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dc.contributor.advisor洪傳揚(Chwan-Yang Hong)
dc.contributor.authorSzu-Yun Chenen
dc.contributor.author陳思昀zh_TW
dc.date.accessioned2021-06-16T08:47:14Z-
dc.date.available2018-08-28
dc.date.copyright2013-08-28
dc.date.issued2013
dc.date.submitted2013-08-20
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/59056-
dc.description.abstract硫氧化還原蛋白(Thioredoxin, 簡稱Trx)是一類利用硫基(Thiol group)氧化還原反應(2SH←→S-S)調節細胞氧化還原狀態的蛋白,與其專一性的Trx還原酶(The NADPH-dependent thioredoxin reductase, 簡稱NTR)構成NTR/Trx系統,在植物氧化逆境抗性反應中扮演重要的角色。Sulfolobus solfataricus是一種生長在極端環境的古生菌,為了解此系統對植物逆境抗性之影響,本研究將S. solfataricus 兩個Trx基因SsTrxA1、SsTrxA2,以及一個NTR基因SsTrxB3,分別利用35S啟動子驅動構築成可在植物大量表現的質粒,包括OE-SsTrxA1、OE-SsTrxA2、OE-SsTrxB3 、OE- SsTrxA1+ SsTrxB3、OE- SsTrxA2+SsTrxB3等5個質粒,並利用農桿菌轉殖大量表現在阿拉伯芥,以分析轉殖植物對逆境之抗性。RT-PCR分析結果顯示,五種質粒均可成功的大量表現在阿拉伯芥植株中,各基因構築均可獲得6個以上轉殖系。轉殖株經過鎘、巴拉刈處理後,顯示SsTrxB3 與 SsTrxA2+SsTrxB3轉殖株有較佳的逆境抗性,在逆境下與未轉殖植株比較,有較長的根長與較大的鮮重。此外,熱逆境分析結果顯示OE- SsTrxA2B3具有較佳的先天耐熱性。這些結果顯示來自S. solfataricus的NTR/Trx系統能提高植物的逆境抗性,未來應具有潛力應用於增加作物耐多重逆境抗性。zh_TW
dc.description.abstractNADPH-dependent thioredoxin reductase/Thioredoxin (NTR/Trx) system catalyzes disulfide-bond reduction to regulate the redox status of cells, ubiquitously exists in all organisms and involves in oxidative stress response. The NTR/Trx system of a hyperthermophilic archaeon, Sulfolobus solfataricus, has been found to play a major role in its extremely thermal tolerance. To study the impact of S. solfataricus NTR/Trx system on plant abiotic stress tolerance, S. solfataricus NTR/Trx system genes (two Trx genes:SsTrxA1, SsTrxA2 and one NTR gene :SsTrxB3) were solely or simultaneously overexpressed in Arabidopsis thaliana to evaluate their roles in abiotic stress tolerance. Five plasmids including SsTrxA1, SsTrxA2, SsTrxB3, SsTrxA1+ SsTrxB3 and SsTrxA2+SsTrxB3 under the control of a CaMV 35S promoter were constructed and transformed to Arabidopsis by Agrobacterium tumefaciens. At least 6 transgenic lines were obtained for each construct and most of them showed highly accumulation of NTR or Trx mRNA. Functional analysis indicated that OE-SsTrxB3 and OE-SsTrxA2B3 plants exhibited enhanced tolerance of transgenic plants to Cd and paraquat.Moreover, SsTrxA2+SsTrxB3 plants also showed increased thermal tolerance. Taken together, the S. solfataricus NTR/Trx system is able to enhance stress tolerance in Arabidopsis. This system may be applied for the development of new strategies to enhance crop tolerance to multiple stresses.en
