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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 洪傳揚(HONG CHWAN-YANG) | |
| dc.contributor.author | Ping-Hung Hsieh | en |
| dc.contributor.author | 謝秉宏 | zh_TW |
| dc.date.accessioned | 2021-06-12T18:34:40Z | - |
| dc.date.available | 2012-08-03 | |
| dc.date.copyright | 2007-08-03 | |
| dc.date.issued | 2007 | |
| dc.date.submitted | 2007-07-31 | |
| dc.identifier.citation | 參考文獻
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/28033 | - |
| dc.description.abstract | 鎘及高溫逆境會嚴重限制植物的生長及產量。過去的研究顯示，常溫下 (日溫30℃/夜溫25 ℃)，鎘逆境使得耐鎘水稻TNG67的根部及葉片比不耐鎘水稻TCN1累積較高含量的ABA，而在高溫 (日溫35 ℃/夜溫30 ℃)與鎘的雙重逆境下，TNG67及TCN1則都會在葉片中累積高含量的ABA。為了瞭解鎘及高溫逆境對水稻ABA生合成的調控機制，本試驗以阿拉伯芥基因為藍本，經由生物資訊學的方法找出水稻中ABA生合成相關的基因，並分析這些基因對鎘及高溫的反應。試驗中一共找出2個ZEP基因、13個NCED基因、3個SDR基因、及5個AAO基因。半定量RT-PCR分析結果顯示，各基因家族在TNG67不同組織間多具有不同的表現量。TNG67及TCN1的葉片及根部對鎘及高溫有不同的反應。在葉片中，所有OsZEPs與OsAAOs的表現都不受鎘及高溫的影響，但一些OsNCEDs (OsNCED1、OsNCED2、OsNCED3、OsNCED5及OsCCD8b)、及OsSDRs (OsSDR2及OsSDR3)，則有差異表現。在根部中，OsZEPs (OsZEP1a及OsZEP1b)、OsNCEDs (OsNCED1、OsNCED2、OsNCED3、OsNCED5、OsNCED6、OsNCED7、OsCCD1及OsCCD8b)、OsSDR3、及OsAAOs (OsAAO3及OsAAO4)等基因，在鎘、高溫或高溫加鎘雙重逆境下，分別受到不同程度的調控。這些結果顯示，耐鎘與對鎘敏感水稻在鎘逆境或高溫逆境下，葉片及根部ABA生合成具有不同但複雜的調控機制。 | zh_TW |
| dc.description.abstract | Cadmium (Cd) and high temperature are limiting factors of plant growth and yield. Previous studies demonstrated that Cd stress increased the accumulation of ABA in Cd-tolerant cultivar TNG67 under normal temperature condition 30 ℃/25 ℃ (day /night), while no significant change was observed in Cd-sensitive cultivar TCN1 in both leaf and root tissue. On the other hand, with treatment of Cd and high temperature 35 ℃/30 ℃ (day /night), ABA was shown to highly accumulated in leaves of TCN1 and TNG67. To dissect the molecular mechanisms affecting ABA accumulation under Cd and high temperature stresses in rice seedling, this study identified ABA biosynthetic genes by bioinformatics approach, and gene expression patterns were analyzed by semi-quantitative RT-PCR. Totally, 2 ZEP genes, 13 NCED genes, 3 SDR genes, and 5 AAO genes were identified. Gene expression analysis revealed that genes of each gene family differentially expressed in different tissues. ABA biosynthetic genes showed different expression patterns to Cd and high temperature between Cd-tolerant cultivar TNG67 and Cd-sensitive cultivar TCN1. In leaf tissue, OsZEPs and OsAAOs did not response to Cd and high temperature stresses, but some genes of OsNCEDs (OsNCED1, OsNCED2, OsNCED, OsNCED5, OsCCD8b), and OsSDRs (OsSDR2 and OsSDR3) were regulated by Cd and high temperature. In root tissue, expression of OsZEPs (OsZEP1a and OsZEP1b), OsNCEDs (OsNCED1, OsNCED2, OsNCED3,OsNCED5, OsNCED6, OsNCED7, OsCCD1 and OsCCD8b), OsSDR3 and OsAAOs (OsAAO3 and OsAAO4) were regulated by Cd, high temperature, or Cd plus high temperature stresses. Taken together, our data reveals that under Cd, high temperature or Cd plus high temperature conditions, leaf or root tissue of TCN1 and TNG67 have diverse regulatory modes controlling the biosynthesis of ABA. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-12T18:34:40Z (GMT). No. of bitstreams: 1 ntu-96-R94623005-1.pdf: 2527101 bytes, checksum: c5d6c2c07bb44923ba3eb7b0d5ae9146 (MD5) Previous issue date: 2007 | en |
| dc.description.tableofcontents | 口試委員會審定書
志謝 目錄…………………………………………………………………………...I 圖表目錄………………….………………………………………………….………..III 表目錄…………………………………………………………………….……III 圖目錄…………………………………………………………………….……IV 縮寫字對照…………………………………………………………………………..VI 摘要……………………………………………………………………………..1 Abstract……………………………………………………………………………..2 前言…………………………………………………………….………………4 文獻回顧………………………………………………………………………5 一、 ABA的生理功能…………………………………………………………5 二、 ABA的生合成……………………………………………………………5 三、 ZEP基因在ABA生合成扮演的角色…………………………………...6 四、 NCED基因在ABA生合成扮演的角色…………………………………8 （一） CCD基因家族………………………………………………………8 （二） NCED基因家族 ……………………………………………………9 五、 SDR基因在ABA生合成扮演的角色………………….…….…………12 六、 AAO基因在ABA生合成扮演的角色……………….……….…………12 七、 ABA代謝基因調控與荷爾蒙對ABA含量的影響………………….....…13 八、 鎘逆境對植物的影響………….....................................................................13 九、 高溫逆境對植物的影響……….....................................................................14 十、 高溫與重金屬雙重逆境……….....................................................................15 材料與方法………………………………………………………………………16 一、 材料之種植、選取與處理……………………………………………………16 二、RNA製備……………………………………………………………17 三、 水稻中，可能的ZEP、NCED、SDR及AAO基因搜尋……………………19 四、 NCED基因可能的葉綠體導引胜 20 五、基因專一性引子對的設計………………………………………………20 六、 二步驟RT-PCR……………………………………………………………20 七、 PCR 產物瓊脂凝膠電泳分析………………………………………………22 八、 PCR產物之回收與定序……………….……………………………………22 九、 親緣演化樹之建立…………………………………………………………23 結果………………………………………………………………………………25 一、 氯化鎘及高溫逆境對TCN1及TNG67水稻幼苗生長的影響……………25 二、 生物資訊學及親緣關係分析水稻中ABA生合成基因……………………25 (一) ZEP基因部份……………………………………………………………25 (二) NCED基因部份 ………………………………….……………………25 (三) SDR基因部份……………………………………………………………26 (四) AAO基因部份……………………………………………………………26 三、NCED基因可能的葉綠體導引胜肽（putative transit peptide）預測……27 四、水稻不同組織中ABA生合成相關基因的表現………………………………27 (一) OsZEP……………………………………………..………………………27 (二) OsNCED……………………………………………………..……………27 (三) OsSDR……………………………………………………..………………27 (四) OsAAO…………………………………………………….………………27 五、 氯化鎘及高溫處理下，葉片中的ABA生合成相關基因的表現…………28 六、 氯化鎘及高溫處理下，根部的ABA生合成相關基因的表現……………28 (一) OsZEP……………………………………………………………..………28 (二) OsNCED………………………………………………………..…………28 (三) OsSDR……………………………………………………………..………29 (四) OsAAO…………………………………………………………….………29 討論………………………………………………………………………………51 參考文獻…………………………………………………………………………56 圖表目錄 表目錄 表1 水稻中可能的ZEP、NCED、SDR及AAO基因總表。………………65 表2a 半定量RT-PCR分析中所使用的OsNCED專一性引子對序列、預測擴增片段及黏合溫度。………………………………………………....66 表2b 半定量RT-PCR分析中所使用的OsZEP、OsSDR、OsAAO及控制組基因─Oshsp17.3、OsPDR9及OsActin專一性引子對序列、預測擴增片段及黏合溫度。