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        <FirstName EmptyYN="N">Masakazu</FirstName>
        <LastName>Hirotsu</LastName>
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    <ArticleTitle>追尾機能と電磁石スペクトロメータを備えた粒子望遠鏡の作製とシグナスX-3由来のミューオン観測</ArticleTitle>
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    <ArticleTitle>Mechanism for Deletion Formation Depending on Direct Repeats between Homeologous Sequences and between Nonhomologous Sequences:Involvement of the Escherichia coli Single- Stranded DNA-Binding Protein(SSB)</ArticleTitle>
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    <ArticleTitle>Self-Dual Codes, Designs and Unimodular Lattices</ArticleTitle>
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    <ArticleTitle>Studies of spontaneous and evoked neurotransmitter release at verte- brate and invertebrate neuromuscular junctions from the viewpoint of the mechanism for release</ArticleTitle>
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      <JournalTitle>Acta Medica Okayama</JournalTitle>
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        <Year>1997</Year>
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    <ArticleTitle>Transcriptional Regulation and DNA- Protein Interaction in Gene Expression in Arabidopsis thaliana</ArticleTitle>
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      <JournalTitle>Acta Medica Okayama</JournalTitle>
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        <Year>1997</Year>
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    <ArticleTitle>Cloning and Characterization of Flagellar Operons of the Four Shigella Subgroups:Genetic Defects that caused Flagella Loss of the Subgroups</ArticleTitle>
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      <PublisherName/>
      <JournalTitle>Acta Medica Okayama</JournalTitle>
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      <Volume/>
      <Issue/>
      <PubDate PubStatus="ppublish">
        <Year>1997</Year>
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    <ArticleTitle>薬物の抱合代謝体の体内動態特性の解析と薬物 代謝体間相互作用に関する研究</ArticleTitle>
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      <PublisherName/>
      <JournalTitle>Acta Medica Okayama</JournalTitle>
      <Issn/>
      <Volume/>
      <Issue/>
      <PubDate PubStatus="ppublish">
        <Year>1997</Year>
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    <ArticleTitle>抗悪性腫瘍薬ジノスタチン スチマラマーの作用とその機構に関する研究</ArticleTitle>
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  <Article>
    <Journal>
      <PublisherName/>
      <JournalTitle>Acta Medica Okayama</JournalTitle>
      <Issn/>
      <Volume/>
      <Issue/>
      <PubDate PubStatus="ppublish">
        <Year>1997</Year>
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    <ArticleTitle>トウダイグサ科植物に特有な加水分解性タンニンの化学的研究</ArticleTitle>
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  <Article>
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      <PublisherName/>
      <JournalTitle>Acta Medica Okayama</JournalTitle>
      <Issn/>
      <Volume/>
