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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 黃鵬林(Pung-Ling Huang) | |
| dc.contributor.author | Da-Ling Huang | en |
| dc.contributor.author | 黃大玲 | zh_TW |
| dc.date.accessioned | 2021-06-08T04:16:48Z | - |
| dc.date.copyright | 2010-08-11 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-07-30 | |
| dc.identifier.citation | 1. Aharoni A., C. H. R. De Vos, M. Wein, Z. Sun , R. Greco, A. Kroon, J. N. M. Mol and A. P. O'Connell. 2001. The strawberry FaMYB1 transcription factor suppresses anthocyanin and flavonol accumulation in transgenic tobacco. Plant J. 28:319-332.
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Interlinking showy traits: co-engineering of scent and colour biosynthesis in flowers. Plant Biotechnol. J. 6:403–415. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/22391 | - |
| dc.description.abstract | 植物 MYB 家族基因扮演轉錄因子的角色，其功能包括調控次級代謝、細胞形態發生、花朵香氣及顏色等。本試驗選殖蝴蝶蘭 MYB cDNA 並分析其特性和功能，期望未來應用於蝴蝶蘭等花卉之育種改良。利用阿拉伯芥 Production of Anthocyanin pigment 1 (PAP1) cDNA為探針，以溶斑雜交法篩選蝴蝶蘭 cDNA 庫一百五十萬個溶斑形成單位 (plaque forming unit, pfu)。篩選出三個可演繹出完整胺基酸序列之 MYB cDNA，命名為 PtMYB1、PtMYB2 和 PtMYB3。將其胺基酸序列與資料庫序列比對後，皆屬於 R2R3-MYB 轉錄因子，含有為兩個重覆 MYB 蛋白功能性組區，每一個功能性組區皆含有三個 α - helices。為了解 PtMYB 於蝴蝶蘭基因組分佈情形，南方氏雜交分析得知三個蝴蝶蘭 PtMYB 皆為低拷貝基因。分析蝴蝶蘭不同器官 PtMYB 之表現情形，結果 PtMYB1 和 PtMYB2 於花器表現量最高，PtMYB3 於根部表現量最高。而於不同荷爾蒙誘導處理條件下，PtMYB1 可受多種荷爾蒙誘導；PtMYB2 於水楊酸處理下，表現量最高；而 PtMYB3 於水楊酸和離層酸處理下，mRNA 累積量最多。於阿拉伯芥原生質體和洋蔥表皮細胞進行細胞內定位分析，結果顯示 PtMYB1 和 PtMYB2 皆表達於細胞核。 | zh_TW |
| dc.description.abstract | Functions of MYB gene family in plants are diverse including regulation of secondary metabolism, cellular morphogenesis, flower color, and fragrance etc. Plaque hybridization was employed to screen the cDNA library of Phalaenopsis for MYB cDNA using Arabidopsis thaliana PAP1 cDNA as probe. The MYB cDNAs were isolated, characterized and named PtMYB1, PtMYB2 and PtMYB3. All of them belong to R2R3-MYB subfamily containing two repeats and each repeat has a helix-turn-helix structure. Southern blot analysis indicated that all of three PtMYB were low copy genes. Northern blot analysis revealed that their expressions were tissue-specific. PtMYB1 and PtMYB2 had higher mRNA expression level in floral organs whereas PtMYB3 had higher mRNA expression level in roots. While PtMYB1 was found to be induced by several hormones, PtMYB2 and PtMYB3 were induced mainly by salicylic acid and abscisic acid. Subcellular targeting analysis in Arabidopsis protoplasts and onion epidermal cells indicated that PtMYB1 and PtMYB2 were localized in the nucleus. | en |
