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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 鄭石通 教授(Shih-Tong Jeng) | |
dc.contributor.author | Wei-An Tsai | en |
dc.contributor.author | 蔡濰安 | zh_TW |
dc.date.accessioned | 2021-06-08T01:17:46Z | - |
dc.date.copyright | 2014-09-02 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-08-12 | |
dc.identifier.citation | Adam, H., Marguerettaz, M., Qadri, R., Adroher, B., Richaud, F., Collin, M., Thuillet, A.C., Vigouroux, Y., Laufs, P., Tregear, J.W., and Jouannic, S. (2011). Divergent expression patterns of miR164 and CUP-SHAPED COTYLEDON genes in palms and other monocots: Implication for the evolution of meristem function in angiosperms. Mol. Biol. Evol. 28: 1439-1454.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18660 | - |
dc.description.abstract | 近年研究指出,microRNA (miRNA) 在植物遭受逆境時,表現量會快速的變化以調控植物對逆境的適應性;然而,目前miRNA調控熱逆境反應的相關研究並不多。在本研究中,首先以DNA微陣列 (microarray) 分析,發現有兩群基因在熱處理下會有反應,分別為miR173以及miR164的目標基因。利用即時定量PCR與北方墨點法確認在熱逆境下,miR164表現量會上升,而miR164的目標基因CUC1、CUC2、NAC1、ANAC092與At3g12977會被抑制;相反的,miR173表現會下降,其中miR173的目標基因At4g29770和At5g18040表現情形則會與miR173相反。此外,mir164a T-DNA突變株在長時間熱逆境下會有明顯不耐熱的性狀。並且,也發現mir164 a和mir164b T-DNA突變株在高溫下,熱休克蛋白HSP21以及HSP17.6表現量都較野生型植株低。而在miR173的研究中,則是大量表現miR173植株會呈現對熱較為敏感的性狀,但是熱休克蛋白的表現與野生型植株的則沒有明顯差異。綜合實驗的結果得知,在高溫逆境下,植物缺乏miR164a時,植物中的miR164無法有效的抑制目標基因,並影響HSP21和HSP17.6的表現,使植物無法耐受熱逆境;而miR173則是在高溫下會被抑制,並以未知的方式來幫助植物抵抗熱逆境。 | zh_TW |
dc.description.abstract | MicroRNAs (miRNAs) are considered as an integral part of plant stress regulatory networks. However, limited research has been conducted on the heat-responsive miRNAs. In this study , microarray analyses were used to examine the changes of gene profile under heat stress. Two groups of genes responding to heat stress were obtained, and they were targeted by miR164 and miR173, respectively. Quantitative RT-PCR and northern blotting reveal that the amount of miR164 was accumulated when plants were treated with heat stress, whereas its target genes CUC1, CUC2, NAC1, ANAC092, and At3g12977 were repressed. However, the expression of miR173 was reduced by heat treatment, and two of its target genes, At4g29770 and At5g18040, were induced upon heat treatment. In the phenotypic analyses, mir164a T-DNA insertion lines were more sensitive to heat than wild-type plant in the long-term heat stress. In addition, the expression of HSP21 and HSP17.6 was reduced in mir164a and mir164b T-DNA lines compared to those in wild-type plant after heat treatment. Contrary to miR164 responses, miR173 overexpression lines were more sensitive to heat stress than wild-type. But, HSPs expression showed no significantly different between miR173 transgenics and wild-type. Taken together, miR164 and miR173 were involved in heat tolerance either through regulating HSPs or specific target genes in an unknown pathway. | en |
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dc.description.tableofcontents | 目錄
致謝................................................................................................................................ I 中文摘要....................................................................................................................... II 英文摘要......................................................................................................................III 縮寫對照表...................................................................................................................V 目錄..............................................................................................................................VI 圖表目錄...................................................................................................................VIII 第一章 序論........................................................................................................1 1. 熱逆境........................................................................................................1 1.1 熱逆境對植物的影響....................................................................1 1.2 植物抵抗熱逆境之反應................................................................2 2. 植物小分子RNA......................................................................................3 2.1植物小分子RNA之分類.......................................................................4 2.2 microRNA以及tasiRNA之生合成.......................................................4 2.3植物小分子RNA調控目標基因之方式...............................................5 2.4 miRNA在植物生長發育以及逆境中扮演的角色................................6 3. miR164 ....................................................................................................... 7 4. miR173 ....................................................................................................... 8 5. 研究目的與方向......................................................................................10 第二章 材料方法..............................................................................................11 1、 植物材料..................................................................................................11 2、 熱處理方式..............................................................................................11 3、 阿拉伯芥葉片組織DNA萃取...............................................................12 3.1 DNA萃取..............................................................................................12 3.2 PCR反應...............................................................................................12 4、 Total RNA萃取.......................................................................................13 5、 小分子RNA分離...................................................................................14 6、 小分子RNA電泳...................................................................................14 7、 北方墨點分析..........................................................................................14 7.1 RNA轉印..............................................................................................14 