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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王俊能(Chun-Neng Wang) | |
| dc.contributor.author | Tzu-Huan Hong | en |
| dc.contributor.author | 洪子桓 | zh_TW |
| dc.date.accessioned | 2021-05-20T21:21:05Z | - |
| dc.date.available | 2010-10-22 | |
| dc.date.available | 2021-05-20T21:21:05Z | - |
| dc.date.copyright | 2010-10-22 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-10-20 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/10332 | - |
| dc.description.abstract | 本研究目的為在大岩桐建立可用之病毒誘導基因靜默(VIGS)系統。大岩桐為研究花部對稱性發育機制的良好材料，野生的大岩桐族群花部多為兩側對稱，但栽培種的大岩桐花部卻為輻射對稱，此對稱性之轉變可能是人為馴化過程中所產生的單一基因突變所造成，因此我們希望在大岩桐建立近年來發展出能高效率研究基因功能之VIGS系統以研究花對稱性，同時可作為其他近緣植物建立VIGS系統之參考。本研究挑選兩種可感染分生組織的VIGS病毒系統：Tobacco rattle virus (TRV)及Cymbisium mosaic virus (CymMV)嘗試在大岩桐靜默phytoene desaturase (PDS)基因。結果發現TRV雖可系統性感染大岩桐，但用攜帶大岩桐PDS的TRV接種的大岩桐PDS表現量並無顯著的下降。而用攜帶大岩桐PDS的CymMV接種的大岩桐，有些植株的PDS表現量有顯著的下降，顯示應有引發PDS靜默，但植株葉片並無出現PDS表現量下降應有的白化現象。本研究亦測試農桿菌接種，欲提高VIGS之效率，但結果顯示農桿菌對大岩桐的感染效率並不佳，因而也無法有效將病毒載體帶入植物引發VIGS。綜合本研究結果，TRV在大岩桐尚無法引發VIGS，推測是由於TRV在大岩桐無法累積足夠的病毒量引發強烈的VIGS。除了繼續改良接種的條件外，利用原本即可感染大岩桐的病毒或利用馴化得到感染力高的病毒clone重新建立VIGS載體應為未來可以考慮的方向。而CymMV雖似乎有成功引發靜默的例子但性狀並未如預期改變。為證實CymMV 可有效在大岩桐引發基因靜默並造成性狀改變，或應測試更多的報導基因。 | zh_TW |
| dc.description.abstract | This study aims to build a virus-induced gene silencing (VIGS) system in Sinningia speciosa. Sinningia speciosa, Gesneriaceae, is an ornamental plant and became an emerging model for studying flower development. In wild, flowers of Sinningia speciosa are zygomorphic, but many of their domestic conterparts are actinomorphy. The zygomorphic wild type and actinomorphic peloria provide us an excellent opportunity to compare their flower development and explore functions of candidate genes in floral symmetric regulation. VIGS is a powerful tool for studying gene functions. We chose two viral systems: Tobacco rattle virus (TRV) and Cymbidium mosaic virus (CymMV) to test their silencing ability on phytoene desaturase (PDS) in S. speciosa. Our results showed that S. speciosa could be systemically infected by TRV, but no down-regulation of PDS observed; however, CymMV caused down-regulation of PDS in some S. speciosa individuals though they did not display expected photo-bleaching. Agrobacterium inoculation was also tested. However, Agrobacterium showed poor infectivity to S. speciosa and Agrobacterium inoculation failed to help increase the VIGS efficiency. Concluding our results, the failure to induce VIGS by TRV in S. speciosa may be due to the poor infectivity. Besides optimizing the inoculation conditions, constructing new VIGS vectors from viruses that naturally occurring in S. speciosa or virus clones with more infectivity by domestication is preferred in the future. In addition, PDS of S. speciosa seems to be successfully down-regulated by CymMV, but the phenotype was not changed. To better confirm that gene silencing can be efficiently induced by CymMV in S. speciosa, more reporter genes should be tested. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-20T21:21:05Z (GMT). No. of bitstreams: 1 ntu-99-R97b44024-1.pdf: 3823128 bytes, checksum: 552b951f516ee8b41e6475615b279239 (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | 誌謝………………………..………………………………………………...i
中文摘要………………………………………………………………….....ii 英文摘要……………………………………………………………………iii 目錄………………………………………………………………………….v 圖目錄………………………………………………………………..……vii 表目錄………………………………………………………………………ix 第一章、前言……………………………………………………………….1 苦苣苔科簡介………………………………………………………………..1 大岩桐簡介…………………………………………………………………..1 病毒誘導基因靜默(VIGS) ……………………………………………………3 病毒載體系統………………………………………………………………..5 VIGS報導基因……………………………………………………..…….…..7 農桿菌媒介病毒接種…………...…………………………………..….……..7 研究目的………………………...………………………………...…...……9 第二章、材料與方法……………………………………………………...10 一、植物材料及栽培環境……………………………………..………..…….10 二、一般分生操作……………………………………..……………….……12 三、病毒載體及其衍生載體建構……………………………………..….……13 四、病毒感染力分析……………………………………..….………………17 五、農桿菌暫時性表現分析及農桿菌接種………………………………..…..18 六、fluridone離層酸抑制劑 (ABA inhibitor)處理…………………..…………21 第三章、結果……………………………………………………………...22 一、病毒感染力分析……………………………………..….……………….22 二、PDS 靜默……………………………………..….…………………….31 三、農桿菌暫時性表現分析……………………………………...….……….44 四、農桿菌媒介VIGS……………………………………..….……………..51 第四章、討論……………………………………………………………...53 大岩桐的系統葉可偵測到TRV，但並未引發VIGS……….……………………53 CymMV有可能成功在大岩桐引發VIGS，但沒有產生白化性狀………………..54 若欲用TRV和CymMV系統研究大岩桐花部基因功能需考慮花梗接種……….56 農桿菌接種並非適合大岩桐VIGS系統的接種方式…………………………...56 第五章、未來展望………………………………………………………...59 對影響病毒及農桿菌感染效率的其他因子進行優化………………………..…59 建立新的VIGS病毒載體…………………………………………………….61 第六章、參考資料………………………………………………………...63 第七章、附錄……………………………………………………..……….74 | |
| dc.language.iso | zh-TW | |
| dc.title | 大岩桐病毒誘導基因靜默系統之建立 | zh_TW |
| dc.title | Establishing virus-induced gene silencing system in
Sinningia speciosa (Gesneriaceae) | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 99-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 陳仁治(Jen-Chih Chen) | |
| dc.contributor.oralexamcommittee | 葉信宏(Hsin-Hung Yeh),鄭秋萍(Chiu-Ping Cheng) | |
| dc.subject.keyword | 大岩桐,病毒誘導基因靜默,農桿菌接種, | zh_TW |
| dc.subject.keyword | Sinningia speciosa,virus-induced gene silencing,Tobacco rattle virus,Cymbisium mosaic virus,phytoene desaturas,Agrobacterium inoculation, | en |
| dc.relation.page | 88 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2010-10-20 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生態學與演化生物學研究所 | zh_TW |
| 顯示於系所單位： | 生態學與演化生物學研究所 | |
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