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| ???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
|---|---|---|
| dc.contributor.advisor | 黃鵬林(Pung-Ling Huang) | |
| dc.contributor.author | Shang-Shin Lee | en |
| dc.contributor.author | 李盛新 | zh_TW |
| dc.date.accessioned | 2021-06-13T03:31:43Z | - |
| dc.date.available | 2007-08-01 | |
| dc.date.copyright | 2006-08-01 | |
| dc.date.issued | 2006 | |
| dc.date.submitted | 2006-07-28 | |
| dc.identifier.citation | 1. 馬溯軒. 1988. 香蕉之體胚發生與植株再生. 源一作物組織培養之應用研討會專集. P. 181-188.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32100 | - |
| dc.description.abstract | 本試驗運用RNA 干擾 (RNA interefence, RNAi)技術專一性默化香蕉乙烯生成相關酵素ACC氧化酶基因之表達，以探討該基因在果實後熟過程中所扮演的角色。同時分析Mh-ACO1及Mh-ACO2兩基因之啟動子活性，啟動子片段接於報導基因上游，構築成表達載體，利用農桿菌媒介法轉殖至阿拉伯芥 (Arabidopsis thaliana L. Columbia)，Mh-ACO1::GUS轉殖株之GUS活性，於簇生葉之葉緣根部有微量表現外，莖葉、花器中的苞片、花絲、花藥、柱頭、長角果與離層表現亦有表現， Mh-ACO1基因之表現隨著阿拉伯芥之生長發育階段不同而異；Mh-ACO2基因於阿拉伯芥任何生長發育階段並不表現。誘導試驗結果顯示，IAA、NAA、BA、GA3、ABA、JA、SA、NaCl、乙烯、光、淹水、低溫、機械傷害與乾旱可誘導Mh-ACO1基因之表現，但僅JA可誘導Mh-ACO2基因之表現。
此外，藉由農桿菌媒介法進行香蕉基因默化質體之轉殖，植株再生後，經GUS組織化學染色法與南方氏雜交分析結果顯示，已確定將基因默化質體穩定性轉殖至北蕉 (Musa spp. cv. Pei-Chiao, AAA group) ，此等植株可供日後小片段干擾RNA (siRNA) 之分析，以進一步了解對Mh-ACO基因之默化程度。 | zh_TW |
| dc.description.abstract | RNA interference (RNAi) is a potent trigger for specific gene silencing of expression in a number of organisms and is an efficient way to shut down gene expression. In this study, transformation of silencing plasmids specific for banana ACC oxidase genes has been performed. We also analysed the promoter activity of banana ACC oxidase genes, Mh-ACO1 and Mh-ACO2. The GUS expression of Mh-ACO1::GUS transgenic Arabidopsis plants was found in rosette leaves, roots, cauline leaves, flower buds, stigma , style, anther, filament, sepals, stigmatic tissue, siliques and abscission zone. The expression of Mh-ACO1 gene differed in developmental stages, and was induced by JA, SA, IAA, GA3, ABA, NaCl, ethylene, wound, drought, flood, light and cold (4℃); On the other hand, the expression of Mh-ACO2 gene was only induced by JA and ethylene, and showed no difference at developmental stages.
