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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 洪傳揚(Chwan-Yang Hong) | |
dc.contributor.author | Wan-Rung Lin | en |
dc.contributor.author | 林婉容 | zh_TW |
dc.date.accessioned | 2021-06-08T04:18:17Z | - |
dc.date.copyright | 2010-07-28 | |
dc.date.issued | 2010 | |
dc.date.submitted | 2010-07-27 | |
dc.identifier.citation | Abogadallah GM, Serag MM, Quick WP (2010) Fine and coarse regulation of reactive oxygen species in the salt tolerant mutants of barnyard grass and their wild-type parents under salt stress. Physiologia Plantarum 138: 60-73
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/22459 | - |
dc.description.abstract | 鹽分逆境常誘導活化氧族(reactive oxygen species , ROS)產生,對植物造成氧化傷害,植物為了降低鹽分逆境下的氧化傷害發展出許多應對的策略,如ascorbate-glutathione cycle(ASC-GSH cycle),其中 glutathione reductase(GR)在 ASH-GSH cycle 中扮演重要角色,負責將氧化態的GSSG 還原為GSH。水稻GR 基因家族由三個基因組成,分別為細胞質的OsGR2,和葉綠體的 OsGR1及OsGR3。我們過去的研究發現 OsGR3 在水稻幼苗根部受 NaCl 和 abscisic acid 誘導表現,但是其胺基酸序列由於缺失了重要的 FAD 結合功能區塊,被認為是一個不具有功能的 GR 同功酵素。本試驗透過水稻序列資料庫比對 OsGR3 序列後,發現在預測序列 N 端應存在一個 FAD 結合功能區塊,為進一步了解此一全長的 OsGR3 是否具有GR 活性,及鹽分逆境對 OsGR3 的反應,本試驗透過大量表現 OsGR3 及osgr3 突變體研究其功能。結果顯示, osgr3 突變株會降低 42% 的 GR 活性,同時幼苗表現出對鹽分敏感的外表型,處理鹽分一週後 osgr3 存活率僅為 9%,在第二片葉片會累積較多的 H2O2,並降低 Fv/Fm。進一步分析發現,osgr3 幼苗在鹽分逆境下,ascorbate (ASC) 含量減少,且ASC/DHA(dehydroascorbate)降低,推測這可能是導致 osgr3 不耐鹽分逆境之原因。在大量表現 OsGR3 植株中,GR 活性提高 3∼6 倍,並比 WT 有較高鹽分耐受性,且大量表現 OsGR3 之轉殖株含有較高的 ASC 含量。而在大量表現 FAD 缺失的OsGR3tc 轉殖株中,GR 活性降低,且植株不耐鹽。另一方面,將 OsGR3 轉殖回 osgr3 的功能補償轉殖株則可增加 3∼5 倍 GR 活性降低,並回復對高鹽逆境的耐受性。此外,以 methyl viologen (MV) 處理水稻葉圓片,結果顯示抵抗葉綠素降解能力以 osgr3 功能補償植株最佳,其次為 WT,而以 osgr3 居於最後,顯示OsGR3和水稻抗氧化逆境有關。綜合以上結果,OsGR3不但具有 GR 活性,並與水稻的鹽分耐受性及抗氧化逆境有關。 | zh_TW |
dc.description.abstract | Salt stress results in an excessive generation of reactive oxygen species (ROS) that causes oxidative damage to plants. Plants evolve certain strategies to remove salt-induced reactive oxygen species (ROS), such as glutathione-ascorbate cycle (ASH-GSH cycle). Glutathione reductase (GR) plays an important role in ASC-GSH cycle, which converted oxidized GSH (GSSG) to reduced GSH. Three GR genes exist in rice, including a cytosolic (OsGR2) and two chloroplastic isoforms (OsGR1 and OsGR3). Our previous study demonstrated that expression of OsGR3 was increased in response to NaCl and abscisic acid in roots of etiolated rice seedlings. The OsGR3 has been considered to be a non-functional GR isozyme due to the lack of a FAD binding domain and chloroplastic transit sequence (CTS). However, after blasting different rice database, we found a full length OsGR3 open reading frame with complete FAD binding domain and CTS. To investigate the physiological roles of OsGR3 in salt stress and to understand whether the OsGR3 is a functional GR, OsGR3 knockout mutant and overexpression transgenic rice were analyzed in this thesis. In osgr3, 42% GR activity was decreased and salt sensitivity was enhanced. Physiological responses of osgr3 rice mutant revealed that the maximal efficiency of photosystem II and survival rate were notably reduced as compared to WT, and H2O2 was increased after 200 mM NaCl treatment. One of the reasons caused