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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 葉開溫(Kai-Wun Yeh) | |
| dc.contributor.author | Yao-Chung Liu | en |
| dc.contributor.author | 劉耀中 | zh_TW |
| dc.date.accessioned | 2021-06-15T13:26:36Z | - |
| dc.date.available | 2021-04-15 | |
| dc.date.copyright | 2016-04-15 | |
| dc.date.issued | 2016 | |
| dc.date.submitted | 2016-03-17 | |
| dc.identifier.citation | 台灣行政院農業委員會。http://www.coa.gov.tw/show_index.php。
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/51169 | - |
| dc.description.abstract | 台灣貴為蘭花王國，外銷切花又以文心蘭為大宗，由於外銷切花品系過於單一，要長期符合市場需求著實困難，有鑑於此，本實驗室將文心蘭phytoene synthase (OgPSY)-RNAi構築，透過農桿菌轉殖方式，孕育出了轉殖白花文心蘭。在此試驗中我們獲取了三個獨立轉殖株，分別為MF-1、MF-14以及MF-17。依照台灣現行針對基因改造作物 (GMOs) 的相關法規，必須通過基因轉殖植物「遺傳性狀調查」以及「環境生物安全評估」兩項檢測，方能申請該基因改造作物之品種權。透過本論文之研究顯示，由於選用花瓣專一表達啟動子 (Pchrc) 驅動OgPSY-RNAi構築，僅專一地降解文心蘭花辦中OgPSY之表現；HPLC結果亦顯示轉殖文心蘭花辦中類胡蘿蔔色素明顯下降；另外分析轉殖文心蘭母本兩次開花以及組培繁殖子代之花瓣，結果顯示OgPSY-RNAi構築能穩定地表現其功能；由於MF-1在組培的子代族群 (P1) 中出現了許多變異株，故進一步利用此獨立轉殖系 (MF-1) 探討光線與溫度對Pchrc活性的影響，發現兩者環境因子都可能影響OgPSY之表現，造成花色的差異。而經由Southern blot以及inverse PCR分析，瞭解基因轉殖文心蘭皆僅具有單一拷貝數，且其T-DNA兩翼之序列皆屬於非編碼DNA。透過環境生物安全相關試驗，相較於非轉殖株，轉殖文心蘭並無顯著生物相剋作用的現象發生，且並不會影響其根圈微生物相之生長。最後，本論文之研究亦可作為日後台灣基因轉殖花卉評估的先驅及準則，建立台灣基因轉殖花卉管理之完善制度。 | zh_TW |
| dc.description.abstract | Taiwan has a reputation in the agricultural modification for Oncidesa, of the orchid Kingdom, which is mainly for export as cut flowers. One of the major crisis faced by marketing Oncidesa is its dull appearance of florets and it has remained unchanged for a very long time in any aspect of colors. Currently, the Oncidesa cut flower market is facing a new demand for a new spectrum of colors. With the aid of our previous study, we have devloped a white floret Oncidesa through RNA interference technology. However, one of the major concern deemed by the government of Taiwan is that it required a genetic characterization and environmental biosafety assessment on surrounding biosphere for any genetically modificated organisms (GMOs). Therefore in this study, our plant materials were all grown in a special netted house. Additionally, we were able to characterize this transgenic white Oncidesa and define its transgenic RNAi allele (OgPSY) through genetic characterization. Furthermore, we showed that its phenotype could be sustained within two repetitive vegetative regenerations and that the transformants showed no sign of detrimental and environmental impact to the biosphere as compared with its non-transformant counterparts. Moreover, we investigated the effects of light and temperature on the transgenic Pchrc activity and found that these two environmental factors affected the potency of OgPSY RNAi, resulting in floret color variation on independent generation line of transgenic Oncidesa. Thus, this study holds valuable information in the aspect of the viability of sustaining a stable transgenic line and successfully introducing transgenic Oncidesa in the biosphere. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T13:26:36Z (GMT). No. of bitstreams: 1 ntu-105-R02b42023-1.pdf: 7912509 bytes, checksum: b5bd77c221f495bb4d3f8dded20d90f6 (MD5) Previous issue date: 2016 | en |
| dc.description.tableofcontents | 中文摘要.................................................i
Abstract................................................ii 檢索表..................................................iii 目錄.....................................................iv 圖表目錄.................................................vi 附錄圖表目錄............................................viii 第一章 前言...............................................1 1.1 基因轉殖蜜雪文心蘭概述.............................1 1.2 基因改造生物概述...................................9 1.3 研究目的.........................................13 第二章 材料與方法.........................................15 2.1 實驗架構圖.......................................15 2.2 實驗材料.........................................16 2.3 基因轉殖蜜雪文心蘭DNA檢測.........................16 2.4 基因轉殖蜜雪文心蘭胡蘿蔔素生成途徑相關基因表現量測定.18 2.5 以HPLC分析基因轉殖文心蘭花瓣類胡蘿蔔色素之含量差異..22 2.6 利用inverse PCR釣取基因轉殖蜜雪文心蘭T-DNA之flanking sequences...............................................23 第三章 結果..............................................37 3.1 基因轉殖蜜雪文心蘭遺傳特性調查.....................37 3.1.1 轉殖株之鑑定分析............................37 3.1.2 分析外源基因在轉殖株之表現部位...............38 3.1.3 外源基因在基因轉殖植株之穩定性分析............39 3.1.4 花瓣之類胡蘿蔔素生成途徑相關基因表現差異分析...39 3.1.5 花瓣中類胡蘿蔔色素含量差異分析................40 3.1.6 基因轉殖蜜雪文心蘭T-DNA插入之拷貝數分析.......41 3.1.7 基因轉殖蜜雪文心蘭插入之T-DNA兩翼序列釣取.....41 3.2 不同光度及不同溫差對基因轉殖蜜雪文心蘭花色之影響....42 3.3 基因轉殖蜜雪文心蘭生物安全試驗.....................45 3.3.1 農桿菌依附於基因轉殖蜜雪文心蘭1號組培苗之測試..45 3.3.2 基因轉殖蜜雪文心蘭1號之生物相剋作用測試.......45 3.3.3 基因轉殖蜜雪文心蘭1號之根圈微生物分析.........46 第四章 討論..............................................48 4.1 基因轉殖蜜雪文心蘭分子遺傳特性分析.................48 4.2 基因轉殖蜜雪文心蘭南方墨點以及T-DNA兩翼序列分析.....51 4.3 環境對白花文心蘭蜜雪1號花色之影響..................52 4.4 基因轉殖蜜雪文心蘭1號之環境生物安全試驗............54 4.5 未來展望........................................57 參考文獻................................................59 圖表....................................................70 附錄表..................................................93 附錄圖..................................................94 | |
| dc.language.iso | zh-TW | |
| dc.title | 基因轉殖白花文心蘭蜜雪之遺傳性狀及環境生物安全之研究 | zh_TW |
| dc.title | Genetic characterization and biosafety assessment of transgenic Oncidesa Gower Ramsey “Honey Snow” | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 104-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 王淑珍(Shu-Jen Wang),靳宗洛(Tsung-Luo Jinn),鄭秋萍(Chiu-Ping Cheng),陳仁治(Jen-Chih Chen) | |
| dc.subject.keyword | 文心蘭,基因改造作物,遺傳特性調查,環境生物安全評估, | zh_TW |
| dc.subject.keyword | Oncidesa,genetically modificated organisms (GMOs),genetic characterization,environmental biosafety assessment, | en |
| dc.relation.page | 101 | |
| dc.identifier.doi | 10.6342/NTU201600125 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2016-03-17 | |
| 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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