Please use this identifier to cite or link to this item:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/28237
Full metadata record
???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
---|---|---|
dc.contributor.advisor | 劉麗飛(Li-Fei Liu) | |
dc.contributor.author | Yun-Yang Chao | en |
dc.contributor.author | 趙雲洋 | zh_TW |
dc.date.accessioned | 2021-06-13T00:03:20Z | - |
dc.date.available | 2007-07-31 | |
dc.date.copyright | 2007-07-31 | |
dc.date.issued | 2007 | |
dc.date.submitted | 2007-07-31 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/28237 | - |
dc.description.abstract | Mitogen-Activate Protein Kinase (MAPKs) 是參與植物生長發育、生物與非生物訊息傳遞逆境生理之重要一員,屬多基因家族。目前對各別MAPK 之生理功能及其如何參與調控生長發育或逆境耐受性之機制仍未明瞭。因此本論文首先探討 OsMAPK3 基因可能之功能,先就其核苷酸與氨基酸序列與其他水稻MAPK 基因進行同源性比對和演化圖譜分析後,並分析 OsMAPK3 基因啟動子上之順式反應 DNA 序列,結果發現OsMAPK3 與已知參與非生物逆境調控之OsMAPK4 同屬於C 群,在核酸序列上具有69% 的相似性,但與其他 OsMAPKs 序列間的相似性僅在19% ∼ 51% 之間。此外 OsMAPK3 啟動子具有受環境、荷爾蒙及調控生長之序列,例如:DRE、HRE、CRE、ABRE、ARE及MYB等。表示OsMAPK3 可能參與諸多非生物性逆境之調控。當進一步以台農67號三葉齡水耕苗,進行不同逆境的處理,結果發現ABA 、NaCl、乾旱、H2O2 與 SA 可誘導OsMAPK3 基因之表現,但處理 paraquat、CuSO4、 CdCl2 和MJ 時則否。
為了深入瞭解OsMAPK3對水稻非生物性逆境耐受性之影響,本論文進一步以基因轉殖方法得到大量表現與抑制表現基因的轉殖水稻,並進行不同的分析。結果證實若將大量表現OsMAPK3之水稻經乾旱、高鹽、低溫和巴拉刈處理後,除葉綠素含量、Fv/Fm比值與存活率增加外,亦可提升脯胺酸含量與抗氧化酵素活性增加對逆境之耐受性。相反的,抑制表現OsMAPK3 水稻經相同處理後對逆境則不具耐受性。另外,大量表現OsMAPK3水稻在幼苗期經低溫處理後可促進分蘗,顯示轉殖水稻受低溫傷害時,可藉由增加分蘗數目以提高存活率,而轉殖水稻生長至成熟期時則可發現高節位分蘗現象,此高節位分蘗芽以插枝方式種植仍可正常生長抽穗,或許與老化下之延長生長有關。 本論文成功地將位於訊息傳遞上游的調控基因轉入於水稻中,並能有效地提升水稻對於不同逆境及除草劑之耐受性,相對於僅轉入訊息傳遞下游標的基因的作物,此OsMAPK3轉殖水稻不僅能提升作物對不同逆境之耐受性,亦可提升作物對除草劑之耐性而有利於田間管理。未來研究可針對轉殖株藉由生物晶片或2-D gel 方法,分析轉殖與非轉殖水稻間有那些基因或蛋白質,因大量或抑制OsMAPK3基因之表現而受影響,以此進一步分析OsMAPK3 基因在逆境下的調控模式,此外亦可探討OsMAPK3 對水稻分蘗相關基因之影響,乙烯及生長激素含量之變化,以明瞭OsMAPK3如何改變水稻之生長發育。總而言之,本論文以轉殖方式證實OsMAPK3基因參與水稻非生物逆境及除草劑耐受性之調控,並與水稻分蘗之形成有關。 | zh_TW |
dc.description.abstract | Mitogen-activated protein kinase (MAPK) is one of the important components that know to participate in regulation of plant abiotic-and boitic- stress responses and in controlling of plant growth and development. MAPK genes belong to a multiple gene family. However, the physiological function of each individual MAPK in rice and how MAPK engages in various abiotic stresses tolerance have not yet completely determined. In this study, first we addressed the putative function of OsMAPK3 by making alignment of the amino acid and DNA sequences and conducted the phylogenetic relationship analysis of OsMAPK3 with other rice MAPK gene. Then to understand gene expression pattern of OsMAPK3, the OsMAPK3 promoter sequence was further analysis for identification of different cis-acting DNA elements. The result showed that OsMAPK3 and OsMAPK4 that already demonstrated to involve in plant abiotic stress tolerance are in the same group C. Their nucleotide sequences shared 69% similarity; while OsMAPK3 only share 19% to 51% of sequence