osgr3 mutant intolerant to salt stress is that osgr3 had lower ascorbate content and lower ratio of ascobate/dehydroascorbate than WT rice. Overexpression of OsGR3 in rice plants increased three to six times of GR activity, and also increased transformant's salt tolerance. Higher ascorbate content in overexpression OsGR3 transgenic rice as compared to WT was also observed. However, overexpression of OsGR3tc, lack of FAD binding domain, diminished GR activity, and transgenic rice were sensitive to salt stress. On the other hand, complementation of osgr3 with OsGR3 (osgr3/OsGR3) increased three to five times of GR activity and restored salt stress tolerance. Meanwhile, leaf discs of osgr3, WT and complementation treated with methyl viologen showed that the degrees of chlorophyll bleaching from osgr3 to WT then osgr3/OsGR3, indicating that OsGR3 is important for oxidative defense system. Our results suggested that OsGR3 is a functional GR, and is involved in salt and oxidative tolerance. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T04:18:17Z (GMT). No. of bitstreams: 1 ntu-99-R97623003-1.pdf: 6810771 bytes, checksum: 6c8d9ed28ff6e2d0e95b5b403d23f381 (MD5) Previous issue date: 2010 | en |
dc.description.tableofcontents | 誌謝 ii
中文摘要 iv ABSTRACT v 縮寫字對照 vii 目錄 viii 圖目錄 xi 表目錄 xiii 附表目錄 xiii 壹、 前人研究 1 一、 非生物逆境對作物產量及品質的影響 1 二、 氧化逆境對植物生長發育的影響 1 三、 植物的抗氧化機制 2 四、 鹽分逆境對植物生長發育的影響 4 五、 植物Glutathione reductase(GR)的生化特性與生理功能 5 5.1 GR 的生化特性 6 5.2 GR 的構造 6 5.3 GR 基因及其相似度(homology) 7 5.4 GR 的同功酵素(isoenzyme) 8 5.5 GR活性與植物逆境耐受性的關係 8 5.6 GR 在植物的功能 10 六、 水稻 OsGR 基因家族特性分析 13 貳、 本論文研究目的及實驗架構 15 參、 材料與方法 16 一、 植物材料的準備及生長條件 16 二、 質粒的構築 16 三、 水稻基因轉殖 21 四、 基因表現分析材料準備及處理 23 五、 基因表現分析 27 六、 基因型分析(genotyping) 28 七、 水稻逆境處理及生理分析 29 八、 GR同功酵素染色分析 34 九、 啟動子特性分析 34 十、 統計分析 35 肆、 結果 36 一、 水稻 GR3 分子特性分析 36 1.1 OsGR3 基因基因結構分析 36 1.2 OsGR3 基因組織專一性表現 36 1.3 OsGR3 啟動子的特性分析 37 二、 OsGR3基因剔除突變株的功能分析 38 2.1 OsGR3 基因Tos17 突變株的分子鑑定 38 2.2 失去OsGR3降低水稻植株對鹽分及氧化逆境的耐受性 39 2.3 以OsGR3 補償突變株 osgr3 之水稻植株可以恢復植株對鹽分逆境及氧化逆境的耐受性。 41 2.4 大量表現OsGR3及 OsGR3tc轉殖株分析 42 2.5 大量表現OsGR3 及OsGR3tc 轉殖株對鹽分逆境及氧化逆境的耐受性 43 伍、 討論 44 一、 鹽分逆境會增加 GR 活性 44 二、 水稻 GR3 分子特性分析 44 三、 OsGR3 的組織專一性及啟動子的特性分析 44 四、 水稻 GR3 是一個有活性的 Glutathione Reductase,而 OsGR3tc 則否 46 五、 OsGR3 參與水稻對鹽分逆境的耐受性的調控 48 六、 OsGR3 參與水稻對氧化逆境耐受性的調控 51 陸、 參考文獻 52 柒、 附錄 85 一、 水稻基因轉殖用培養基列表 85 二、 木村氏(Kimura)水耕液配方(helf-strength) 88 三、 原態膠體電泳分析 89 四、 試驗所用載體(利用 vector NTI 繪製) 91 | |
dc.language.iso | zh-TW | |
dc.title | Glutathione Reductase 3 對水稻鹽分逆境功能之研究 | zh_TW |
dc.title | Studies on the role of glutathione reductase 3 in salt stress of rice | en |
dc.type | Thesis | |
dc.date.schoolyear | 98-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 高景輝(Ching-Huei Kao),李澤民(Tse-Min Lee),葉國楨(Kuo-Chen Yeh),張孟基(Men-Chi Chang) | |
dc.subject.keyword | 穀胱甘肽,過還原酶,抗氧化酵素,鹽分逆境,氧化逆境,轉殖水稻, | zh_TW |
dc.subject.keyword | glutathione reductase,antioxidative enzymes,salt stress,oxidative stress,transgenic plant, | en |
dc.relation.page | 94 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2010-07-28 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 農業化學研究所 | zh_TW |
顯示於系所單位: | 農業化學系 |
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