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| dc.description.tableofcontents | 內容目錄
中文摘要 i Abstract ii 壹、前言 1 貳、前人研究 2 一、植物MYB基因家族 2 (一) 植物MYB基因之分類和功能 2 (二) N 端保守區域決定 MYB 表達位置 2 (三) C 端胺基酸序列與 MYB 活化功能相關 3 (四) MYB 與其他轉錄因子之共同作用 3 (五) 蘭科植物之相關MYB基因 3 二、植物 R2R3-MYB 參與 phenylpropanoid pathway 之調控 4 (一) R2R3-MYB 調節花朵及果實顏色 4 (二) R2R3-MYB與花朵香氣之關係 6 三、植物 R2R3-MYB 與防禦機制之關係 7 (一) R2R3-MYB 與生物性逆境 (biotic stress) 抗性之關係 7 (二) R2R3-MYB 與非生物性逆境 (abiotic stress) 抗性之關係 8 參、材料與方法 11 一、材料 11 (一) 植物材料 11 (二) cDNA庫 11 (三) 探針 11 二、試驗方法 11 (一) 互補DNA庫之篩選及純化 11 (二) 蝴蝶蘭基因組DNA之抽取 14 (三) 南方氏雜交分析 (Southern analysis) 14 (四) 蝴蝶蘭 RNA 之抽取 15 (五) 北方雜交分析 (Northern analysis) 16 (六) 基因片段之次選殖 17 (七) GFP融合蛋白載體構築 18 (八) 質體 DNA 大量製備 19 (九) 原生質體 PEG 轉殖法 20 (十) 基因槍 (particle bombardment) 轉殖法 21 肆、結果 27 一、蝴蝶蘭 PtMYB cDNA 之選殖與分析 27 二、蝴蝶蘭 PtMYB 基因組之拷貝數分析 28 三、蝴蝶蘭 PtMYB 基因之表現分析 29 (一) 探針的專一性 29 (二) 兩品種蝴蝶蘭之 PtMYB 器官表現特異性 29 (三) 蝴蝶蘭 PtMYB 於不同植物生長調節劑處理後之基因表現 30 四、細胞間蛋白質定位分析 31 (一) 阿拉伯芥原生質體之蛋白質定位分析 31 (二) 洋蔥上皮細胞之蛋白質定位分析 31 伍、討論 56 一、蝴蝶蘭 PtMYB1、PMYB2 和 PtMYB3 cDNA 之選殖與分析 56 二、蝴蝶蘭 PtMYB1、PtMYB2 和 PtMYB3 與其他植物 MYB 之蛋白同源性低 57 三、PtMYB1、PtMYB2 和 PtMYB3 均為低拷貝基因 58 四、PtMYB1 和 PtMYB2 於花器中有最高之 mRNA 累積量 58 五、PtMYB3 主要表現於根部 59 六、PtMYB1可受多種荷爾蒙所誘導表現 60 七、PtMYB2 和 PtMYB3 可能參與 SA 抗病或非生物性逆境抗性之機制 60 八、PtMYB1 和 PtMYB2 蛋白專一表達於細胞核中 61 陸、結論 64 柒、參考文獻 65 圖表目次 圖一、抽取 RNA 之蝴蝶蘭各部位名稱。 23 圖二、蝴蝶蘭 PtMYB1、PtMYB2 與 GFP 融合蛋白質體之構築。 24 圖三、GFP 融合蛋白載體 pPtMYB1gfp 之構築示意圖。 25 圖四、GFP 融合蛋白載體 pPtMYB2gfp 之構築示意圖。 26 圖五、蝴蝶蘭 PtMYB1 互補 DNA 之核苷酸序列及胺基酸序列。 33 圖六、蝴蝶蘭 PtMYB2互補 DNA 之核苷酸序列及胺基酸序列。 34 圖七、蝴蝶蘭 PtMYB3互補 DNA 之核苷酸序列及胺基酸序列。 35 圖八、比對蝴蝶蘭 PtMYB 蛋白和其他植物之 R2R3- MYB 蛋白之胺基酸序列。 ………………………………………………………....………..39 圖九、蝴蝶蘭 PtMYB 蛋白與 其他物種 R2R3-MYB蛋白之親源分析。 ..…………………………………………………………….…….41 圖十、PtMYB1 於紅花蝴蝶蘭 (Phalaenopsis Tai Lin Redangel) 和白花紅唇朵麗蝶蘭 (Doritaenopsis Mount Lip) 之基因組拷貝數分析。 42 圖十一、PtMYB2 於紅花蝴蝶蘭 (Phalaenopsis Tai Lin Redangel) 和白花紅唇朵麗蝶蘭 (Doritaenopsis Mount Lip) 之基因組拷貝數分析。 43 圖十二、PtMYB3 於紅花蝴蝶蘭 (Phalaenopsis Tai Lin Redangel) 和白花紅唇朵麗蝶蘭 (Doritaenopsis Mount Lip) 之基因組拷貝數分析。 44 圖十三、兩個蝴蝶蘭 PtMYB cDNA 片段交互雜交分析。 