Besides, the gene silencing plasmids specific for Mh-ACO1 and Mh-ACO2 were stably transformed into banana (Musa spp. cv. Pei-Chiao, AAA group) by Agrobacteria-mediated transformation system. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T03:31:43Z (GMT). No. of bitstreams: 1 ntu-95-R93628133-1.pdf: 1667899 bytes, checksum: 94b224a6f41bb9b4ccb12ff105bbb213 (MD5) Previous issue date: 2006 | en |
| dc.description.tableofcontents | 壹、 前言 1
貳、 前人研究 2 一、 RNA干擾 (RNA interference, RNAi)技術及其應用 2 (一)、 轉錄後基因默化(Post-transcriptional gene silencing, PTGS) 2 (二)、 轉錄基因默化 (Transcriptional gene silencing, TGS) 2 (三)、 RNAi 之原理 4 1、 Dicer與siRNA 4 2、 RISC之作用機制 7 3、 RNA-dependent RNA Polymerase (RdRP)之功能 7 4、 系統性RNA干擾 (Systemic RNAi) 8 (四)、 RNA interference 技術之應用 8 1、 RNAi與基因功能 9 2、 RNAi與植物抗病 9 二、 乙烯與ACC 氧化酶基因 10 (一)、 乙烯與果實後熟 10 (二)、 番茄ACC氧化酶基因 11 (三)、 香蕉ACC氧化酶基因 12 (四)、 以基因轉殖技術控制乙烯生合成 13 三、 基因啟動子活性分析 14 參、 材料與方法 16 一、 試驗材料 16 (一)、 植物材料 16 1、 香蕉 16 2、 阿拉伯芥 16 (二)、 質體材料 16 二、 試驗方法 17 (一)、 質體DNA小量製備 (mini-preparation) 17 (二)、 質體DNA純化 17 (三)、 應用農桿菌媒介法進行香蕉穩定性基因轉殖 18 (四)、 應用農桿菌媒介法進行阿拉伯芥穩定性基因轉殖 18 (五)、 阿拉伯芥轉殖株之抗生素篩選 19 (六)、 香蕉擬轉殖株之分析 20 1、 GUS活性組織化學染色法 20 2、 植物基因組DNA之抽取 20 3、 核酸探針之製備與同位素標定 21 4、 南方氏雜交分析 22 (七)、 阿拉伯芥轉殖株之誘導處理 23 1、 不同植物生長調節劑之逆境處理 23 2、 機械傷害、乾旱逆境處理 23 3、 光照與黑暗處理 24 4、 鹽類逆境處理 24 5、 不同溫度處理 24 6、 乙烯處理 24 7、 GUS活性組織化學染色法 25 肆、 結果 27 一、 香蕉ACC氧化酶基因啟動子之活性分析 27 (一)、 Mh-ACO1啟動子構築質體轉殖至阿拉伯芥穩定性表現情形 27 1、 Mh-ACO1::GUS轉殖株之不同生長發育階段GUS活性表現 27 2、 以不同逆境誘導阿拉伯芥轉殖株之GUS活性表現 32 (二)、 Mh-ACO2啟動子構築質體轉殖至阿拉伯芥穩定性表現情形 42 1、 阿拉伯芥轉殖株之不同生長發育階段GUS活性表現 42 2、 以不同逆境誘導阿拉伯芥轉殖株之GUS活性表現 42 二、 香蕉ACC氧化酶基因默化質體之轉殖與分析 49 (一)、 GUS活性組織化學染色法分析 49 (二)、 南方氏雜交分析 (Southern blot analysis) 49 伍、 討論 59 一、 香蕉ACC氧化酶之基因表現 59 (一)、 組織專一性 59 (二)、 植物生長調節劑之誘導性 61 (三)、 非生物性逆境誘導性 62 二、 香蕉ACC氧化酶基因默化質體之轉殖 63 陸、 引用文獻 65 圖表目次 圖一 Mh-ACO1::GUS與Mh-ACO2::GUS之T-DNA之示意圖 26 圖二 Mh-ACO1::GUS［(A) to (D)〕與Mh-ACO2::GUS［(E) to (H)〕轉殖株之GUS活性表現情形 30 圖三、發芽後五天之Mh-ACO1::GUS與Mh-ACO2::GUS轉殖株之GUS活性表現情形 31 圖四、Mh-ACO1::GUS與Mh-ACO2::GUS轉殖株花器之GUS活性表現情形 33 圖五、Mh-ACO1::GUS 與Mh-ACO2::GUS 轉殖株長角果之GUS活性表現情形 34 圖六、發芽後五天之Mh-ACO1::GUS與Mh-ACO2::GUS轉殖株進行機械傷害處理後之GUS活性表現情形 36 圖七、發芽後五天之Mh-ACO1::GUS轉殖株進行50 μM JA (B)、100 μM SA (C)與20 μM ABA (D)誘導處理後之GUS活性表現情形 37 圖八、發芽後五天之Mh-ACO1::GUS轉殖株進行20 μM IAA (B)、20 μM NAA (C)與20 μM GA3 (D)誘導處理後之GUS活性表現情形 38 圖九、發芽後五天之Mh-ACO2::GUS轉殖株進行50 μM JA (B)、100 μM SA (C)與20 μM ABA (D)誘導處理後之GUS活性表現情形 40 圖十、發芽後五天之Mh-ACO2::GUS轉殖株進行20 μM IAA (B)、20 μM NAA (C)與20 μM GA3誘導處理後之GUS活性表現情形 41 圖十一、Mh-ACO1::GUS與Mh-ACO2::GUS轉殖株進行100 ppm 乙烯誘導處理後之GUS活性表現情形。