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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 葉開溫(Kai-Wun Yeh) | |
| dc.contributor.author | Yu-Chen Tsang | en |
| dc.contributor.author | 臧友真 | zh_TW |
| dc.date.accessioned | 2021-06-16T13:07:55Z | - |
| dc.date.available | 2018-08-07 | |
| dc.date.copyright | 2013-08-07 | |
| dc.date.issued | 2013 | |
| dc.date.submitted | 2013-08-01 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/61632 | - |
| dc.description.abstract | 前人研究指出高溫透過細胞質抗壞血酸過氧化酶 (Oncidium cytosolic ascorbate peroxidase 1，OgcytAPX1) 影響維他命C (ascorbate) 氧化還原比例 (還原/氧化)，進而誘導文心蘭萳西品系 (Oncidium Gower Ramsey) 開花。此外，在阿拉伯芥中大量表現OgcytAPX1，經過熱處理後能夠誘導miR156的目標基因SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 3 (SPL3) 表現。目前已知miR156-SPL調控模組參與阿拉伯芥花期調控，但尚不明瞭其詳細調控機制。為了解文心蘭中是否存在miR156-SPL調控模組與其位於高溫誘導開花機制中的調控角色，本研究在文心蘭中找到三條受miR156所調控的SPL基因 (OgSPLs)，並且分析比較不同生長階段文心蘭假球莖中miR156-SPL調控模組的表現。結果顯示隨著植株生育年齡增長，miR156會逐漸下降；反之OgSPLs則逐漸上升。高溫處理能誘導假球莖與花芽中的OgSPLs表現，外加維他命C則能抑制此誘導效益。然而，不論高溫處理或外加維他命C，都不會顯著影響miR156的表現。結果說明，高溫不需透過miR156，即可直接影響OgSPLs之基因表現。此外，在阿拉伯芥cytAPX1基因缺失突變株apx1中大量表現OgSPLs，可恢復apx1較野生型晚開花的表現型，顯示OgSPLs的調控開花功能作用於OgcytAPX1之下游。綜合上述結果可推論，OgSPLs在文心蘭自然發育過程中受miR156調控植物生長週期；然而，在文心蘭植株進入成熟期後，高溫藉由維他命C氧化還原比例所誘導的開花過程，則不需經由miR156即能直接促進OgSPLs表現而提早開花。 | zh_TW |
| dc.description.abstract | Previous studies have shown that high temperature-induced flowering is regulated by Oncidium cytosolic ascorbate peroxidase 1 (OgcytAPX1) through mediating the redox state of ascorbate (AsA) in Oncidium Gower Ramsey. Ecotopically overexpressing OgcytAPX1 in Arabidopsis can induce the expression of miRNA156-targeted SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 3 (SPL3) under high-temperature treatment. The miR156-SPL regulatory module has been known to regulate flowering in Arabidopsis, but its specific regulatory role on flowering is unclear. In order to dissect the function of miR156-SPL regulatory module on high temperature-induced flowering in Oncidium. We found three possible miR156-target SPL genes in Oncidium (OgSPLs). The study on expression profiling of Oncidium pseudobulbs at different growth stages indicated that miR156 steadily decreased, and OgSPLs increased as the Oncidium matures. Higher growth temperature increased OgSPLs transcription levels in both pseudobulbs and inflorescence buds, but the effect diminished after AsA application. However, the influence on miR156 under these treatments was not significant, suggesting that OgSPLs was independent of miR156 in high temperature-induced flowering mechanism. Meanwhile, ecotopically overexpressing OgSPLs in Arabidopsis mutant apx1 restored the delayed flowering phenotype compared to WT, suggesting that flowering regulation of OgSPLs was located at the downstream of OgcytAPX1. In conclusion, these results indicate that miR156 regulates phase transition through OgSPLs; however, OgSPLs act independently of age-mediated miR156 as one of the AsA redox downstream signal on the high temperature-induced flowering processes in the adult Oncidium. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T13:07:55Z (GMT). No. of bitstreams: 1 ntu-102-R00b42007-1.pdf: 8545823 bytes, checksum: 8387789c8049ac39f9ea73d7ee618584 (MD5) Previous issue date: 2013 | en |