45 圖十四、蝴蝶蘭 PtMYB1、PtMYB2 和 PtMYB3 基因於紅花蝴蝶蘭 (Phalaenopsis Tai Lin Redangel) 不同部位之表現。 46 圖十五、蝴蝶蘭 PtMYB1、PtMYB2 和 PtMYB3 基因於白花紅唇朵麗蝶蘭 (Doritaenopsis Mount Lip) 不同部位之表現。 47 圖十六、紅花蝴蝶蘭 (Phalaenopsis Tai Lin Redangel) PtMYB1 於不同誘導物處理葉片後之基因表現。 48 圖十七、白花紅唇朵麗蝶蘭 (Doritaenopsis Mount Lip) PtMYB1 於不同誘導物處理葉片後之基因表現。 49 圖十八、紅花蝴蝶蘭 (Phalaenopsis Tai Lin Redangel) PtMYB2 於不同誘導物處理葉片後之基因表現。 50 圖十九、白花紅唇朵麗蝶蘭 (Doritaenopsis Mount Lip) PtMYB2 於不同誘導物處理葉片後之基因表現。 51 圖二十、紅花蝴蝶蘭 (Phalaenopsis Tai Lin Redangel) PtMYB3 於不同誘導物處理葉片後之基因表現。 52 圖二十一、白花紅唇朵麗蝶蘭 (Doritaenopsis Mount Lip) PtMYB3 於不同誘導物處理葉片後之基因表現。 53 圖二十二、PtMYB1 及 PtMYB2 於阿拉伯芥原生質體進行蛋白質定位分析。 54 圖二十三、PtMYB1 及 PtMYB2 於洋蔥表皮細胞之蛋白質定位分析。 55 圖二十四、PtMYB1、PtMYB2 和 PtMYB3 之 R2R3 蛋白功能組區之胺基酸序列比對。………………… 63 表一、蝴蝶蘭 PtMYB cDNA 選殖系分群。……………………………………….32 表二、蝴蝶蘭選殖系 pPtPAP1-28 和 pPtPAP1-12 所含cDNA 閱讀框架相對應序 列之比較。 ………………………………………………………………36 表三、蝴蝶蘭選殖系 pPtPAP1-32 和 pPtPAP1-15所含 cDNA 閱讀框架相對應序列之比較。 ………………………………………………………………37 表四、蝴蝶蘭 PtMYB 蛋白之演繹胺基酸分析。……………………………….38 表五、蝴蝶蘭 (PtMYB1、PtMYB2、PtMYB3)、阿拉伯芥 (AtMYB4、AtMYB9、AtMYB30、AtPAP1、AtPAP2、AtTT2)、金魚草 (AmROSEA1、AmROSEA2、AmVENOSA)、矮牽牛 (PhAN2、PhODO1)、玉米 (ZmC1、ZmP1)、蓖麻 (RcODO1) 及番茄 (LeANT1) R2R3-MYB 胺基酸序列間相似度之比較。..40 | |
| dc.language.iso | zh-TW | |
| dc.subject | 非生物性逆境 | zh_TW |
| dc.subject | R2R3-MYB 轉錄因子 | zh_TW |
| dc.subject | 細胞內定位分析 | zh_TW |
| dc.subject | 苯酚類物質生合成路徑 | zh_TW |
| dc.subject | 水楊酸抗病系統 | zh_TW |
| dc.subject | abiotic stress. | en |
| dc.subject | R2R3-MYB transcription factor | en |
| dc.subject | subcellular localization | en |
| dc.subject | phenylpropanoid pathway | en |
| dc.subject | salicylic acid defense mechanism | en |
| dc.title | 蝴蝶蘭 MYB cDNAs 之選殖與分析 | zh_TW |
| dc.title | Cloning and Analysis of MYB cDNAs from Phalaenopsis spp. | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 杜宜殷(Yi-Yin Do) | |
| dc.contributor.oralexamcommittee | 黃銓珍(Chang-Jen Huang),何國傑(Kuo-Chieh Ho) | |
| dc.subject.keyword | R2R3-MYB 轉錄因子,細胞內定位分析,苯酚類物質生合成路徑,水楊酸抗病系統,非生物性逆境, | zh_TW |
| dc.subject.keyword | R2R3-MYB transcription factor,subcellular localization,phenylpropanoid pathway,salicylic acid defense mechanism,abiotic stress., | en |
| dc.relation.page | 73 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2010-07-30 | |
| 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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