Mh-ACO1::GUS轉殖株之全株 (A)、果莢 (B)、花器 (C)；Mh-ACO2::GUS轉殖株之全株 (D)、果莢 (E)、花器 (F) 44 圖十二、發芽後十二天之Mh-ACO1::GUS (A)與Mh-ACO2::GUS (B)轉殖株進行非生物性誘導處理後之GUS活性表現情形 45 圖十三、發芽後十二天之Mh-ACO1::GUS (A)與Mh-ACO2::GUS (B)轉殖株進行不同種類植物荷爾蒙誘導處理後之GUS活性表現情形 46 圖十四、發芽後十二天之Mh-ACO1::GUS (B)與Mh-ACO2::GUS (D)轉殖株進行60 ppm乙烯誘導處理後之GUS活性表現情形 47 圖十五、默化Mh-ACO1、Mh-ACO2基因表現之基因構築 51 圖十六、香蕉ACC氧化酶基因Mh-ACO2 默化質體之擬轉殖株篩選 52 圖十七、香蕉ACC氧化酶基因Mh-ACO2 默化質體之擬轉殖株篩選 53 圖十八、pBI121-1AnS與pBI121-2AnS香蕉擬轉殖株之GUS活性組織化學染色的結果 54 圖十九、香蕉基因默化轉殖株基因組DNA以Mh-ACO2基因為探針之南方氏雜交分析 55 圖二十、香蕉基因默化轉殖株基因組以35S啟動子基因為探針之南方氏雜交分析結果。 56 圖二十一、香蕉基因默化轉殖株基因組以GUS基因為探針之南方氏雜交分析結果。 57 圖二十二、香蕉基因默化轉殖株栽培之情形 58 表一、香蕉ACC氧化酶基因啟動子阿拉伯芥T3轉殖株遺傳分離率分析 29 表二、Mh-ACO1::GUS與Mh-ACO2::GUS於花器與果實之表現 35 表三、Mh-ACO1::GUS與Mh-ACO2::GUS基因之誘導表現 48 | |
| dc.language.iso | zh-TW | |
| dc.subject | 香蕉 | zh_TW |
| dc.subject | ACC 氧化酶 | zh_TW |
| dc.subject | 阿拉伯芥 | zh_TW |
| dc.subject | 啟動子 | zh_TW |
| dc.subject | ACC oxidase | en |
| dc.subject | banana | en |
| dc.subject | promoter | en |
| dc.subject | Arabidopsis | en |
| dc.title | 香蕉ACC氧化酶基因啟動子活性分析與默化質體之轉殖 | zh_TW |
| dc.title | Promoter Activity Analysis and Transformation of Silencing Plasmids for Banana 1-Aminocyclopropane-1-carboxylate Oxidase Genes | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 94-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 杜宜殷(Yi-Yin Do) | |
| dc.contributor.oralexamcommittee | 鄭隨和(Shui-Ho Cheng),何國傑(Kuo-Chieh Ho) | |
| dc.subject.keyword | ACC 氧化酶,香蕉,啟動子,阿拉伯芥, | zh_TW |
| dc.subject.keyword | ACC oxidase,banana,promoter,Arabidopsis, | en |
| dc.relation.page | 76 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2006-07-28 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 園藝學研究所 | zh_TW |
| Appears in Collections: | 園藝暨景觀學系 | |
Files in This Item:
| File | Size | Format | |
|---|---|---|---|
| ntu-95-1.pdf Restricted Access | 1.63 MB | Adobe PDF |
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