……….………………………………………………....67 表3 水稻中NCED家族基因、CCD家族基因、玉米ZmVP14基因 (玉米的NCED基因)及阿拉伯芥AtNCED3基因間，胺基酸序列以及3’ UTR序列相似性比對 (%)。.………………………………………………....68 表4 水稻NCED及CCD基因中可能的葉綠體導引訊息胜肽（putative transit peptide）預測。…….………………………………………………....69 附表1 木村氏水耕液配方。…….……………………………………………….70 圖目錄 附圖1 ABA生合成與代謝之途徑 。…………………….………………………7 圖1 氯化鎘 (500 μM) 於 30 ℃/25 ℃ (日/夜溫) 與 35 ℃/30 ℃ 對水稻幼苗生長之影響 (24h)。.……………………………….………………30 圖2 氯化鎘 (500 μM) 於 30 ℃/25 ℃ (日/夜溫) 與 35 ℃/30 ℃ 對水稻幼苗生長之影響 (48h)。..…………………………………………….…31 圖3a 水稻OsZEP1基因和其他物種ZEP基因保守性序列的比對。……..32 圖3b 水稻OsZEP1基因有Alternative Splicing現象，其具有兩種基因結構。……………………………………………………………….…….33 圖4 OsZEP基因的親緣演化樹。……………………….…………….……….34 圖5 水稻OsNCED及OsCCD和其他物種NCED及CCD基因四個保守性Histidine的比對。……………………………………………….……..35 圖6 水稻OsNCED及OsCCD的親緣演化樹。……………………..….….36 圖7 NCED及CCD結構示意圖。…………………………….…….….….37 圖8 水稻OsSDR基因和其他物種SDR基因保守性序列的比對。.…...….38 圖9 水稻OsSDR的親緣演化樹。…………………………………….…….39 圖10 水稻OsAAO基因家族保守性區域和其他物種AAO基因的比對。......40 圖11 OsAAO基因的親緣演化樹。……….…..………………………….…….41 圖12 OsZEP1基因於水稻不同組織中的表現。.……….……………….…….42 圖13 OsNCED及OsCCD基因於水稻不同組織中的表現。………………….43 圖14 OsSDR基因於水稻不同組織中的表現。…………..……..……….…….44 圖15 OsAAO基因於水稻不同組織中的表現。…………..……………….…….45 圖16 三葉齡水耕栽培幼苗分別在30 ℃/25 ℃ (日/夜溫) 與 35 ℃/30 ℃下以氯化鎘 (500 μM)處理是24及48小時後，分別選取水稻幼苗第二葉(A)及根部(B)抽取RNA，進行OsZEP1基因分析。………….………………46 圖17 三葉齡水耕栽培幼苗分別在30 ℃/25 ℃ (日/夜溫) 與 35 ℃/30 ℃下以氯化鎘 (500 μM)處理是24及48小時後，選取水稻幼苗第二葉抽取RNA，進行OsNCED及OsCCD基因分析。…………..………….…….47 圖18 三葉齡水耕栽培幼苗分別在30 ℃/25 ℃ (日/夜溫) 與 35 ℃/30 ℃下以氯化鎘 (500 μM)處理是24及48小時後，選取水稻幼苗根部抽取RNA，進行OsNCED及OsCCD基因分析。..…………..………….…….48 圖19 三葉齡水耕栽培幼苗分別在30 ℃/25 ℃ (日/夜溫) 與 35 ℃/30 ℃下以氯化鎘 (500 μM)處理是24及48小時後，分別選取水稻幼苗第二葉(A)及根部(B)抽取RNA，進行OsSDR基因分析。.…………………..….49 圖20 三葉齡水耕栽培幼苗分別在30 ℃/25 ℃ (日/夜溫) 與 35 ℃/30 ℃下以氯化鎘 (500 μM)處理是24及48小時後，分別選取水稻幼苗第二葉(A)及根部(B)抽取RNA，進行OsAAO基因分析。…………..………….50 | |
| dc.language.iso | zh-TW | |
| dc.subject | ABA生合成相關基因 | zh_TW |
| dc.subject | 基因表現 | zh_TW |
| dc.subject | 高溫逆境 | zh_TW |
| dc.subject | 鎘逆境 | zh_TW |
| dc.subject | gene expression | en |
| dc.subject | Cadmium stress | en |
| dc.subject | ABA biosynthetic genes | en |
| dc.subject | high temperature stress | en |
| dc.title | 鎘及高溫調控水稻幼苗ABA生合成分子機制之研究 | zh_TW |
| dc.title | Studies on the molecular mechanism of ABA biosynthesis in response to cadmium and high temperature in rice seedlings | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 95-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 高景輝,鍾仁賜,張孟基,葉靖輝 | |
| dc.subject.keyword | ABA生合成相關基因,鎘逆境,高溫逆境,基因表現, | zh_TW |
| dc.subject.keyword | ABA biosynthetic genes,Cadmium stress,high temperature stress,gene expression, | en |
| dc.relation.page | 64 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2007-08-01 | |
| 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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