギ半矮性遺伝子uzu座に連鎖するマイクロサテライトマーカーの遺伝的多様性

丹野研一, 南角奈美, 最相大輔, 武田和義

育種学研究 = Breeding research 4 399 - 399 2002年8月

醸造用オオムギ"はるな二条"のBACライブラリー構築

最相大輔, 川崎信二, 佐藤和広, 武田和義

育種学研究 = Breeding research 4 269 - 269 2002年8月

オオムギにおける半矮性「渦」の特性解析

武田和義, 最相大輔, 力石和英, 北野英巳

育種学研究 4 356 2002年

The gene for alternative oxidase-2 (AOX2) from Arabidopsis thaliana consists of five exons unlike other AOX genes and is transcribed at an early stage during germination

D Saisho, M Nakazono, KH Lee, N Tsutsumi, S Akita, A Hirai

GENES & GENETIC SYSTEMS 76 ( 2 ) 89 - 97 2001年4月

記述言語：英語出版者・発行元：GENETICS SOC JAPAN

We investigated the expressions of genes for alternative oxidase (AOX1a, AOX1b, AOX1c and AOX2) and genes for cytochrome c oxidase (COX5b and COX6b) during germination of Arabidopsis thaliana, and examined oxygen uptakes of the alternative respiration and the cytochrome respiration in imbibed Arabidopsis seeds. A Northern blot analysis showed that AOX2 mRNA has already accumulated in dry seeds and subsequently decreased, whereas accumulation of AOX1a mRNA was less abundant from 0 hours to 48 hours after imbibition and then increased. The increase of the capacity of the alternative pathway appeared to be dependent on the expressions of both AOX2 and AOX1a. On the other hand, steady-state mRNA levels of COX5b and COX6b were gradually increased during germination, and the capacity of the cytochrome pathway was correlated with the increase of expressions of the COX genes. Antimycin A, the respiratory inhibitor, strongly increased the expression of AOX1a but had no effect on the expression of AOX2. A 5'RACE analysis showed that AOX2 consists of five exons, which is different from the case of most AOX genes identified so far. Analysis of subcellular localization of AOX2 using green fluorescent protein indicated that the AOX2 protein is imported into the mitochondria.

DOI： 10.1266/ggs.76.89

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ATP synthesis inhibitors as well as respiratory inhibitors increase steady-state level of alternative oxidase mRNA in Arabidopsis thaliana

D Saisho, M Nakazono, N Tsutsumi, A Hirai

JOURNAL OF PLANT PHYSIOLOGY 158 ( 2 ) 241 - 245 2001年2月

記述言語：英語出版者・発行元：URBAN & FISCHER VERLAG

The oxygen uptake in mitochondria can be uncoupled from ATP synthesis by the product of a single gene, the alternative oxidase (AOX) gene, which may be involved in signal transduction from mitochondria to the nucleus. To better understand induction of this gene in higher plants, we investigated the effects of the cytochrome respiratory inhibitors and ATP synthesis inhibitors on the expression of the AOXs of Arabidopsis thaliana. A Northern blot analysis showed the AOX1a transcripts were increased by treatment of antimycin A and myxothiazol, which act on complex III of the respiratory chain, and by NaN3, which acts on complex IV. AOX1a mRNA was also strongly induced by oligomycin (F1F0-ATP synthase inhibitor) and weakly induced by 2,4-DNP (ATP synthesis uncoupler) at the maximum dose. These results indicate that not only inhibition of electron transfer but also inhibition of ATP synthesis in mitochondria induces the transcription of AOX1a.

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Expression of a gene encoding mitochondrial aldehyde dehydrogenase in rice increases under submerged conditions