      <Issue/>
      <PubDate PubStatus="ppublish">
        <Year>1997</Year>
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    <ArticleTitle>Acinetobacter 属菌における1,3- diaminopropane生合成酵素の遺伝子解析に関する研究</ArticleTitle>
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      <PublisherName/>
      <JournalTitle>Acta Medica Okayama</JournalTitle>
      <Issn/>
      <Volume/>
      <Issue/>
      <PubDate PubStatus="ppublish">
        <Year>1997</Year>
        <Month/>
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    <ArticleTitle>ヒト型トロンボポエチンの血小板減少改善効果に関する研究</ArticleTitle>
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  <Article>
    <Journal>
      <PublisherName/>
      <JournalTitle>Acta Medica Okayama</JournalTitle>
      <Issn/>
      <Volume/>
      <Issue/>
      <PubDate PubStatus="ppublish">
        <Year>1997</Year>
        <Month/>
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    <ArticleTitle>脳由来神経栄養因子BDNF遺伝子の細胞内カルシウム流入による発現誘導機構の解析</ArticleTitle>
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    <Abstract/>
    <CoiStatement>No potential conflict of interest relevant to this article was reported.</CoiStatement>
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  <Article>
    <Journal>
      <PublisherName/>
      <JournalTitle>Acta Medica Okayama</JournalTitle>
      <Issn/>
      <Volume/>
      <Issue/>
      <PubDate PubStatus="ppublish">
        <Year>1997</Year>
        <Month/>
      </PubDate>
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    <ArticleTitle>アデノシン誘導体の体内動態の薬物速度論的解析</ArticleTitle>
    <FirstPage LZero="delete"/>
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    <Language>EN</Language>
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    <Abstract/>
    <CoiStatement>No potential conflict of interest relevant to this article was reported.</CoiStatement>
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  <Article>
    <Journal>
      <PublisherName/>
      <JournalTitle>Acta Medica Okayama</JournalTitle>
      <Issn/>
      <Volume/>
      <Issue/>
      <PubDate PubStatus="ppublish">
        <Year>1997</Year>
        <Month/>
      </PubDate>
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    <ArticleTitle>大腸菌のNa+ / H+ アンチポーター(NhaA,NhaB,ChaA)の機能と役割</ArticleTitle>
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    <Abstract/>
    <CoiStatement>No potential conflict of interest relevant to this article was reported.</CoiStatement>
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  <Article>
    <Journal>
      <PublisherName/>
      <JournalTitle>Acta Medica Okayama</JournalTitle>
      <Issn/>
      <Volume/>
      <Issue/>
      <PubDate PubStatus="ppublish">
        <Year>1997</Year>
        <Month/>
      </PubDate>
    </Journal>
    <ArticleTitle>Studies on Male Pronuclear Formation after In Vitro Fertilization of Pig Oocytes</ArticleTitle>
    <FirstPage LZero="delete"/>
    <LastPage/>
    <Language>EN</Language>
    <AuthorList>
      <Author>
        <FirstName EmptyYN="N">Ken</FirstName>
        <LastName>Sawai</LastName>
