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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 黃鵬林(Pung-Ling Huang) | |
| dc.contributor.author | Yu-Chen Liao | en |
| dc.contributor.author | 廖育辰 | zh_TW |
| dc.date.accessioned | 2021-05-20T20:35:37Z | - |
| dc.date.available | 2011-01-01 | |
| dc.date.available | 2021-05-20T20:35:37Z | - |
| dc.date.copyright | 2008-08-04 | |
| dc.date.issued | 2008 | |
| dc.date.submitted | 2008-07-28 | |
| dc.identifier.citation | 李盛新. 2006. 香蕉 ACC 氧化酶基因啟動子活性分析與默化質體之轉殖. 國立台灣大學園藝學研究所碩士論文.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/9687 | - |
| dc.description.abstract | 香蕉 (Musa spp.) 為更年性果實，乙烯在果實後熟中扮演重要之角色，而ACC氧化酶 (ACC oxidase, ACO) 為乙烯生合成最終步驟酵素。由香蕉中選殖出兩個ACC氧化酶基因為Mh-ACO1及Mh-ACO2。分別將兩個香蕉ACC氧化酶基因，利用農桿菌媒介法轉殖以及RNA干擾 (RNA interference, RNAi) 技術，默化其在香蕉中的表現，以探討兩基因的功能。建立北蕉懸浮培養細胞系統後，進行農桿菌媒介法轉殖，將默化Mh-ACO1基因之表現載體轉殖進入北蕉細胞之中，並利用25 mg/L的G418抗生素進行初期篩選，並在體胚形成後再提高篩選濃度，經過轉殖後的北蕉細胞，於轉殖後5個月長出胚根，待發芽後可進行GUS活性組織化學染色分析以確認轉殖株。將Mh-ACO2默化擬轉殖株利用南方氏雜交分析，結果可將擬轉殖系分成三群，並且得知為多拷貝的穩定轉殖。Mh-ACO2默化轉殖株的Mh-ACO2表現量在各轉殖株系間具有不同程度之默化效果。而在花序的不同器官上，Mh-ACO2及siRNA的表現量，也是不盡相同的。創傷逆境的處理下， Mh-ACO1會提高表現，而Mh-ACO2則否，並且Mh-ACO2默化的轉殖香蕉，Mh-ACO1表現不會受到影響。Mh-ACO2默化之轉殖香蕉果實，在後熟過程中，其果皮轉色情形、乙烯之生合成與呼吸速率之增加，均有延遲的情形，可同時達到控制果實後熟及延緩果實後熟的產業利用目的。並且推測，在香蕉中果皮及果肉表現上ACC氧化酶的表現差異，應主要為Mh-ACO2所造成，而非Mh-ACO1。Mh-ACO1表現在轉殖株與非轉殖株的果皮與果肉上，具有相同的表現趨勢，而Mh-ACO2則不同。 | zh_TW |
| dc.description.abstract | Banana (Musa spp.) is a climacteric fruits. Mh-ACO1 and Mh-ACO2 genes isolated from banana (Musa spp., AAA group) encode fruit ripening related ACC oxidase (ACO), the last enzyme in the ethylene biosynthetic pathway. Agrobacterium-mediated method and RNA interference (RNAi) were used as a strategy of Mh-ACO1 and Mh-ACO2 gene inhibition to clarify their function in banana. In the Agrobacterium-mediated method, banana (Musa ‘Pei Chiao’, AAA group) suspension cells were co-cultivated with Agrobacterium and selected with 25 mg/L G418. In 5 months, the embryos developed roots. Southern blot analysis indicated that the Mh-ACO2 silenced transgenic banana lines were divided into three types and the transformed fragments were high copy insertion. The Mh-ACO2 was decreased in different lines and different organs in transgenic bananas. The siRNAs were only expressed in petal. Mh-ACO1 was increased in wounded leaves but Mh-ACO2 was not. When compared with wild type banana, the change of peel color, ethylene production and respiration rate in MAO2 silenced transgenic banana fruits was delayed. The Mh-ACO1 and Mh-ACO2 gene expression pattern in peel and pulp were different. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-20T20:35:37Z (GMT). No. of bitstreams: 1 ntu-97-R95628115-1.pdf: 3272421 bytes, checksum: cc532bfd7e520b409a490ffc235cc7a7 (MD5) Previous issue date: 2008 | en |