M Nakazono, H Tsuji, YH Li, D Saisho, S Arimura, N Tsutsumi, A Hirai

PLANT PHYSIOLOGY 124 ( 2 ) 587 - 598 2000年10月

記述言語：英語出版者・発行元：AMER SOC PLANT PHYSIOLOGISTS

It is known that alcoholic fermentation is important for survival of plants under anaerobic conditions. Acetaldehyde, one of the intermediates of alcoholic fermentation, is not only reduced by alcohol dehydrogenase but also can be oxidized by aldehyde dehydrogenase (ALDH). To determine whether ALDH plays a role in anaerobic metabolism in rice (Oryza sativa L. cv Nipponbare), we characterized a cDNA clone encoding mitochondrial ALDH from rice (Aldh2a). Analysis of sub-cellular localization of ALDH2a protein using green fluorescent protein and an in vitro ALDH assay using protein extracts from Escherichia coli cells that overexpressed ALDH2a indicated that ALDH2a functions in the oxidation of acetaldehyde in mitochondria. A Southern-blot analysis indicated that mitochondrial ALDH is encoded by at least two genes in rice. We found that the Aldh2a mRNA was present at high levels in leaves of dark-grown seedlings, mature leaf sheaths, and panicles. It is interesting that expression of the rice Aldh2a gene, unlike the expression of the tobacco (Nicotiana tabacum) Aldh2a gene, was induced in rice seedlings by submergence. Experiments with ruthenium red, which is a blocker of Ca2+ fluxes in rice as well as maize (Zea mays), suggest that the induction of expression of Adh1 and Pdc1 by low oxygen stress is regulated by elevation of the cytosolic Ca2+ level. However, the induction of Aldh2a gene expression may not be controlled by the cytosolic Ca2+ level elevation. A possible involvement of ALDH2a in the submergence tolerance of rice is discussed.

DOI： 10.1104/pp.124.2.587

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オオムギのcDNAシークエンス解析 : ビールオオムギ「はるな二条」と日本在来品種「赤神力」との比較

最相大輔, 佐藤和広, 高橋秀和, 武田和義

育種学研究 = Breeding research 2 ( 2 ) 32 - 32 2000年9月

Transcript levels of the nuclear-encoded respiratory genes in rice decrease by oxygen deprivation: Evidence for involvement of calcium in expression of the alternative oxidase 1a gene

Hiroyuki Tsuji, Mikio Nakazono, Daisuke Saisho, Nobuhiro Tsutsumi, Atsushi Hirai

FEBS Letters 471 ( 2-3 ) 201 - 204 2000年4月

記述言語：英語

We investigated the effect of oxygen on the expressions of respiratory genes encoded in the nuclear and mitochondrial genomes of rice (Oryza sativa L.). Hypoxic treatment decreased the transcript levels of nuclear-encoded, but not mitochondrial-encoded respiratory genes. The effects of ruthenium red (an inhibitor of Ca2+ fluxes from organelles) and/or CaCl2 on plants under hypoxic conditions suggested that Ca2+ is a physiological transducer of a low-oxygen signaling pathway for expression of the alternative oxidase 1a gene (AOX1a), but not for expressions of genes involved in the cytochrome respiratory pathway, in rice. Copyright (C) 2000 Federation of European Biochemical Societies.

DOI： 10.1016/S0014-5793(00)01411-3

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Transcript levels of the nuclear-encoded respiratory genes in rice decrease by oxygen deprivation : evidence for involvement of calcium in expression of the alternative oxidase 1a gene.(共著)

FEBS Lett. 471 ( 2-3 ) 201 - 204 2000年4月

Transcript levels of the nuclear-encoded respiratory genes in rice decrease by oxygen deprivation: evidence for involvement of calcium in expression of the alternative oxidase la gene

H Tsuji, M Nakazono, D Saisho, N Tsutsumi, A Hirai

FEBS LETTERS 471 ( 2-3 ) 201 - 204 2000年4月

記述言語：英語出版者・発行元：ELSEVIER SCIENCE BV

We investigated the effect of oxygen on the expressionse of respiratory genes encoded in the nuclear and mitochondrial genomes of rice (Oryza sativa L.). Hypoxic treatment decreased the transcript levels of nuclear-encoded, but not mitochondrial-encoded respiratory gents. The effects of ruthenium red (an inhibitor of Ca2+ fluxes from organelles) and/or CaCl2 on plants under hypoxic conditions suggested that Ca2+ is a physiological transducer of a low-oxygen signaling pathway for expression of the alternative oxidase 1a gene (AOX1a), but not for expressions of genes involved in the cytochrome respiratory pathway, in rice. (C) 2000 Federation of European Biochemical Societies.

DOI： 10.1016/S0014-5793(00)01411-3

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低酸素によるイネAOX1a遺伝子の発現抑制には細胞内カルシウムイオンが関与している

辻寛之, 中園幹生, 最相大輔, 堤伸浩, 平井篤志

育種学研究 = Breeding research 2 ( 1 ) 66 - 66 2000年4月

Expression of a gene encoding mitochondrial aldehyde dehydrogenase in rice increases under submerged conditions.(共著)

Plant Physiol. 124 ( 2 ) 587 - 598 2000年

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