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    <Abstract>The objective of the present study was to examine the effects of various factors responsible for male pronuclear (MPN) formation associated with glutathione (GSH) synthesis of pig oocytes. In the first series of experiments, when cumulus-enclosed oocytes were cultured for 24 h in maturation medium supplenlented with 0.57 mM cysteine, GSH concentration of oocytes was increased. However, the activation of oocytes after sperm penetration in vitro was inhibited and thereby MPN formation of the oocytes was not promoted. The MPN formation in activated oocytes after sperm penetration at any stages of maturation was promoted when cysteine was added to maturation medium. The results indicate that an increased concentration of GSH and intimate synchronization with oocyte activation are essential for MPN formation of pig oocytes. In the next series of experiments, it was shown that, when cysteine was removed from maturation medium at 36 h of culture, both the incidence of MPN formation after in vitro fertilization and GSH concentration of lmatured oocytes were lower than when cysteine was present during whole period of culture for 48 h. In contrast, the presence of cysteine in maturation medium only bc~tween 42 and 48 h of culture, when oocytes reached to the late metaphase-I to metaphase-II stage, could promote oocyte GSH synthesis and thereby MPN formation after in vitro fertilization. The results indicate that the presence of cysteine in maturation medium for 6 h of the last phase of maturation is essential for oocyte GSH synthesis enough to induce MPN formation with the same efficiency as in oocytes matured in the presence of cysteine from the start of culture. The effects of cystine, an oxidized form of cysteine, added to maturation medium at various times after culture on the MPN formation and GSH synthesis of cumulus-enclosed and cumulus-free oocytes were examined in the third series of experiments. When maturation medium was supplemented with 0.57 mM cystine, MPN formation and GSH synthesis of cumulus-enclosed oocytes were accelerated, but not in cumulus-denuded oocytes. On the other hand, cysteine (0.57 mM) could promote MPN formation and GSH synthesis in both cumulus-enclosed and cumulus-denuded oocytes. The results indicate that cystine is associated with increased GSH synthesis and MPN formation of pig oocytes and that the presence of cumulus cells is essential for the utilization of the cystine by oocytes. In contrast, cysteine is utilized directly by oocytes without cumulus cells for GSH synthesis and MPN formation. Taken together, it is concluded from these results that synchronization of oocyte activation and sperm penetration, and the addition of exogenous compounds such as cysteine or cystine to maturation medium are important factors for GSH synthesis of matured oocytes and MPN formation of oocytes following fertilization. The presence of cumulus cells associated with utilization of cystine by oocytes is also an important factor.</Abstract>