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Previous issue date: 2013
en
dc.description.tableofcontents第一章、 緒論 1
1.1 植物與氧化逆境 1
1.1.1 氧化逆境之來源與影響 1
1.1.2 鎘(Cadmium, Cd)對植物之氧化毒性 2
1.1.3 巴拉刈(paraquat, PQ)對植物之氧化毒性 3
1.1.4 高溫對植物之氧化毒性 3
1.1.5 植物的抗氧化系統:酵素型、非酵素 4
1.1.6 植物的細胞氧化還原狀態調節系統:NADP+硫氧化還原蛋白還原酶/硫氧化還原蛋白系統(NTR/Trx系統)、榖胱甘肽/穀氧化還原蛋白(GSH/Grx系統) 4
1.2 阿拉伯芥的NTR/Trx系統 8
1.2.1 阿拉伯芥介紹 8
1.2.2 Trx 9
1.2.3 NTR 10
1.2.4 植物NTR/Trx系統對細胞氧化還原狀態調控的影響 11
1.3 Sulfolobus solfataricus NTR/Trx系統 11
1.3.1 Sulfolobus solfataricus介紹 11
1.3.2 Trx 12
1.3.3 NTR 12
第二章、 研究目的 14
第三章、 材料與方法 15
3.1 試驗材料 15
3.2 胺基酸序列比對及親緣演化樹分析 15
3.3 質粒的構築 15
3.4 載體或DNA片段的製備 17
3.5 大腸桿菌(E.coli)質粒DNA的純化 19
3.6 阿拉伯芥之轉殖 20
3.7 轉殖基因表現分析 22
3.8 生理分析 26
第四章、 結果 28
4.1 硫葉菌(S. solfataricus)TrxA1、TrxA2與TrxB3之結構及親緣演化樹分析 28
4.1.1 SsTrxA1、SsTrxA2與阿拉伯芥典型Trx胺基酸序列比對及親緣演化樹分析 28
4.1.2 SsTrxB3與其他物種的NTR胺基酸序列比對及親緣演化樹分析 29
4.2 大量表現SsTrxA1、SsTrxA2、SsTrxB3的A. thaliana轉殖株之篩選及分子鑑定 29
4.2.1 SsTrxA1、SsTrxA2、SsTrxB3基因於A. thaliana轉殖株之表現質粒之構築 29
4.2.2 SsTrxA1、SsTrxA2、SsTrxB3基因於A. thaliana轉殖株之分子表現鑑定 30
4.3 轉殖株幼苗之鎘逆境抗性檢測 30
4.3.1 植株外表觀察 30
4.3.2 根長、鮮重分析 30
4.3.3 葉綠素含量分析 31
4.3.4 H2O2含量分析 31
4.4 轉殖株幼苗巴拉刈(PQ)逆境抗性檢測 31
4.4.1 植株外表觀察 31
4.5 轉殖株幼苗熱逆境抗性檢測 31
4.5.1 轉殖株耐熱性性狀分析 32
4.5.2 轉殖株耐熱性存活率分析 32
第五章、 討論 33
第六章、 參考文獻 38
附錄 65
dc.language.isozh-TW
dc.title在阿拉伯芥大量提升耐熱古生菌Sulfolobus solfataricus 硫氧化還原系統對非生物性逆境抗性之研究zh_TW
dc.titleStudies on the increase of abiotic stress tolerance in Arabidopsis thaliana by enhancing thioredoxin system from a hyperthermophilic archaeon Sulfolobus solfataricusen
dc.typeThesis
dc.date.schoolyear101-2
dc.description.degree碩士
dc.contributor.oralexamcommittee張孟基(Men-Chi Chang),葉信宏(Hsin-Hung Yeh),陳仁治(Jen-Chih Chen),徐駿森(Chun-Hua Hsu)
dc.subject.keyword硫葉菌,阿拉伯芥,鎘,氧化逆境,硫氧化還原蛋白,zh_TW
dc.subject.keywordS. solfataricus,A. thaliana,Cd,oxidative stress,thioredoxin,thioredoxin reductase,en
dc.relation.page67
dc.rights.note有償授權
dc.date.accepted2013-08-20
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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