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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 葉汀峰 | |
| dc.contributor.author | Li-Yuan Liu | en |
| dc.contributor.author | 劉立元 | zh_TW |
| dc.date.accessioned | 2021-06-15T13:31:18Z | - |
| dc.date.available | 2019-02-24 | |
| dc.date.copyright | 2016-02-24 | |
| dc.date.issued | 2016 | |
| dc.date.submitted | 2016-02-03 | |
| dc.identifier.citation | 王仁、陳財輝、劉瓊霦（2010）臺灣長期忽視的生物資源－竹。臺灣林業期刊 36：12-18。
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/51349 | - |
| dc.description.abstract | 麻竹（Dendrocalamus latiflorus, Dla）是臺灣分布範圍最廣的竹類，生長速度快，單一生長季高度可達25 m。在麻竹稈生長過程中，細胞壁會快速累積木質素（Lignin）。麻竹稈細胞壁中具有可觀的p-Coumaric acid及Ferulic acid；而麻竹稈木質素為H-G-S type lignin，其中S lignin隨著生長而累積的量最為顯著。被子植物（Angiosperm）中S lignin主要由Caffeic acid O-methyltransferases （COMTs）進行5-O-Methylation催化生成。從麻竹稈選殖出6條DlaCOMTs，DlaCOMTs全長為1083 bp，可轉譯（Translation）出361個胺基酸。而完整DlaCOMTs皆具有COMT蛋白的特殊結構。由演化樹狀圖可知DlaCOMTs親源關係與孟宗竹（Phyllostachys pubescens）最為相近。利用qRT-PCR測得DlaCOMTs的組織特異性，在木質化程度高之麻竹稈下部的表現量最高。體外表現並純化出的DlaCOMTs重組蛋白，利用酵素動力學測得對5-HCAld （5-Hydroxyconiferaldehyde）在in vitro狀態下專一性最高，同時對5-HFA（5-Hydroxyferulic acid）為競爭型抑制。表示DlaCOMTs在整個木質素生合成途徑中，偏好走途徑下方5-HCAld催化生成SAld（Sinapaldehyde），顯示麻竹S lignin甲基化生合成途徑可能與雙子葉被子植物的S lignin生合成途徑相似。 | zh_TW |
| dc.description.abstract | Ma bamboo (Dendrocalamus latiflorus, Dla) is the most widely distributed bamboo species in Taiwan. It grows extremely fast and can reach about 25 meter in height within one growing season. During its growth, ma bamboo’s cell walls accumulate lignin rapidly. There is a considerable amount of p-coumaric acid and ferulic acid in ma bamboo’s cell walls. And ma bamboo’s lignin is a H-G-S type lignin, and it accumulates S lignin rapidly during bamboo’s growth. The 5-O-methylation of angiosperm S lignin had long been proposed to be catalyzed by the caffeic acid O-methyltransferases (COMTs). 6 DlaCOMTs were further cloned from ma bamboo culm. The full length of DlaCOMTs is composed of a 1083 bp open reading frame encoding 361 amino acids with the highest similarity to that of Phyllostachys pubescens. All DlaCOMTs have the specific protein structures of COMT family. Gene expression profiles revealed by qRT-PCR indicated that DlaCOMTs had tissue specificity and expressed in the lignification stage during culm development. Enzyme activity assays also revealed that DlaCOMTs have in vitro substrate preference for 5-HCAld (5-Hydroxyconiferaldehyde) of the monolignol biosynthetic pathway. The O-methylation activities of DlaCOMTs on 5-HFA (5-Hydroxyferulic acid) were strongly inhibited by 5-HCAld (competitive inhibition). The results implicated that DlaCOMTs would catalyze 5-HCAld to SAld (Sinapaldehyde), and the biosynthesis pathway of S lignin methylation in ma bamboo might be similar to that in dicotyledons. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T13:31:18Z (GMT). No. of bitstreams: 1 ntu-105-R01625002-1.pdf: 4369085 bytes, checksum: e8899e146b3185cbdf13c15b7568f93c (MD5) Previous issue date: 2016 | en |
| dc.description.tableofcontents | 謝誌………………………………………………………………………………...…..I
