{"created":"2021-03-01T06:34:13.873398+00:00","id":28421,"links":{},"metadata":{"_buckets":{"deposit":"030380d5-a503-4c78-910a-96f62fad480a"},"_deposit":{"id":"28421","owners":[],"pid":{"revision_id":0,"type":"depid","value":"28421"},"status":"published"},"_oai":{"id":"oai:niigata-u.repo.nii.ac.jp:00028421","sets":["453:456","485:872:1583:1602"]},"item_7_alternative_title_1":{"attribute_name":"その他のタイトル","attribute_value_mlt":[{"subitem_alternative_title":"根こぶ病抵抗性カブ（Brassica rapa L.）培養根へのFura-2の導入と細胞内Ca2+ 濃度の測定"}]},"item_7_biblio_info_6":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2007-08","bibliographicIssueDateType":"Issued"},"bibliographicIssueNumber":"1","bibliographicPageEnd":"66","bibliographicPageStart":"61","bibliographicVolumeNumber":"60","bibliographic_titles":[{"bibliographic_title":"新潟大学農学部研究報告"},{"bibliographic_title":"新潟大学農学部研究報告","bibliographic_titleLang":"en"}]}]},"item_7_description_4":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"Our previous study using liquid organ cultured adventitious roots from turnip (Brassica rapa L.) showed that Ca2+ is required for induction of defense responses in the cultured roots on the treatment with Plasmodiophora brassicae resting spores (Takahashi et al., 2006). To evaluate change in [Ca2+]cyt in cultured root cells on contact with the spores, acetoxymethyl ester derivative of Fura-2 (Fura-2/AM) was loaded into the roots. When ionophore A23187 was treated with Ca2+ simultaneously, the Fura-2 fluorescence ratio that represents relative [Ca2+]cyt increased promptly, showing that the Fura-2/AM system is suitable to evaluate [Ca2+]cyt change in cultured root cells in a second range. Applicability of this method for studies on Ca2+ fluctuation against various extracellular stimuli was supported by observations that treatment with mannitol or NaCl also immediately increased the Fura-2 ratio. When the Fura-2/AM loaded roots were treated with resting spores, no [Ca2+]cyt change was observed during 500 second but when treated with spores pre-incubated with germination-enhancing suspension (GES), a slight but reproducible increase in [Ca2+]cyt was observed. We conclude that although further analysis is needed, the Fura-2/AM system will contribute to revealing the Ca2+ involvement in clubroot-resistance response. 我々は以前の研究において、カブ（Brassica rapa L.）幼根から誘導した培養根を用い、根こぶ病菌Plasmodiophora brassicaeに対する初期抵抗反応の誘導にCa2+ が必要であることを示した（Takahashi et al., 2006）。本研究では、根こぶ病抵抗反応におけるカブ培養根の細胞内Ca2+ 濃度（［Ca2+］cyt）の変動を調査するため、Ca2+ の蛍光指示薬Fura-2のアセトキシメチルエステル（Fura-2/AM）をカブ培養根に導入し、蛍光顕微鏡により［Ca2+］cyt を定量的に解析することを試みた。蛍光顕微鏡下で細胞質内にFura-2が導入されたことが確認できたカブ培養根に、カルシウムイオノフォアA23187とCa2+ を同時に処理すると、直ちに［Ca2+］cyt の相対値を示すFura-2蛍光比が上昇したことから、Fura-2/AM を用いた本手法は、培養根における［Ca2+］cyt の変化を観察するのに適していることが示された。さらに［Ca2+］cyt を増加させることが知られている外的刺激として、マンニトール及びNaCl の高濃度処理を行ったところ、同様なFura-2蛍光比の急激な増加が観察され、本手法の汎用性が示された。次に、我々は本来の目的である根こぶ病抵抗反応における［Ca2+］cyt の解析に本手法を応用した。Fura-2/AM を導入した根こぶ病抵抗性カブ培養根にP. brassicae の休眠胞子を処理しても、500秒の測定時間内に［Ca2+］cyt が変化することは無かった。しかし休眠胞子を発芽促進液（GES: germination enhancing suspension）で前処理し、同様の実験を行ったところ、非常にわずかであったものの、再現性ある［Ca2+］cyt の上昇が観察された。根こぶ病に対する初期抵抗反応におけるCa2+ の重要性を立証するためには、今後さらなる解析が必要であるが、その過程において本手法は大きく貢献するものと期待される。","subitem_description_type":"Abstract"}]},"item_7_full_name_3":{"attribute_name":"著者別名","attribute_value_mlt":[{"nameIdentifiers":[{"nameIdentifier":"163525","nameIdentifierScheme":"WEKO"}],"names":[{"name":"高橋, 秀行"}]},{"nameIdentifiers":[{"nameIdentifier":"163526","nameIdentifierScheme":"WEKO"}],"names":[{"name":"石川, 寿樹"}]},{"nameIdentifiers":[{"nameIdentifier":"163527","nameIdentifierScheme":"WEKO"}],"names":[{"name":"早川, 徹"}]},{"nameIdentifiers":[{"nameIdentifier":"163528","nameIdentifierScheme":"WEKO"}],"names":[{"name":"伊藤, 紀美子"}]},{"nameIdentifiers":[{"nameIdentifier":"163529","nameIdentifierScheme":"WEKO"}],"names":[{"name":"三ツ井, 