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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 葉開溫 | |
| dc.contributor.author | Hsin-Yi Lin | en |
| dc.contributor.author | 林欣儀 | zh_TW |
| dc.date.accessioned | 2021-06-16T02:29:06Z | - |
| dc.date.available | 2018-08-03 | |
| dc.date.copyright | 2015-08-03 | |
| dc.date.issued | 2015 | |
| dc.date.submitted | 2015-07-31 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/53760 | - |
| dc.description.abstract | 前人研究指出高溫誘導文心蘭開花與維生素C (ascorbate, AsA)及穀胱甘肽(glutathione, GSH) 的氧化還原狀態相關，並受AsA-GSH循環及穀胱甘肽生合成路徑影響。其參與酵素包含穀胱甘肽還原酶glutathione reductase (OgGR), γ-ECS,γ-glutamylcysteine synthetase (OgGSH1)及穀胱甘肽合成酶glutathione synthase (OgGSH2)。文心蘭GIGANTEA (GI) 及 FLOWERING LOCUS T (FT) 等開花基因的表現量亦顯著上升。然而，高溫誘導文心蘭開花路徑中，穀胱甘肽氧化還原狀態如何調控開花基因的表現，目前尚不明瞭。首先，我們利用氧化態維生素C處理文心蘭，結果可使穀胱甘肽還原態下降、氧化態上升、脫氫維他命C還原酶活性上升，所以可確定AsA-GSH循環存在於文心蘭中。接著，我們利用文心蘭穀胱甘肽還原酶(OgGR), γ-ECS (OgGSH1)及穀胱甘肽合成酶(OgGSH2)等基因，分別過表現於阿拉伯芥，結果顯示經30度高溫處理後，過表現轉殖株較WT延遲開花，且穀胱甘肽還原酶活性下降、穀胱甘肽氧化還原比例會隨時間提高，而WT則下降。反之在阿拉伯芥之穀胱甘肽代謝相關突變株，cad2、pad2 及gr1，與過表現轉殖株作比較之結果顯示，經30度高溫處理後，pad2, cad2, gr1有提早開花的情形，且穀胱甘肽還原酶活性及穀胱甘肽氧化還原比例均顯著下降。因此可知，雖然穀胱甘肽氧化還原比例於過表現轉植株及突變株本身即不相同，但處理高溫後即可明顯看到開花與穀胱甘肽氧化還原比例存有正向關聯性。此外，我們亦釣取文心蘭GI的啟動子，並預測感興趣的cis-element，結果說明文心蘭的GI可能被環境逆境調控，故推測文心蘭的GI和環境逆境所產生的氧化還原變化有交互作用。文心蘭的GI蛋白表現於細胞質及細胞核，說明文心蘭的GI具有調控下游開花基因的潛力。根據實驗結果推測，高溫造成的穀胱甘肽氧化還原變化並使文心蘭提早開花的路徑，是藉由穀胱甘肽的生合成及代謝途徑，進而影響下游GI的表現所造成的結果。 | zh_TW |
| dc.description.abstract | Previous study reveals that high ambient temperature make Oncidesa early flowering through changing ascorbate (AsA) and glutathione (GSH) redox status (AsA/DHA) (GSH/GSSG) via AsA-GSH cycle and GSH biosynthesis. Glutathione reductase (OgGR), γ-glutamylcysteine synthetase (OgGSH1) and glutathione synthase (OgGSH2), as well as floral genes, such as GIGANTEA (GI) and FLOWERING LOCUS T (FT) are related to this pathway. However, the mechanism of GSH redox regulated floral genes on high temperature-induced flowering in Oncidesa is still unknown. First, we treat Oncidesa with dehydroascorbate for 14 days and find glutathione content decrease and glutathione disulfide content increase and dehydroascorbate reductase activity increase. Therefore, we can confirm that the AsA-GSH cycle exist in Oncidesa. Thus, we over-express OgGR, OgGSH1, and OgGSH2 in Arabidopsis to study the role of GSH on high temperature-induced flowering. The results show the overexpression plant delay flowering and increase of GSH redox. Otherwise, GSH metabolism related mutants in Arabidopsis, cad2, pad2, and gr1 which are opposite to the overexpression lines, showed early flowering with low GSH content, indicating GSH redox status and which key regulating enzyme involved in high-temperature induced flowering. We clone GI promoter from Oncidesa, and predict the cis-element, revealing that GI is probably regulated by stress, and implies the interaction between GI and redox signaling. Moreover, Oncidesa GI is located in nucleus, indicating that GI potentially regulate downstream floral gene. We suggest that the change of GSH redox ratio under high-temperature can induce early flowering via GSH biosynthesis/metabolism and influence the gene expression of GI in Oncidesa. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T02:29:06Z (GMT). No. of bitstreams: 1 ntu-104-R02b42010-1.pdf: 3823142 bytes, checksum: 3a8ad710b0384ea7ca1dbabcbad48f90 (MD5) Previous issue date: 2015 | en |
