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Elucidation of the new mechanism for preventing reactive oxygen species production depending on chloroplastic ATP synthase in land plants

阐明陆地植物中依靠叶绿体 ATP 合酶防止活性氧产生的新机制

基本信息

批准号：
18J00852
负责人：
高木大輔
金额：
$ 2.58万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for JSPS Fellows
财政年份：
2018
资助国家：
日本
起止时间：
2018-04-25 至 2021-03-31
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18J00852/
关键词：
リン酸 ATPアーゼ光合成植物栄養学イネ植物生理学 Rubisco 光傷害

项目摘要

野生型イネ(ノトヒカリ)を異なる無機リン酸(Pi)施肥条件下で栽培し、無機リン酸量を増大させた生葉における光合成駆動時の葉緑体局在型ATPシンターゼの回転活性(gH+)は、高リン酸施肥により葉身Pi量が増加する条件で低下することを明らかとした。本年度は同条件下において光合成駆動時の代謝物の定量を行うことでATPシンターゼの回転活性低下の原因解明を試みた。ノトヒカリをリン酸施肥条件を5区画(低Pi条件,標準Pi条件、標準2倍量Pi施肥条件、標準3倍量Pi施肥条件、標準4倍量Pi施肥条件、標準5倍量Pi施肥条件)設定し、水耕栽培条件下で生育させた。発芽後70日以降の最上位展開葉を明条件下でサンプリングした後に、早急に液体窒素にて凍結させ光合成駆動時の代謝反応を即時に停止させた。サンプリングした葉は、代謝物抽出後にトリプル四重極キャピラリー電気泳動質量分析装置により各種代謝物の定量を行った。これに加え、ATPシンターゼ活性変異体の作成は、ノトヒカリを用い、gH+活性制御に関わると考えられるγ-subunitのATPシンターゼ過剰発現体及びゲノム編集体の作成を行った。代謝物解析の結果、ATP、ADP、AMPの全量はリン酸施肥量の増加に伴って増加することが明らかとなったが、光合成駆動時のATP/ADP比の算出を行うとリン酸施肥量の増加に伴ってATP/ADP比が有意に増大することが明らかとなった。このATPが余剰する状況は、前年度に明らかとしたRubiscoの活性化率低下に伴うカルビンサイクルの回転速度の低下と一致するデータである。これらの結果から、gH+はこれまで報告されていたような単純に細胞内のPi量によって変動するだけではなく、葉緑体内部の代謝回転をATP/ADP比の変化で検知することで変化していることが新規に明らかとなった。

Under the condition of fertilization with no organic acid (Pi), under the condition of fertilization with no organic acid (Pi), the photosynthesis activity (gH+) of the plant under the condition of fertilization of organic acid (Pi) was higher than that of the plant under the condition of fertilization of organic acid (Pi). Under the condition of fertilization, the photosynthesis activity (gH+) of the plant under the condition of high concentration of organic acid and high concentration of organic acid was significantly increased. Under the same conditions this year, under the same conditions, the reason for the low activity of photosynthesis was explained by quantitative analysis of the metabolites of photosynthesis during the period of photosynthesis. The reason for the low activity of ATP was analyzed in this year. The conditions of acid fertilization were set in 5 areas (low Pi condition, standard Pi condition, standard 2-fold Pi fertilization condition, standard 3-fold Pi fertilization condition, standard 4-fold Pi fertilization condition, standard 5-fold Pi fertilization condition) and water-tillage cultivation. 70 days after sprouting, the uppermost part of the plant was opened in the first place under the condition of light, and the photosynthesis of photosynthesis was stopped immediately under the condition of early and urgent liquid asphyxiation. After the substitutes and substitutes have been extracted, the equipment for the quantitative analysis of swimming movements has been checked by the equipment for quantitative analysis of each substitute. In addition, the ATP system is used to make the system, the system is used, and the gH+ activity system is used. The γ-subunit ATP system is used to improve the performance of the system. The results of substitute analysis, ATP, ADP, AMP were used to analyze the results, and the results of ATP, ATP/ADP and ATP/ADP were calculated by the results of substitute analysis, the results of the results of substitute analysis, the results of total acid fertilizer application, ATP/ADP, total acid fertilizer application, total acid fertilizer, total acid fertilizer and ATP/ADP. The previous year showed that the activity rate of ATP was low, and that the activity rate of Rubisco was low in the previous year. The results of the experiment, the results of the gH+ report, the volume of the cell, the number of cycles in the body, the response rate of the ATP/ADP in the body, and the information on the new rules and regulations of the system.