敏明"}]},{"nameIdentifiers":[{"nameIdentifier":"163530","nameIdentifierScheme":"WEKO"}],"names":[{"name":"堀, 秀隆"}]}]},"item_7_publisher_7":{"attribute_name":"出版者","attribute_value_mlt":[{"subitem_publisher":"新潟大学農学部"}]},"item_7_select_19":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_select_item":"publisher"}]},"item_7_source_id_11":{"attribute_name":"書誌レコードID","attribute_value_mlt":[{"subitem_source_identifier":"AN00183393","subitem_source_identifier_type":"NCID"}]},"item_7_source_id_9":{"attribute_name":"ISSN","attribute_value_mlt":[{"subitem_source_identifier":"03858634","subitem_source_identifier_type":"ISSN"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"Takahashi, Hideyuki"}],"nameIdentifiers":[{"nameIdentifier":"163519","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"Ishikawa, Toshiki"}],"nameIdentifiers":[{"nameIdentifier":"163520","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"Hayakawa, Tohru"}],"nameIdentifiers":[{"nameIdentifier":"163521","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"Itoh, Kimiko"}],"nameIdentifiers":[{"nameIdentifier":"163522","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"Mitsui, Toshiaki"}],"nameIdentifiers":[{"nameIdentifier":"163523","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"Hori, Hidetaka"}],"nameIdentifiers":[{"nameIdentifier":"163524","nameIdentifierScheme":"WEKO"}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2019-08-20"}],"displaytype":"detail","filename":"1_0009.pdf","filesize":[{"value":"208.1 kB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"1_0009.pdf","url":"https://niigata-u.repo.nii.ac.jp/record/28421/files/1_0009.pdf"},"version_id":"8d580c25-59e6-4b34-a9cc-8a37e6daeadc"}]},"item_keyword":{"attribute_name":"キーワード","attribute_value_mlt":[{"subitem_subject":"Brassica rapa","subitem_subject_scheme":"Other"},{"subitem_subject":"clubroot resistance","subitem_subject_scheme":"Other"},{"subitem_subject":"cytosolic [Ca2+]","subitem_subject_scheme":"Other"},{"subitem_subject":"Fura-2","subitem_subject_scheme":"Other"},{"subitem_subject":"root culture","subitem_subject_scheme":"Other"},{"subitem_subject":"Brassica rapa","subitem_subject_scheme":"Other"},{"subitem_subject":"Fura-2","subitem_subject_scheme":"Other"},{"subitem_subject":"細胞内Ca2+ 濃度","subitem_subject_scheme":"Other"},{"subitem_subject":"根こぶ病抵抗性","subitem_subject_scheme":"Other"},{"subitem_subject":"培養根","subitem_subject_scheme":"Other"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"eng"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"departmental bulletin paper","resourceuri":"http://purl.org/coar/resource_type/c_6501"}]},"item_title":"Loading of Fura-2 into Liquid Organ Cultured Adventitious Root of Turnip (Brassica rapa L.) Resistant to Clubroot Pathogen Plasmodiophora brassicae and Determination of [Ca2+]cyt","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"Loading of Fura-2 into Liquid Organ Cultured Adventitious Root of Turnip (Brassica rapa L.) Resistant to Clubroot Pathogen Plasmodiophora brassicae and Determination of [Ca2+]cyt"},{"subitem_title":"Loading of Fura-2 into Liquid Organ Cultured Adventitious Root of Turnip (Brassica rapa L.) Resistant to Clubroot Pathogen Plasmodiophora brassicae and Determination of [Ca2+]cyt","subitem_title_language":"en"}]},"item_type_id":"7","owner":"1","path":["456","1602"],"pubdate":{"attribute_name":"公開日","attribute_value":"2007-10-12"},"publish_date":"2007-10-12","publish_status":"0","recid":"28421","relation_version_is_last":true,"title":["Loading of Fura-2 into Liquid Organ Cultured Adventitious Root of Turnip (Brassica rapa L.) Resistant to Clubroot Pathogen Plasmodiophora brassicae and Determination of [Ca2+]cyt"],"weko_creator_id":"1","weko_shared_id":null},"updated":"2022-12-15T03:58:27.855242+00:00"}