摘要………………………………………………………………………………...…II Abstract………………………………………………………………………………III 目錄……………………………………………………………….………………….IV 圖目錄………………………………………………………………………….…...VII 表目錄………………………………………………………………………..………XI I. 前言………………………………………………………………………...….1 II. 文獻回顧………………………………………………………………………3 1. 麻竹的分布與應用……………………………………………………..…3 2. 木質素在植物生長所扮演之角色……………………………………..…4 3. COMT酵素於木質素生合成之角色與功能……………………………..7 4. COMT蛋白質結構…………………………………………………..…..12 5. 調控COMT對於改良木質素生合成的影響…………………….………16 6. COMT改良木質素之應用………………………………………………21 III. 研究目的………………………..……………………………………………24 IV. 材料與方法……………………………………………………………..……25 1. 試驗材料…………………………………………………………………25 2. 麻竹稈不同生長階段之木質素染色觀察………………………………27 2.1 試材製備…………………………………………………………..…27 2.2 切片製作…………………………………………………………..…27 2.3 木質素染色…………………………………………………..………27 3. 麻竹稈不同生長階段之化學組成分分析………………………………28 3.1 木質素含量測定…………………………………………………..…28 3.2 醣類含量測定……………………………………………………..…29 3.3 醣醛酸含量測定…………………………………………………..…29 3.4 木質素單體分析…………………………………………………..…30 4. Total RNA的萃取及cDNA的製備………………………………………31 5. 麻竹COMT基因選殖………………………………………….…………31 5.1 選殖載體（Cloning vector）…………………………………………..31 5.2 轉殖菌株…………………………………………………..…………31 5.3 設計引子（Primer）…………………………………………...………32 5.4 聚合酶連鎖反應……………………………………………..………32 5.5 純化目標基因片段並定序…………………………..………………33 6. 麻竹COMT序列之親源關係（Phylogenetic）分析……………………33 7. 麻竹COMT之組織特異性………………………………………….……33 8. 麻竹總蛋白萃取及COMT重組蛋白質表現及純化……………………35 8.1 麻竹總蛋白萃取………………………………...……...……………35 8.2 蛋白質表現載體（Expression vector）…………………….…………35 8.3 蛋白質表現菌株………………………………………………..……35 8.4 麻竹COMT重組蛋白質表現及純化…………...…………………..36 9. 酵素動力學所需藥品……………………………………………………36 10. 酵素動力學反應…………………………………………………………37 10.1 反應條件…………………………………………...………………37 10.2 酵素動力學常數計算……………………………...………………38 V. 結果與討論…………………………………..………………………………39 1. 麻竹稈不同生長階段之木質素染色觀察………………………………39 2. 麻竹稈不同生長階段之化學組成分分析………………………………40 2.1 醣類及醣醛酸含量測定…………………………………………..…40 2.2 木質素含量測定………………………………………………..……41 2.3 木質素單體分析…………………………………………………..…42 3. 麻竹總蛋白反應性………………………………………………...…….43 4. 麻竹COMT基因選殖……………………………………………………45 5. 麻竹COMT序列之親源關係分析…………………………………..….48 6. 麻竹COMT之組織特異性………………………..……………………..50 7. 麻竹COMT重組蛋白質表現及純化…………..……………………….53 8. 酵素動力學反應…………..…………………………….……………….54 8.1 麻竹COMT重組蛋白動力學性質…………..……………………...55 8.2 酵素抑制反應……..…………………………………………….…...58 VI. 結論…………………………………………………..………………………62 VII. 參考文獻………………………………………………………………..……64 | |
| dc.language.iso | zh-TW | |
| dc.subject | 麻竹 | zh_TW |
| dc.subject | 竹稈 | zh_TW |
| dc.subject | 咖啡酸甲氧基轉移? | zh_TW |
| dc.subject | 酵素動力學 | zh_TW |
| dc.subject | 木質素 | zh_TW |
| dc.subject | 組織特異性 | zh_TW |
| dc.subject | 竹稈 | zh_TW |
| dc.subject | 咖啡酸甲氧基轉移? | zh_TW |
| dc.subject | 麻竹 | zh_TW |
| dc.subject | 酵素動力學 | zh_TW |
| dc.subject | 木質素 | zh_TW |
| dc.subject | 組織特異性 | zh_TW |
| dc.subject | Caffeic acid O-methyltransferases | en |
| dc.subject | Bamboo culm | en |
| dc.subject | Tissue specificity | en |
| dc.subject | Lignin | en |
| dc.subject | Bamboo culm | en |
| dc.subject | Dendrocalamus latiflorus | en |
| dc.subject | Caffeic acid O-methyltransferases | en |
| dc.subject | Dendrocalamus latiflorus | en |
| dc.subject | Enzyme kinetics | en |
| dc.subject | Enzyme kinetics | en |
| dc.subject | Lignin | en |
| dc.subject | Tissue specificity | en |
| dc.title | 麻竹稈之木質素特性與其Caffeic acid O-methyltransferase酵素動力學分析 | zh_TW |
| dc.title | Lignin characterization and the enzyme kinetics of caffeic acid O-methyltransferase in Dendrocalamus latiflorus culm | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 104-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 張上鎮,許富蘭,鄭森松,孫英玄 | |
| dc.subject.keyword | 竹稈,咖啡酸甲氧基轉移?,麻竹,酵素動力學,木質素,組織特異性, | zh_TW |
| dc.subject.keyword | Bamboo culm,Caffeic acid O-methyltransferases,Dendrocalamus latiflorus,Enzyme kinetics,Lignin,Tissue specificity, | en |
| dc.relation.page | 71 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2016-02-03 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 森林環境暨資源學研究所 | zh_TW |
| 顯示於系所單位： | 森林環境暨資源學系 | |
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| ntu-105-1.pdf 未授權公開取用 | 4.27 MB | Adobe PDF |
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