    <CoiStatement>No potential conflict of interest relevant to this article was reported.</CoiStatement>
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  <Article>
    <Journal>
      <PublisherName/>
      <JournalTitle>Acta Medica Okayama</JournalTitle>
      <Issn/>
      <Volume/>
      <Issue/>
      <PubDate PubStatus="ppublish">
        <Year>1997</Year>
        <Month/>
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    </Journal>
    <ArticleTitle>宿主特異性決定における植物細胞壁の役割〜エンドウ･エンドウ褐紋病菌を用いた解析〜</ArticleTitle>
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    <Language>EN</Language>
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    <Abstract>植物一病原菌間の特異性決定機構の解明は、植物病理学において最重要課題の1つである。このメカニズムの解明に向けて、エンドウ褐紋病菌と宿主、非宿主植物を用いて解析した。エンドウ褐紋病菌は柄胞子が発芽する際に抵抗性誘導因子(エリシター)と特異性決定を担う抵抗性抑制因子(サプレッサー)を生産する。サプレッサーはエリシターによって誘導される複数の防御応答を抑制(遅延)する。近年、サプレッサーの作用点の解析が進められ、宿主のATPase阻害にあることが判った。実際に、植物葉をサプレッサーで処理すると、宿主エンドウのATPaseのみが阻害されること、さらにはp型ATPaseの阻害剤であるオルトパナジン酸がエンドウの防御応答を抑制することが明らかとなった。しかしながら、数種の植物から調製した原形質膜画分のATPase活性は、植物種に関係なく阻害され、組織で見られた特異性は認めらなかった。これらの結果は、宿主特異性決定には植物細胞壁が重要な役割を担っていることを強く示唆する。そこで、本研究ではこれら病原菌シグナルの認識機構や植物一病原菌間の特異性決定における細胞壁の役割について解析した。褐紋病菌の宿主エンドウ、非宿主ササゲより細胞壁画分を調製後、NaClを用いて可溶化タンパク質を得た。11 種のマーカー酵素活性を測定したところ、原形質膜をはじめとする他のオルガネラの混入はほとんど認められなかった。細胞壁可溶化画分中には薬剤感受性、基質特異性、至適pH、二価イオン要求性等の諸性質が原形質膜ATPaseとは異なるATPase(NTPase)が存在することが判った。本細胞壁ATPaseは褐紋病菌のエリシターによって非特異的に活性化され、サプレッサーによって種特異的に制御された。さらにATPアガロースカラム、陰イオン交換カラムを用いて部分精製した細胞壁ATPaseもエリシタ一、サプレッサーに同様な応答性を持つことが明らかとなった。このことはエリシターの植物組織に対する非特異的作用、およびサプレッサーの植物の防御応答に対する種特異的な作用を反映しており、植物細胞壁が病原菌の認識と宿主特異性決定に重要な役割を果たすこと、さらには細胞壁ATPase自体かその近傍に病原菌シグナルの受容体が存在する可能性を強く示唆している。そこで0(2)(一)生成を指標に無傷エンドウ、ササゲ葉における防御応答について調べた。無傷葉における0(2)(一)生成はエリシターで5分以内に誘導されたが、サプレッサーによって種特異的に制御されること、また0(2)(ー)は非病原菌接種で生成が誘導されるが、病原菌接種では誘導されないことが判った。さらに、エンドウ、ササゲの細胞壁可溶化画分中にはNADH依存性の0(2)(一)生成活性が存在し、ATPaseとまったく同様に病原菌シグナルに応答性を持つ(エリシターで非特異的に活性化し、サプレッサーによって種特異的に制御される)ことが判った。 このことは無傷エンドウ、ササゲ葉における0(2)(一)生成と一致しており、エンドウ、ササゲ葉における0(2)(-)の生成に細胞壁が深く関与することが判った。また、O(2)(-)生成のみならず、エンドウ、ササゲの細胞壁可溶化画分中にはH(2)0(2)生成、パーオキシダーゼ、アスコルビン酸オキシダーゼ、リンゴ酸脱水素酵素活性が存在し、このうちリンゴ酸脱水素酵素活性を除く活性は、エリシタ一、サプレッサーによってATPase と同調的に制御されること、さらにパーオキシダーゼは細胞壁ATPaseと精製過程における挙動が一致することが判った。このことから、病原菌シグナルの認識後、細胞壁における酸化、還元状態が速やかに変化すること、また酸化、還元状態の変化に必要な酵素群は細胞壁中で複合体(装置)を形成していることが推察された。このように病原菌シグナルの第一義的な認識の場は細胞壁であるものと考えられるが、多様な細胞応答に結びっくためには細胞壁で認識された情報は細胞内へ伝達される必要がある。そこで、細胞壁と原形質膜間の情報伝達系の検索を目的とし、細胞外マトリクスと細胞骨格の連結に関わる膜貫通タンパク質であるインテグリン様タンパク質の有無を調べ、エンドウの防御応答における役割について解析した。細胞外マトリクスとインテグリンの結合に関与するアルギニンーグリシンーアスパラギン酸(RGD) の配列を含む合成ペプチドはエリシターとの同時処理では有意な影響は認められなかったものの、前処理した場合には前処理時間の長さに依存したピサチン蓄積阻害が認められた。一方、RGDペプチドは原形質膜および細胞壁ATPaseを阻害することはなく、褐紋病菌サプレッサーとは作用は異なった。エンドウ原形質膜中にはインテグリン様タンパク質が複数存在し、それらのタンパク質中には細胞壁タンパク質、細胞骨格系タンパク質と相互作用する分子が存在することが明らかとなった。以上の結果からインテグリン様タンパク質を介した細胞壁一原形質膜(細胞内)間の情報伝達系が存在する可能性が示唆された。以上のように植物の細胞壁は病原菌シグナルの第一次作用部位であり、病原菌認識、宿主特異性決定、あるいはその後の防御応答に重要な役割を持つ可能性が強く示唆された。一方、エリシタ一、サプレッサーの受容体が細胞壁に存在することが想定されるがその実態については明らかではない。エンドウ褐紋病菌の生産するエリシターについては1分子が精製され、Glcsβ1-6Maα1-6Manの三糖がセリンを介してタンパク質鎖とO-グリコシド結合した、分子量70,000〜140,000の高分子糖タンパク質であることが明らかとなっている(Matsubara and Kuroda 1986)。そこでエリシター活性の最小単位、さらにはエリシターの受容体を同定するためのプローブを検索することを目的とし、Glcβ1-6Manα1-6Manの三糖を単位とする9種の糖ペプチドを合成し、エンドウの防御応答に対する影響を調べた。エンドウの防御応答について調べた。9種の合成糖ペプ
