請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/59056
標題: | 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 Studies on the increase of abiotic stress tolerance in Arabidopsis thaliana by enhancing thioredoxin system from a hyperthermophilic archaeon Sulfolobus solfataricus |
作者: | Szu-Yun Chen 陳思昀 |
指導教授: | 洪傳揚(Chwan-Yang Hong) |
關鍵字: | 硫葉菌,阿拉伯芥,鎘,氧化逆境,硫氧化還原蛋白, S. solfataricus,A. thaliana,Cd,oxidative stress,thioredoxin,thioredoxin reductase, |
出版年 : | 2013 |
學位: | 碩士 |
摘要: | 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。 NADPH-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. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/59056 |
全文授權: | 有償授權 |
顯示於系所單位: | 農業化學系 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-102-1.pdf 目前未授權公開取用 | 2.28 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。