similarities with other OsMAPK genes. Besides, several stress-, hormone- responsive and development-control related cis-acting DNA elements exist in the promoter of OsMAPK3, including DRE, HRE, CRE, ABRE, ARE, and Myb factor recognition site. When TNG67 rice seedlings were treated with different abiotic stresses and OsMAPK3 gene expression was determined, OsMAPK3 can be induced by ABA, NaCl, drought, H2O2 and SA. On the other hand, there is no effect of paraquat, CuSO4, CdCl2 and methyl jasmonate treatments
To reveal the effect of OsMAPK3 on abiotic stress tolerance of rice, we took Agrobacterium-mediated gene transfer method to produce both over-expressing and knockout OsMAPK3 transgenic rice plants and its effect on the tolerance capability of rice seedlings under drought and salt stresses was examined. The result indicated that over-expression of OsMAPK3 in rice lead to increase of higher survival ratio, chlorophyll content, Fv/Fm value, proline content and antioxidant activity under stress. This result provided conceivable functional evidence that over- expression of OsMAPK3 gene could improve the abiotic stress and herbicide tolerance. On the contrast, the OsMAPK3 knockout transgenic rice displayed intolerant phenotype against stresses. Moreover, the over-expression OsMAPK3 transgenic rice can promote new tiller formation under cold treatment and ends with new tiller at higher internodes position when reaching maturity. We concluded that these phenomena of new tiller formation may be related to the escape of cold damage and enhance survival of rice seedlings or delay the senescence to death of rice. In this thesis, we successfully transduced OsMAPK3 gene into rice and efficiently rise up the abiotic stress and herbicide tolerance. It will be worthy to note that compared to improve plant stress tolerance through transferring the signal transduction downstream gene, our approach seems better and benefit field management than traditional study. In future, we hope to apply microarray and 2-D gel analysis techniques in characterization the gene and protein expression profiles between wild type and OsMAPK3 transgenic rice plants to understand the regulation mechanism of OsMAPK3 gene under stresses. In addition, we are dedicated to elucidate the effect of OsMAPK3 on branching (tiller)-formation related genes in rice as well as change the endogenous content of ethylene and auxin that lead to alteration of rice growth and development. Taken together, by transgenic approach this study demonstrated that OsMAPK3 gene can play an important role in regulation of abiotic stress and herbicide tolerance, also affect the tiller formation in rice. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T00:03:20Z (GMT). No. of bitstreams: 1 ntu-96-D87621103-1.pdf: 6741002 bytes, checksum: 10c9b75751a0e1120e1a248b0c627496 (MD5) Previous issue date: 2007 | en |