7.2探針製備................................................................................................15 7.3雜核反應與訊號分析............................................................................16 8、 質體構築..................................................................................................16 8.1 DNA黏合反應......................................................................................16 8.2轉型作用................................................................................................17 8.3植體DNA的抽取與挑選.....................................................................17 VII 9、 RT-PCR與即時定量PCR.......................................................................18 9.1反轉錄反應(Reverse transcription).....................................................18 9.2即時定量聚合酶連鎖反應....................................................................18 10、 阿拉伯芥耐熱生理活性分析..................................................................19 11、 阿拉伯芥原生質體短暫表現..................................................................19 11.1原生質體萃取......................................................................................19 11.2阿拉伯芥原生質體短暫表現目標基因..............................................20 第三章結果................................................................................................................21 1.阿拉伯芥高溫逆境反應之miRNAs調取......................................................21 2.miR164在熱逆境下表現之確認.....................................................................21 3.miR164大量表現轉殖株以及T-DNA突變株的鑑定與篩選.......................22 4. miR164各轉殖株與突變株於幼年期之耐熱表型以及分子層次分析........22 4.1下胚軸伸長測試....................................................................................22 4.2長時間熱逆境之生理分析....................................................................23 4.3分子層次調控........................................................................................24 5.miR164各轉殖株與突變株在生長後期耐熱表型測試.................................24 6.miR173在熱逆境下表現之確認.....................................................................25 7.miR173大量表現轉殖植株以及tasiRNAs生合成之T-DNA突變株之鑑定與篩選..................................................................................................................25 8. miR173大量表現植株、TAS1/TAS2生合成缺失突變株與野生型阿拉伯芥之耐熱表型及分子層次分析..............................................................................26 8.1獲得性耐熱能力之分析........................................................................26 8.2長時間非致死高溫(37oC)下發芽率之分析.......................................27 9. 未知功能之TAS1目標基因At4g29770之分析.........................................28 第四章討論................................................................................................................29 1. 高溫逆境與miRNA.......................................................................................29 2. miR164各轉殖株在前期熱逆境外表型及分子層次分析之探討................29 2.1外表型探討............................................................................................29 2.2分子層次之探討....................................................................................31 3. miR164各轉殖株在後期熱逆境外表型及分子層次分析之探討................32 4. miR173大量表現植株、野生型植株以及tasiRNAs生合成突變株(rdr6-11和dcl4-2t) 在熱逆境外表型以及分子層次之探討..........................................33 4.1獲得性耐熱反應之探討........................................................................33 4.2種子發芽率之探討................................................................................34 5. 分析TAS1目標基因At4g29770之探討..............................................34 6. 總結..........................................................................................................35 第五章參考文獻........................................................................................................51 VIII 圖表目錄 圖一. 野生型阿拉伯芥在長時間高溫35oC及常溫22oC處理下miR164和其目標基因的表現量..............................................................................................................36 圖二.miR164大量表現轉殖植株以及T-DNA突變株的鑑定與篩選.....................37 圖三.野生型和過量表現miR164及mir164 T-DNA植株在幼年生長時期的耐熱性測試..............................................................................................................................38 圖四.野生型及miR164各轉殖株的耐熱生理分析..................................................39 圖五.野生型及miR164各轉殖株在長時間熱逆境下分子層次之分析..................40 圖六.野生型及各轉殖株生長後期之耐熱表現型分析.............................................42 圖七.野生型阿拉伯芥在37oC誘導性高溫及22oC常溫處理下miR73、siR255、siR614和其目標基因的表現量.................................................................................43 圖八.miR173大量表現轉殖植株以及dcl4-2t、rdr6-11 T-DNA突變株的鑑定與篩選..................................................................................................................................44 圖九.大量表現miR173之轉植株、rdr6-11和dcl4-2t T-DNA突變株以及野生型植株之獲得性耐熱能力(acquired thermotolerance) 及分子層次分析..................45 圖十.大量表現miR173之轉植株、rdr6-11和dcl4-2t T-DNA突變株以及野生型植株在高溫下的發芽率..............................................................................................46 圖十一.At4g29770發生alternative splicing轉錄出缺乏TAS1辨識續列的mRNA ......................................................................................................................................47 圖十二.進行miR164目標基因NAC1與ANAC092之NARD domain比對........48 表一.引子序列表.........................................................................................................49 | |
dc.language.iso | zh-TW | |
dc.title | 在熱逆境下阿拉伯芥中miR164與miR173的調節功能 | zh_TW |
dc.title | Functions of miR164 and miR173 in Arabidopsis upon heat stress | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 王愛玉 教授(Ai-Yu Wang),洪傳揚 副教授(Chwan-Yang Hong),吳少傑 副教授(Shaw-Jye Wu) | |
dc.subject.keyword | miR164,miR173,長時間熱逆境,At4g29770,ANAC092, | zh_TW |
dc.subject.keyword | miR164,miR173,long-term heat stress,At4g29770,ANAC092, | en |
dc.relation.page | 62 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2014-08-12 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 植物科學研究所 | zh_TW |
顯示於系所單位: | 植物科學研究所 |
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