チドはピサチン蓄積を誘導したが、ピサチン蓄積誘導活性は褐紋病菌エリシターと比較して非常に弱かった。しかしながら、供試した9種の合成糖ペプチドのうちNo.2〜9はエンドウ組織に局部的な抵抗化を誘導し、合成糖ペプチドNo.4〜9はエンドウ組織表層におけるスーパーオキシドアニオン生成を誘導した。さらに、合成糖ペプチドNo.4〜9はin vitroにおいて細胞壁画分ATPaseを活性化した。また、合成糖ペプチドによ
る局部抵抗化スーパーオキシドアニオン生成の誘導、細胞壁画分ATPaseの活性化は合成糖ペプチドの分子量に依存している傾向が伺えた。以上の結果から、今回供試した合成糖ペプチドのうち特にNo.4〜9はエンドウ組織表層における防御応答のエリシターとして作用することが判った。このことから合成糖ペプチドNo.4〜9はエンドウの初期防御応答を解析、およびエリシターの受容体同定の有用なモデルエリシターとなりえるものと考えられる。以上の結果を総合すると、植物細胞の最外層に位置する植物固有のオルガネラである細胞壁が病原菌の認識、宿主特異性決定の場であり、さらには病原菌認識後の防御応答、あるいは防御応答誘導のための第二次シグナル生成の場として重要な働きを担うものと考えられる。また、エンドウ褐紋病菌はサプレッサーを分泌することによりエンドウの細胞壁ATPaseやそれと同調的に制御される酵素群の活性を阻害することにより、細胞内へのシグナルの伝達を阻止(撹乱)し、防御応答の発現を抑制(回避)することによって感染を成立させているものと推察できる。</Abstract>
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    <Abstract>本研究は，低コスト化や省力化，そして環境への負荷の軽減を目指したLISA (Low Input Sustainable Agriculture = 低投入持続的農業) を実現するために，水田生態系に対する撹乱的働きかけを最小限にとどめた不耕起水田において，マメ科緑肥作物で覆うことにより，合理的かつ適切な雑草管理の方向性を追求したものである．これらの研究成果の概要は以下の通りである．I． レンゲ草生マルチを活用した不耕起水稲作における雑草の発生消長　まず我が国で古くから水田冬作に緑肥作物として栽培されてきたレンゲを草生マルチの候補種とした．不耕起水田にレンゲを草生マルチとして活用した水稲直播栽培を3年間継続したところ，栽培試験の初年度は，レンゲはよく繁茂し冬生雑草の発生も少なかった．そして湛水後の夏生雑草の発生は，前年の慣行栽培時に多数発生したコナギが消滅したほか他の草種の発生も少なく，水稲収量も470kg/10a以上を得た．またレンゲ草生マルチをレンゲ開花期の5月中旬に除去すると， 湛水後に一年生のカヤツリグサ科やアゼナ属を主とした水田夏生一年生雑草の密度が顕著に増加し，レンゲマルチによってこれらの雑草の発生を抑制できることが明確となった．しかし2年目以降，レンゲ群落の衰退とスズメノテッポウ，ノミノフスマなどの冬生雑草の増加が認められ，それに伴いヒエ属などの水田夏生雑草の発生数も増加して水稲収量は激減した．従って，レンゲ草生マルチによる雑草管理の問題点として，(1)イヌビエやヒメタイヌビエなどのノビエの
発生に対しては抑制効果が劣ること (2)圃場に湛水を開始する時期が遅くなるとレンゲが枯死し，マルチ効果が薄れること (3)全く除草を行わないと次年度以降の発生ポテンシャルとなる埋土種子集団の増加を抑えきれないこと，を明らかにした．II．異なる種子深度および潅水条件下における数種の植物マルチ資材がヒメタイヌビエの発生に及ぼす影響　ヒエ属のヒメタイヌビエ種子を用いて，レンゲ，へアリーベッチ， シロクローバおよびヨシのマルチ資材と，マルチ量，種子埋土深度，そして海水条件の4要国を組み合わせて発生数の比較を試みた．分散分析(ANOVA)や数量化理論I類による解析の結果，ヒメタイヌビエ同子が地表面lに存在する場合にはマルチの存在によって有意に発生が抑制され，その抑制の度合いはマルチの量が多いほど，そして深水になるほど強まることが明らかになった．従って，マルチ草種としては現存量の多い草種ほど雑草の発生が制御できると考えられた．III． 草生マルチの雑草抑制能力と湛水時期との関係　1．レンゲ以外のマルチ草種の活用の可能性を検討するために，強いアレロパシーも報告されているマメ科一年生緑肥作物のへアリーベッチと多年生のシロクローバを圃場に栽培し，雑草抑制力をレンゲと比較した．その結果湛水時まで旺盛な生育を続けるシロクローバや，現存量がレンゲの約2倍あり，密な群落を
形成するために光遮蔽効果が枯死後も維持できるへアリーベッチが，レンゲよりも冬生雑草および湛水後の雑草の発生を抑えた．そしてへアリーベッチマルチ区に移植した水稲は登熟歩合や千粒重が高まり，488.6kg/10aの収量が得られた．2．湛水下のマルチが分解する過程で植物の発芽および生育に及ぼす影響をレタス種子を用いて検討したところ，枯死群落より生存群議への湛水で，また植物部位別では葉部から得た水抽出液が顕著にレタスの発芽を阻害することが明らかになった．そしてレタスの発芽および初期生育阻害は，水抽出液のECと関係があり，有機溶媒を用いた水抽出液の分画操作の結果，有機酸が阻害作用に関与していることが推察された．このことから，マルチ草種がまだ旺盛に生育し，葉部の比率が高い開花期頃までに湛水を開始すれば，雑草の発芽や生育を抑制する可能性が示唆された．IV．雑草防除法，耕起法および作付け様式の異なる水田における埋土種子および雑草発生消長の比較　雑草の埋土種子量と圃場の管理様式との関係を調べたところ，不耕起水田では埋土種子の分布が表層部に偏り，また除草剤連用水田や丹念に手取り除草を続けている圃場では，雑草の発生ポテンシャルが少ないことを把握した．そして積極的に雑草防除を行わない不耕起水田では，ヒエ属を中心に埋土種子量が多くなった．従って，草生マルチの活用による雑草管理においても，次年度以降の発生ポテンシャルの増加を抑えなければ，マルチで抑制できない雑草が増加することが示唆された．以上の試験結果に基づき，@初期生育がなるべく速やかで， 他雑草との競合に有利であること，A現存量が多く密度の高い群落を形成し，光遮蔽効果を保つこと，B アレロパシー活性が高いこと，C湛水開始期まで生存していること，D水稲栽培期間中，マルチが残存すること，E連作が可能で，毎年安定して繁茂すること E 湛水による枯死以前に種子を生産し，自然更新すること，が不耕起水田の雑草管理に適した理想的な草生マルチの条件と考えられた．最後にこれらの結果をもとに，マルチ草種と水稲との競合も考慮しつつ，圃場への湛水が5月上旬以前に可能となる地域ではレンゲ，そして湛水開始時期が6月中旬以降となる地域では枯れ上がりが遅く乾田期間中のマルチ効果が維持でき
るへアリーベッチを草生マルチとして活用する雑草管理の試案を提示した．</Abstract>
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