dc.description.tableofcontents | 目錄
口試委員審定書……………………………………… i 致 謝…………………………………………….. ii 圖表目錄……………………………………………………… ix 縮寫字對照……………………………………………………… xii 中文摘要……………………………………………………… xvi 英文摘要……………………………………………………… xviii 第一章 總論 壹、前言……………………………………….…….…….. 2 貳、研究背景…………………………………….…….…….. 3 一、非生物性逆境對作物產量及品質的影響……………... 3 二、氧化逆境對植物生長發育的影響…………………….… 4 三、植物非生物性逆境耐受性的分子機制……………… 6 四、利用轉殖方式提昇植物逆境耐受性的研究.………….. 7 五、Mitogen-Activated Protein Kinase (MAPK)與植物逆境耐受性 及生長發育之關係..…………............ 9 1. MAPKs訊息傳遞的方式及組成……..……….... 9 2. 植物 MAPKs 的分類……………………………... 10 2.1 MAPKKKs 基因………………………………………….... 10 2.2 MAPKKs 基因…………………………………...... 11 2.3 MAPKs 基因………………………………………….... 12 3. MAPKs在植物體內的功能……………………….. 13 3.1 參與有絲分裂後期細胞板的建立………………….... 13 3.2 藉由賀爾蒙調控細胞生長發育…………………….... 14 3.3 傳遞外界生物性逆境刺激的訊息……….............. 15 3.4 傳遞外界非生物性逆境刺激的訊息…………………… 16 參、水稻OsMAPK3 基因生理功能之預測…….……………… 18 1. 水稻OsMAPK3 基因核酸序列之比對及演化親源譜系分 析…................................... 20 2. OsMAPK3 基因啟動子之DNA序列分析….......... 21 肆、本論文研究方向與實驗架構.…………………......... 22 伍、參考文獻…………….………….................... 24 陸、圖 表.………………………………….......... 40 第二章 大量表現OsMAPK3水稻可藉由增加抗氧化酵素活性及脯胺酸而 提高水稻對於乾旱與高鹽的耐受性 壹、中文摘要……………………..………………………… 52 貳、英文摘要…………………………………..………..... 53 參、文獻回顧…………………………………..………..... 55 肆、材料與方法…………………………………..………. 57 伍、結 果……………………………………………………… 80 1. 台農67水稻非生物逆境耐受性篩選條件之建立…. 80 2. 水稻經逆境處理後OsMAPK3基因的表現情形....... 80 3. 轉殖水稻之獲得及分子鑑定…………………..…. 81 3.1. ABA誘導OsMAPK3轉殖水稻之分子鑑定….……. 81 3.2. 水稻在乾旱與鹽逆境下OsMAPK3的基因表現……… 81 3.3. 水稻在乾旱與鹽逆境下OsMAPK3的活性表現…………… 82 4. 轉殖水稻於逆境下之生理分析…………………… 83 4.1. ABA誘導OsMAPK3轉殖水稻之耐旱性分析……. 83 4.2. ABA誘導OsMAPK3轉殖水稻之耐鹽性分析………. 83 5. ABA誘導OsMAPK3轉殖水稻經乾旱與鹽處理後脯胺酸與蔗糖 含量之變化………………………………………… 84 6. ABA誘導OsMAPK3轉殖水稻經乾旱與鹽處理後H2O2與抗氧 化酵素活性之變化…………………………………… 84 7. 轉殖水稻在鹽逆境下OsAPXs基因的表現…………… 85 陸、討 論……………………..…………................ 86 1. 轉殖OsMAPK3水稻可提高OsMAPK3基因表現與OsMAPK3 活性………………..…………............. 86 2. 轉殖OsMAPK3水稻會提高對乾旱與鹽害的耐受性… 86 3. 轉殖OsMAPK3水稻可提高脯胺酸的含量........... 87 4. 轉殖OsMAPK3水稻可提高抗氧化酵素的活性....... 88 柒、參考文獻……………………………..…………......... 90 捌、圖 表.…………………………................... 94 第三章 抑制OsMAPK3基因表現會降低水稻對高鹽與乾旱的耐受性 壹、中文摘要……………………………………….... 110 貳、英文摘要…………………..……………………....... 111 參、文獻回顧………………..………………………....... 112 肆、材料與方法……………..…………………….………… 114 伍、結 果………………………………………………….. 118 1. 不同逆境下OsMAPK3基因的表現…………………. 118 2. 轉殖水稻之分子鑑定………………………………. 118 3. 轉殖OsMAPK3-RNAi水稻之農藝性狀及生理表現… 119 4. 抗氧化酵素活性的變化…………..…………. 120 陸、討 論……………………………………....... 121 1. 乾旱、NaCl與ABA可誘導水稻內生OsMAPK3基因之表現… 121 2. 轉殖OsMAPK3-RNAi水稻會減少株高及結實率…… 121 3. 轉殖OsMAPK3-RNAi水稻會降低對乾旱與鹽害的耐受 性…….............................. 123 4. 轉殖OsMAPK3-RNAi水稻會影響抗氧化酵素的活性…...123 柒、參考文獻…………………..………………............. 125 捌、圖 表.……………………………................... 128 第四章 離層酸誘導之OsMAPK3水稻轉殖株可提昇巴拉刈之抗性 壹、中文摘要…………………………………………………… 138 貳、英文摘要…………………………………..………….... 139 參、文獻回顧……………………………..……………….... 140 肆、材料與方法…………………………..