| dc.description.tableofcontents | 目錄
中文摘要.....................................................................................................................1 英文摘要.......................................................................................................................2 目錄...............................................................................................................................3 圖目錄...........................................................................................................................5 附錄目錄......................................................................................................................7 第一章前言.................................................................................................................8 第一節文心蘭概述.....................................................................................................8 第二節植物於氧化逆境下的反應與抗氧化物質...................................................10 第三節植物的開花路徑與開花基因.......................................................................17 第四節前人研究回顧及研究目的...........................................................................20 第二章、材料與方法.................................................................................................22 第一節實驗材料.......................................................................................................22 第二節實驗方法.......................................................................................................22 第三章結果............................................................................................................... 32 第一節文心蘭中維生素C相關藥劑處理對穀胱甘肽氧化還原之變化................33 第二節文心蘭OgGSH1, OgGSH2, OgGR全長序列的分析...................................33 第三節文心蘭OgGSH1, OgGSH2, OgGR之表現位置...........................................34 第四節文心蘭OgGSH1, OgGSH2, OgGR過表現之阿拉伯芥轉殖株建構及分析 ....................................................................................................................36 第五節溫度處理對文心蘭OgGSH1, OgGSH2, OgGR之過表現阿拉伯芥轉殖株之 穀胱甘肽氧化還原狀態變化與開花時間的關係....................................38 第六節 溫度處理對阿拉伯芥穀胱甘肽突變株之穀胱甘肽氧化還原狀態變化與開 花時間的關係............................................................................................41 第七節文心蘭GIGANTEA可能參與之開花調控...................................................43 第四章討論............................................................................................................... 45 第一節 文心蘭維生素C氧化還原狀態與穀胱甘肽C氧化還原變化之關係.........45 第二節文心蘭OgGSH1, OgGSH2, OgGR 蛋白功能探討與穀胱甘肽含量及氧化 還原狀態變化之連結對開花的關係........................................................46 第三節溫度對穀胱甘肽氧化還原狀態變化及開花的影響...................................48 第四節GSH1, GSH2, GR影響穀胱甘肽氧化還原狀態對開花的影響..................50 第五節溫度及GSH1, GSH2, GR間的交感作用調控開花......................................52 第六節文心蘭GIGANTEA可能參與之開花調控...................................................54 第七節未來展望.......................................................................................................55 參考文獻..................................................................................................................... 56 圖目錄 圖一、還原態(AsA) 及氧化態(DHA) 維生素C處理對文心蘭假球莖中穀胱甘 肽含量及酵素活性變化的情形......................................................................71 圖二、文心蘭GSH1之保守區域分析、胺基酸編碼序列比對及親緣演化樹分析...72 圖三、文心蘭GSH2之保守區域分析、胺基酸編碼序列比對及親緣演化樹分析...73 圖四、文心蘭GR之保守區域分析、胺基酸編碼序列比對及親緣演化樹分析... 