| dc.description.tableofcontents | 中文摘要 1
Abstract 2 壹、前言 3 貳、前人研究 4 一、 乙烯於植物之生理作用及生合成途徑 4 (一) 乙烯於植物之生理作用 4 (二) 乙烯於植物之生合成途徑 4 (三) 植物乙烯的更年性生成表現 5 (四) 香蕉果實後熟的乙烯生合成及相關基因之研究 8 二、 ACC氧化酶之特性與基因之研究 10 (一) ACC氧化酶之特性 10 (二) ACC氧化酶基因之選殖與分析 11 (三) 香蕉 (Musa spp., AAA group) ACC氧化酶基因之選殖與分析 12 (四) 香蕉ACC氧化酶之特性與分析 13 三、乙烯相關基因默化之機制及其應用 14 (一) 延緩果實後熟策略 14 (二) RNAi及應用 15 1. 基因默化 (gene silencing) 15 2. RNAi之原理 15 3. RNAi 技術之應用 17 4. 應用 RNAi 技術抑制ACC氧化酶基因之研究 18 參、材料與方法 19 一、 香蕉 Mh-ACO1基因默化轉殖 19 (一)植物材料 19 (二)質體材料 19 (三)培養基 19 (四)農桿菌質體DNA之小量製備 19 (五)農桿菌媒介法轉殖 21 二、 香蕉 Mh-ACO2基因默化轉殖株分析 22 (一)植物材料 22 (二) GUS活性組織化學染色分析 22 (三)植物基因組DNA之抽取 23 (四) 植物基因組RNA之抽取 23 (五)南方氏雜交分析 24 (六)小片段RNA北方雜交分析 25 (七)反轉錄聚合酶連鎖反應 26 (八)香蕉葉片創傷試驗 27 (九)香蕉果實後熟試驗 27 (十)香蕉果實後熟之果皮顏色指數 27 (十一) 香蕉果實呼吸率測定 27 (十二) 香蕉果實乙烯生成量測定 28 肆、結果 29 一、 香蕉 Mh-ACO1基因默化轉殖 29 (一) 北蕉懸浮細胞之建立 29 (二) 經農桿菌媒介法轉殖之北蕉細胞抗性篩選 29 (三) 農桿菌媒介法轉殖之北蕉細胞生長分化 32 二、 香蕉 Mh-ACO2基因默化轉殖株分析 35 (一) GUS活性組織化學染色分析 35 (二) 南方氏雜交分析 35 (三) Mh-ACO2默化轉殖北蕉殖株生長情形 35 (四) 不同轉殖株系間 Mh-ACO2基因表現 42 (五) siRNA表現於Mh-ACO2基因默化轉殖株 42 (六) 香蕉ACC氧化酶基因於創傷逆境之表現 42 (七) ACC氧化酶基因於Mh-ACO2默化轉殖株之不同器官表現 45 (七) Mh-ACO2默化轉殖香蕉之果實後熟表現 50 1. Mh-ACO2默化轉殖香蕉之果實自然後熟情形 50 2. Mh-ACO2默化轉殖香蕉之果實催熟情形 54 (八) ACC氧化酶基因於Mh-ACO2默化轉殖香蕉果皮與果肉之表現 64 伍、討論 69 一、 香蕉 Mh-ACO1基因默化轉殖 69 (一) 北蕉細胞篩選抗性 69 (二) 北蕉細胞的生長分化 70 二、 香蕉 Mh-ACO2基因默化轉殖株分析 70 (一) 轉殖片段於香蕉基因組中的插入情形 70 (二) Mh-ACO2基因默化轉殖香蕉生長情形 71 (三) 不同轉殖株系間 Mh-ACO2基因默化情形 71 (四) siRNA於轉殖株中之表現情形 72 (五) ACC氧化酶基因於創傷逆境下之表現 73 (六) Mh-ACO2默化轉殖香蕉延緩果實後熟之情形 73 (七) ACC氧化酶基因於果皮與果肉之表現 74 陸、結語 78 柒、參考文獻 79 | |
| dc.language.iso | zh-TW | |
| dc.title | 香蕉ACC氧化酶基因默化轉殖之研究 | zh_TW |
| dc.title | Studies on Silencing of 1-Aminocyclopropane-1-Carboxylate Oxidase Genes in Transgenic Banana | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 96-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 杜宜殷(Yi-Yin Do) | |
| dc.contributor.oralexamcommittee | 許圳塗(Chou-Tou Shii),劉祖惠(Tsu-Hwie Liu) | |
| dc.subject.keyword | 轉殖香蕉,延緩後熟,乙烯,ACC氧化酶,RNA干擾, | zh_TW |
| dc.subject.keyword | transgenic banana,ripen delayed,ethylene,acc oxidase,RNA interference, | en |
| dc.relation.page | 87 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2008-07-30 | |
| 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 |
| 顯示於系所單位： | 園藝暨景觀學系 | |
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