………………… 143 伍、結 果………………………………………………… 145 1. OsMAPK3基因對巴拉刈、H2O2及ABA之反應…….. 145 2. 轉殖與非轉殖水稻於ABA或巴拉刈處理下OsMAPK3之表現分 析.............……………..………….. 145 3. 轉殖與非轉殖水稻於巴拉刈處理之抗氧化酵素活性分 析…..............................….… 146 4. 轉殖OsMAPK3水稻處理巴拉刈後之抗性生理分 析..........................………….... 147 陸、討 論……………………………………………........ 148 1. H2O2與ABA而非巴拉刈可誘導水稻內生OsMAPK3基因 之表現………………………………………………………. 148 2. ABA可有效誘導轉殖水稻OsMAPK3之表現……….. 149 3. 轉殖OsMAPK3水稻經巴拉刈處理後可增加抗氧化酵素之活性 並提升對巴拉刈之抗性……………………………………. 150 4. 轉殖OsMAPK3水稻是否透過ABA 調控之相關途徑提昇抗氧 化酵素活性………………………………………………… 150 5. OsMAPK3轉殖水稻於抗除草劑之未來應用………… 151 柒、參考文獻…………………………………………........ 153 捌、圖 表.…………………………………………......... 156 第五章 大量表現OsMAPK3水稻會影響分蘗形成之節位 壹、中文摘要……………………………………………………162 貳、英文摘要……………………………………………..... 163 參、文獻回顧…………………………………………...... 165 肆、材料與方法…………………………………..………… 168 伍、結 果……………………………………………………. 170 1. 低溫處理大量表現OsMAPK 3轉殖水稻對分蘗形成的影 響….................................… 170 1.1. 不同葉齡與低溫處理的時間對轉殖水稻分蘗的影. 170 1.2. 轉殖水稻經低溫處理後分蘗形成的位置………. 170 1.3. 觀察OsTB1在轉殖OsMAPK3水稻中的變化………… 171 2. OsMAPK3轉殖水稻可形成高節位分蘗……....... 171 陸、討 論……………………..…………................. 172 1. 低溫處理OsMAPK 3轉殖水稻會影響分蘗之形成… 172 2. 轉殖OsMAPK3水稻於成熟期形成高節位之分蘗.... 172 3. 轉殖OsMAPK3 水稻與其他分蘗基因間的關係...... 173 柒、參考文獻…………………………………............. 175 捌、圖 表.…………………………………................ 177 第六章 綜合討論與未來研究方向 壹、綜合討論………………………………………………… 184 1. OsMAPK3基因於水稻不同非生物性逆境下所扮演的角色.184 2. OsMAPK3 轉殖株與其他轉殖作物對逆境與除草劑耐受 性之比較……………………………………….… 185 3. OsMAPK3於水稻非生物性逆境、除草劑耐受性與水稻分蘗節 位形成的可能模………………………………… 186 貳、未來工作與展望 1. 以Transcriptomic 研究方式探討受OsMAPK3所調控或互 動之基因…………………………………………… 188 2. 以Proteiomic 研究方式將能與 OsMAPK3 結合或互補作用 之蛋白質………………………………………… 188 3. OsMAPK3基因與其他逆境生理之關連……….…… 188 參、參考文獻…………………………………………......... 189 伍、圖 表.………………………………………............ 192 附 錄………...……………………………………………...193 | |
dc.language.iso | zh-TW | |
dc.title | 水稻OsMAPK3基因於非生物性逆境及除草劑耐性之功能研究 | zh_TW |
dc.title | Functional Characterization of OsMAPK3 Gene from Rice (Oryza sativa L.) Required for Abiotic Stress and Herbicide Tolerance | en |
dc.type | Thesis | |
dc.date.schoolyear | 95-2 | |
dc.description.degree | 博士 | |
dc.contributor.coadvisor | 張孟基(Men-Chi Chang) | |
dc.contributor.oralexamcommittee | 蘇睿智(Ruey-Chih Su),王淑珍(Shu-Jen Wang),朱鈞(Chun Chu),侯新龍(Shin-Lon Ho) | |
dc.subject.keyword | 非生物性逆境,除草劑,水稻,過度表現,RNA干擾,葉綠素螢光,脯胺酸,抗氧化酵素,分蘗(分枝), | zh_TW |
dc.subject.keyword | abiotic stresses,herbicide,rice (Orzya sativa L.),overexpression,RNA interference,chlorophyll fluorescence,proline,antioxidative enzyme,tilling (branching), | en |
dc.relation.page | 199 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2007-07-31 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 農藝學研究所 | zh_TW |
Appears in Collections: | 農藝學系 |
Files in This Item:
File | Size | Format | |
---|---|---|---|
ntu-96-1.pdf Restricted Access | 6.58 MB | Adobe PDF |
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.