74 圖五、文心蘭GSH1, GSH2, GR蛋白之次細胞表現位置......................................75 圖六、文心蘭GSH1, GSH2, GR基因之過表現載體建構........................ ................76 圖七、文心蘭GSH1, GSH2, GR基因過表現於阿拉伯芥植株之基因表現量分析77 圖八、檢測文心蘭GSH1, GSH2, GR基因過表現於阿拉伯芥植株之穀胱甘肽含量 及酵素活性.................................................................................................... .78 圖九、文心蘭GSH1, GSH2, GR基因過表現於阿拉伯芥植株之表型觀察......... ...79 圖十、文心蘭GSH1, GSH2, GR基因過表現於阿拉伯芥突變株cad2, pad2, gr1之穀 胱甘肽含量及酵素活性................................................................................. 80 圖十一、22度短日照下文心蘭GSH1, GSH2, GR基因過表現於阿拉伯芥植株之開 花時間的影響............................................................................................. 81 圖十二、高溫處理對文心蘭GSH1, GSH2, GR基因過表現於阿拉伯芥植株之開花 時間的影響................................................................................................. 82 圖十三、溫度處理後文心蘭GSH1, GSH2, GR基因過表現於阿拉伯芥植株之酵素 活性............................................................................................................. 83 圖十四、溫度處理後文心蘭GSH1, GSH2, GR基因過表現於阿拉伯芥植株之穀胱 甘肽氧化及還原態含量............................................................................. 84 圖十五、溫度處理後文心蘭GSH1, GSH2, GR基因過表現於阿拉伯芥植株之穀胱 甘肽氧化還原比例變化............................................................................. 85 圖十六、溫度處理對阿拉伯芥突變株之開花時間的影響..................................... 86 圖十七、溫度處理後阿拉伯芥突變株之酵素活性................................................. 87 圖十八、溫度處理後阿拉伯芥突變株之穀胱甘肽含量......................................... 88 圖十九、文心蘭GI蛋白之次細胞表現位置............................................................ 89 圖二十、文心蘭GI啟動子釣取................................................................................ 90 附錄目錄 附錄一、文心蘭Gower Ramsey品系親緣圖譜........................................................ 91 附錄二、文心蘭Gower Ramsey之生長週期............................................................ 92 附錄三、維生素C-穀胱甘肽循環(AsA-GSH cycle).............................................. 93 附錄四、穀胱甘肽 (GSH) 生合成路徑示意圖...................................................... 94 附表一、植物重要的抗氧化酵素............................................................................. 95 附表二、引子序列..................................................................................................... 96 附表三、檢索表......................................................................................................... 97 | |
| dc.language.iso | zh-TW | |
| dc.subject | 文心蘭 | zh_TW |
| dc.subject | 維他命Ｃ-穀胱甘?循環 | zh_TW |
| dc.subject | 穀胱甘?氧化還原比例 | zh_TW |
| dc.subject | 開花 | zh_TW |
| dc.subject | 高溫 | zh_TW |
| dc.subject | Oncidesa | en |
| dc.subject | AsA-GSH cycle | en |
| dc.subject | GSH redox | en |
| dc.subject | flowering | en |
| dc.subject | high temperature | en |
| dc.title | 穀胱甘肽氧化還原狀態影響文心蘭的高室溫誘導
開花機制 | zh_TW |
| dc.title | Glutathione redox status affects the high ambient
temperature-induced flowering in Oncidesa | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 103-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 陳虹樺,詹明才,王淑珍 | |
| dc.subject.keyword | 維他命Ｃ-穀胱甘?循環,穀胱甘?氧化還原比例,開花,高溫,文心蘭, | zh_TW |
| dc.subject.keyword | AsA-GSH cycle,GSH redox,flowering,high temperature,Oncidesa, | en |
| dc.relation.page | 97 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2015-08-03 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 植物科學研究所 | zh_TW |
| 顯示於系所單位： | 植物科學研究所 | |
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