| dc.description.tableofcontents | 口試委員審定書 i
誌謝 ii 中文摘要 iii Abstract iv 目錄 v 圖表目錄 vii 附錄目錄 viii 第一章 前言 1 第一節 文心蘭概述 1 第二節 植物的開花生理 4 第三節 MicroRNAs對植物生理之影響 8 第四節 研究目的 15 第二章 材料與方法 17 第一節 文心蘭高溫與維他命C處理 17 第二節 小分子RAN北方點墨法 (Northern blotting) 17 第三節 擴增 miR156目標基因之全長 22 第四節 利用5’ RACE 檢測目標基因經 miR156切除後所殘餘之片段 28 第五節 miR156目標基因表現量分析 32 第六節 質粒構築與基因轉殖 33 第七節 阿拉伯芥轉殖株篩選與分析 36 第三章 結果 38 第一節 文心蘭miR156的目標基因OgSPLs全長序列的分析 38 第二節 miR156-SPL調控模組在文心蘭不同生育階段的表現情形 39 第三節 miR156-SPL調控模組在文心蘭高溫處理後的表現情形 39 第四節 外加維他命C對文心蘭miR156-SPL調控模組之影響 40 第五節 miR156-SPL調控模組在維他命C缺乏阿拉伯芥突變株之作用與功能 40 第六節 榖胱甘肽與文心蘭miR156-SPL調控模組之關聯性 41 第七節 文心蘭APX與SPL對調控高溫誘導開花之關聯性 42 第四章 討論 46 第一節 文心蘭miR156與目標基因 46 第二節 文心蘭生理年齡與miR156-SPL調控模組之關聯性 47 第三節 miR156-SPL調控模組在高溫誘導開花機制中的角色 48 第四節 榖胱甘肽與文心蘭高溫誘導開花之關聯性 49 第五節 文心蘭高溫誘導開花之調控機制 50 第六節 文心蘭miR172可能扮演的生理角色 51 第七節 未來展望 52 參考文獻 53 圖一、文心蘭SPL與阿拉伯芥 AtSPL3、9、10 及水稻 OsSPL3、12胺基酸編碼序列比對 66 圖二、文心蘭SPL與阿拉伯芥及水稻SPL之親緣演化分析 67 圖三、文心蘭miR156與目標基因SPL結合位置與辨識切位 68 圖四、文心蘭SPL基因在於不同組織表現情形 69 圖五、miR156-SPL調控模組在幼年期與成年期假球莖間之表現情形 70 圖六、miR156-SPL調控模組在不同生長階段之假球莖表現情形 71 圖七、高溫處理對miR156-SPL調控模組表現情形的影響 72 圖八、高溫處理與維他命C對miR156-SPL調控模組於假球莖表現情形的影響 73 圖九、高溫處理與維他命C對文心蘭花芽之影響 74 圖十、缺乏維他命C對阿拉伯芥miR156之影響 75 圖十一、缺乏維他命C對阿拉伯芥SPL基因與開花基因之影響 76 圖十二、榖胱甘肽 (glutathione) 對文心蘭花芽之影響 77 圖十三、35S::OgSPL9-1載體構築示意圖 78 圖十四、35S::OgSPL10-1載體構築示意圖 79 圖十五、35S::OgSPL10-2載體構築示意圖 80 圖十六、阿拉伯芥轉殖株檢測 81 圖十七、阿拉伯芥轉殖株基因表現量分析 82 圖十八、生長溫度對阿拉伯芥轉殖株OgSPLsOE與rOgSPLsOE花期之影響 83 圖十九、OgSPLs對於阿拉伯芥開花基因表現量之影響 84 圖二十、APX與溫度對於AtSPLs表現量之影響 85 圖二十一、轉殖OgSPLs對回復APX突變種阿拉伯芥延遲花期之影響 86 圖二十二、miR156-SPL調控模組參與文心蘭高溫誘導開花機制之假說模型 87 附錄一、文心蘭Gower Ramsey品系親源圖譜 88 附錄二、文心蘭Gower Ramsey生活史 89 附錄三、文心蘭Gower Ramsey各組織名稱示意圖 90 附錄四、植物miRNAs生合成途徑 91 附錄五、補充維他命C對野生型阿拉伯芥、維他命C缺乏型阿拉伯芥以及一氧化氮缺乏型阿拉伯芥花期之影響 92 附錄六、榖胱甘肽對文心蘭花期與花梗延長之影響 93 附錄七、pCAMBIA 1300載體 94 附錄八、利用overlap extension polymerase chain reaction (OE-PCR) 製造site-directed mutagenesis流程示意圖 95 附表一、引子序列 96 附表二、檢索表 99 | |
| dc.language.iso | zh-TW | |
| dc.subject | SPLs基因 | zh_TW |
| dc.subject | 細胞質抗壞血酸過氧化酶 | zh_TW |
| dc.subject | 開花 | zh_TW |
| dc.subject | 高溫 | zh_TW |
| dc.subject | miR156 | zh_TW |
| dc.subject | 文心蘭 | zh_TW |
| dc.subject | high temperature | en |
| dc.subject | SPLs | en |
| dc.subject | Oncidium Gower Ramsey | en |
| dc.subject | flowering | en |
| dc.subject | miR156 | en |
| dc.subject | cytosolic ascorbate peroxidase 1 | en |
| dc.title | 文心蘭miR156-SPL (SQUAMOSA PROMOTER BINDING PROTEIN-LIKE) 調控模組於高溫誘導開花機制之功能性探討 | zh_TW |
| dc.title | Functional study of miR156-SPL (SQUAMOSA PROMOTER BINDING PROTEIN-LIKE) regulatory module involved in high temperature-induced flowering mechanism in Oncidium Gower Ramsey | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 101-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 詹明才(Ming-Tsair Chan),高景輝(Ching-Huei Kao),吳克強(Keqiang Wu),常玉強(Yuh-chyang Charng) | |
| dc.subject.keyword | 細胞質抗壞血酸過氧化酶,開花,高溫,miR156,文心蘭,SPLs基因, | zh_TW |
| dc.subject.keyword | cytosolic ascorbate peroxidase 1,flowering,high temperature,miR156,Oncidium Gower Ramsey,SPLs, | en |
| dc.relation.page | 99 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2013-08-01 | |
| 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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