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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 葉開溫(Kai-Wun Yeh) | |
| dc.contributor.author | Zi-Long Yu | en |
| dc.contributor.author | 游子隆 | zh_TW |
| dc.date.accessioned | 2021-06-17T06:02:43Z | - |
| dc.date.available | 2019-01-30 | |
| dc.date.copyright | 2019-01-30 | |
| dc.date.issued | 2019 | |
| dc.date.submitted | 2019-01-29 | |
| dc.identifier.citation | 中華民國行政院農委會。http://www.coa.gov.tw/show_index.php。
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Comparison of carotenoid accumulation and biosynthetic gene expression between Valencia and Rohde Red Valencia sweet oranges. Plant Sci. 227: 28-36 Winkel-Shirley, B. (2001). Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiology 126: 485-493. Zeng, W., Huang, M., Wang, X., Ampomah-Dwamena, C., Xu, Q., and Deng, X. (2013). Identification and functional characterization of the promoter of a phytoene synthase from sweet orange (Citrus sinensis Osbeck). Plant Molecular Biology 31: 64-74. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/71536 | - |
| dc.description.abstract | 切花文心蘭 ( Oncidesa ) 為台灣切花最大宗之外銷花卉，其為目前相當重要之經濟作物。然近年切花文心蘭因品系缺乏變化，使出口總值趨於平緩。為突破此一瓶頸，本實驗室利用RNA干擾技術 ( RNA interference ) 進行基因轉殖，創造出新白花品系——蜜雪文心蘭( Oncidesa Honey Snow MF )。此基因轉殖載體之啟動子pOgCHRC ( chromoplast-specific carotenoid-associated protein ) ，為文心蘭花部組織專一表達之啟動子。研究顯示，白花文心蘭具有受光照影響花色之特性。經由光照試驗結果顯示，白花文心蘭於較高強度光照之環境下其花色偏黃之比例增加。為了解影響花色變化之因素，本論文針對參與在此RNA干擾現象內之相關因子做進一步之分析，包括：RNA干擾之目標基因OgPSY、類胡蘿蔔素生合成路徑之基因、驅動RNAi效果之啟動子pOgCHRC。結果顯示，啟動子pOgCHRC於此花色變化現象中扮演關鍵之角色。進一步以阿拉伯芥轉殖分析此啟動子之特性，發現此啟動子pOgCHRC於-1390至-1034之區域，可能存在一負調控因子，將此段序列做序列分析，顯示其上具有一段Box 4之調控因子 ( ATTAAT )，Box 4為一段與光調控有關之保守序列。而在其序列-1033至-712之區段，具有一段CGT-motif ( CCTGAACGC ) 及一段GT1-motif ( GGTTAAT )，此區段會增強啟動子pOgCHRC之活性，而高強度光照會抑制啟動子之活性。研究結果顯示pOgCHRC啟動子於-1033至-1具有最強之表達能力，因此此片段可提供未來提升文心蘭轉殖株花色品質之理想啟動子。 | zh_TW |
| dc.description.abstract | In Taiwan, Oncidesa is the top cut-flower exported to Japan. In our previous study, a new transgenic white florets “Oncidesa Honey Snow MF” was generated by silencing phytoene synthase (OgPSY). The RNAi construct is driven by a tissue-specific promoter: pOgCHRC (The promoter of chromoplast-specific carotenoid-associated protein), which is specifically expressed in floral tissues. However, some of the Oncidesa MF showed yellow or pale yellow flower. The ratio of white and yellow flowers seems to be influenced by temperature or light intensity. To realize the effects on flower colour, we analyzed the carotenoid biosynthesis gene and OgCHRC. We demonstrated that the activity of pOgCHRC is crucial for the formation of flower color. To study the regulatory mechanism of pOgCHRC, the different deleted promoter regions were cloned unto a GUS reporter gene and transformed into Arabidopsis thaliana. We identified that the promoter region from -1390 to -1034 repressed the activity of pOgCHRC. It contains a cis-regulating element: Box 4, which is predicted to be involved in light response. In the promoter region from -1033 to -712, the CGT-motif and the GT1-motif ware found, which may enhance the activity of pOgCHRC. The result shows that the promoter region from -1033 to -1 has the highest activity. In the future, the truncated promoter can be used to promote the RNA interference efficiency in Oncidesa transformants. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T06:02:43Z (GMT). No. of bitstreams: 1 ntu-108-R04b42034-1.pdf: 3729325 bytes, checksum: 696d47c0a3a2717e4e979b2fe20e4ef9 (MD5) Previous issue date: 2019 | en |
| dc.description.tableofcontents | 口試委員會審定書 i
致謝 ii 摘要 iii Abstract iv 檢索表 v 目錄 vi 圖表目錄 x 附錄目錄 xii 第一章 前言 1 1.1. 文心蘭概述 1 1.1.1. 蘭花簡介 1 1.1.2. 文心蘭簡介 1 1.1.3. 台灣文心蘭之產業與銷售現況 2 1.2. 基因轉殖白花文心蘭概述 3 1.2.1. 緣起 3 1.2.2. 文心蘭花色色素之鑑定與分析 4 1.2.3. 植物色素概述 5 1.2.4. 文心蘭基因轉殖與啟動子pOgCHRC 5 1.2.5. 環境對白花文心蘭花色之影響 7 1.3. 本論文研究目的 8 第二章 材料與方法 10 2.1. 實驗材料 10 2.1.1. 植物材料 10 2.1.2. 微生物材料 10 2.2. 文心蘭基因表現量測定 10 2.2.1. 抽取文心蘭之總體RNA 10 2.2.2. RNA膠體電泳 12 2.2.3. 合成cDNA 13 2.2.4. 反轉錄聚合酶連鎖反應 ( Reverse transcription-Polymerase Chain Reaction, RT-PCR ) 14 2.2.5. DNA 膠體電泳 15 2.2.6. 即時定量聚合酶連鎖反應 ( Real-time quantitative PCR ) 16 2.3. 分析栽培環境對文心蘭基因表現之影響 17 2.3.1. 不同光照強度與基因表現 17 2.3.2. 不同溫度與基因表現 17 2.4. 載體構築及轉型 18 2.4.1. 製備欲構築之DNA片段 18 2.4.2. DNA膠體純化 19 2.4.3. 黏合反應 ( ligation ) – TA cloning 19 2.4.4. 大腸桿菌勝任細胞之轉型 ( transformation ) 20 2.4.5. 大腸桿菌之載體檢測 21 2.4.6. 質體抽取 21 2.4.7. 限制酶切割反應 22 2.4.8. 接合反應 ( ligation ) 22 2.5. 阿拉伯芥基因轉殖與分析 23 2.5.1. 阿拉伯芥栽種 23 2.5.2. 農桿菌轉型 ( 熱穿孔轉殖法 ) 24 2.5.3. 農桿菌轉殖前處理 25 2.5.4. 阿拉伯芥之農桿菌轉殖 26 2.5.5. 阿拉伯芥之轉殖株篩選 26 2.5.6. 阿拉伯芥轉殖株之轉殖基因檢測 27 2.6. 阿拉伯芥轉殖株之GUS染色 28 2.7. 轉殖基因於阿拉伯芥轉殖株不同部位之表現量 30 2.8. 光照強度對阿拉伯芥轉殖株之影響 30 2.9. 文心蘭基因槍粒子轟擊試驗 ( Particle bombardment ) 31 第三章 結果 32 3.1. 文心蘭內生性OgCHRC基因之各組織表現量分析 32 3.2. 檸檬心文心蘭與白花文心蘭蜜雪之OgPSY基因表達量檢測 32 3.3. 環境對檸檬心文心蘭類胡蘿蔔素生合成路徑之基因與OgCHRC基因表現影響 33 3.4. 啟動子pOgCHRC序列分析 34 3.5. 不同長度片段之啟動子pOgCHRC載體構築 34 3.6. 阿拉伯芥pOgCHRC::GUS轉殖株之建立 35 3.7. 啟動子pOgCHRC全長於阿拉伯芥轉殖株各部位GUS表現情形 35 3.8. 不同長度啟動子pOgCHRC於阿拉伯芥轉殖株之表現情形 36 3.9. 光照強度對阿拉伯芥轉殖株pOgCHRC::GUS之影響 36 3.10. 啟動子pOgCHRC::GUS於文心蘭花朵中之短暫表現情形 37 第四章 討論 39 4.1. 文心蘭OgCHRC基因表現情形 39 4.2. 白花文心蘭OgPSY基因表現分析 40 4.3. 環境對文心蘭類胡蘿蔔素生合成與OgCHRC之影響 41 4.4. 啟動子pOgCHRC之序列分析 42 4.5. 啟動子pOgCHRC於阿拉伯芥轉殖株之表現情形 42 4.6. 光照強度對啟動子pOgCHRC之影響 43 4.7. 不同程度刪減啟動子pOgCHRC於文心蘭花朵之短暫表現 44 4.8. 未來展望 45 參考文獻 46 | |
| dc.language.iso | zh-TW | |
| dc.title | 南西文心蘭啟動子pOgCHRC之功能特性研究 | zh_TW |
| dc.title | Functional characterization of the promoter pOgCHRC in Oncidesa Gower Ramsey hybrid | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 107-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 謝旭亮(Hsu-Liang Hsieh),靳宗洛(Tsung-Luo Jinn),陳賢明(Hieng-Ming Ting),邱崇益(Chung-Yi Chiou) | |
| dc.subject.keyword | 文心蘭,RNA干擾技術,OgPSY,pOgCHRC, | zh_TW |
| dc.subject.keyword | Oncidesa,RNA interference,OgPSY,pOgCHRC, | en |
| dc.relation.page | 84 | |
| dc.identifier.doi | 10.6342/NTU201900271 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2019-01-29 | |
| 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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