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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張英? | |
| dc.contributor.author | Yun-Jhih Shih | en |
| dc.contributor.author | 施蘊芷 | zh_TW |
| dc.date.accessioned | 2021-06-17T06:00:00Z | - |
| dc.date.available | 2021-02-14 | |
| dc.date.copyright | 2019-02-14 | |
| dc.date.issued | 2019 | |
| dc.date.submitted | 2019-02-12 | |
| dc.identifier.citation | Aitken, A. (2006). 14-3-3 proteins: a historic overview. Semin Cancer Biol 16, 162-172.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/71395 | - |
| dc.description.abstract | 蔗糖是植物養分運輸的主要形式，而蔗糖轉化酶可以催化蔗糖水解成葡萄糖與果糖，提供細胞能量使用或形成訊號分子。在細胞質中的蔗糖轉化酶在非酸性環境有較高的活性，因此又被稱為中鹼性蔗糖轉化酶。在阿拉伯芥中共有九種中鹼性蔗糖轉化酶，其中CINV1與CINV2為最主要的中鹼性蔗糖轉化酶。過去研究發現這類蔗糖轉化酶參與植物根部生長發育、滲透壓逆境調節、開花及細胞壁生合成等途徑。除此之外，CINV1胺基酸序列第547的位置被認為是能與14-3-3蛋白交互作用的磷酸化位點。本研究發現在鹽分處理後，磷酸化蛋白質體分析發現CINV1具第547胺基酸位點的磷酸化胜肽片段比例上升。本研究欲探究CINV1是否影響植物生長發育或逆境反應。在鹽分處理後，CINV1的突變株其主根具有較大的生長偏角。而於鹽分處理情況下外加醣類 (葡萄糖、果糖、海藻糖)，其主根生長偏角則與野生型類似。另外，觀察互補株在鹽分處理後的主根生長偏角，發現性狀可以被回復，與野生型類似。此外，將Ser547點突變之後，蛋白質在細胞中的表現位置會改變。最後，利用雙分子螢光互補系統驗證CINV1可以和translationally controlled tumor protein (TCTP)有交互作用，且外加target of rapamycin (TOR)途徑的抑制劑之後，CINV1突變株主根長的縮短幅度較野生型來的小，且側根密度增加，暗示CINV1可能參與TOR訊號傳遞路徑。 | zh_TW |
| dc.description.abstract | Sucrose is the major form of nutrient in plant transport. Invertase catalyzes the hydrolysis of sucrose to glucose and fructose, which provides cell energy or as a signaling molecule. Invertases in the cytoplasm are highly active in a non-acidic environment and are therefore also referred to as alkaline/neutral invertases (A/N-Invs). There are nine alkaline/neutral invertases in Arabidopsis, of which CINV1 and CINV2 are the major ones. Previous studies have found that CINV1 is involved in root growth and development, regulation of osmotic stress, flowering and cell wall synthesis. In addition, Ser547 of CINV1 is considered to be a phosphorylation site that can interact with 14-3-3 proteins. This study found that after salt treatment, the phosphorylated peptides containing Ser547 increased abundance. The present study aims to study whether Ser547 phosphorylation is important for plant growth and stress responses. It was found that under salt stress condition, CINV1 mutants have larger primary root deviation angle from vertical orientation than wild type. The growth angle can be recover by addition of glucose, fructose, or trehalose, but not by sucrose. In complementation lines of CINV1, deviation angle of primary root was recovered. This study also found point mutation of Ser547 led to subcellular localization of CINV1 change. In addition, the bimolecular fluorescence complementation assay showed that CINV1 can interact with translationally controlled tumor protein (TCTP). After adding target of rapamycin (TOR) inhibitor, primary root length of CINV1 mutant was less affect than wild type, and the lateral root density was increased, which suggests that CINV1 is involved in TOR signaling pathway. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T06:00:00Z (GMT). No. of bitstreams: 1 ntu-108-R04b42013-1.pdf: 5434602 bytes, checksum: 7faab05794ddcaf5420982550df6abad (MD5) Previous issue date: 2019 | en |
| dc.description.tableofcontents | 誌謝 I
摘要 II Abstract III Abbreviation IV Contents V List of Table IX List of Figures X Appendixes XII 1. Introduction 1 1.1 Biotic and abiotic stresses 1 1.1.1 Osmotic stress 1 1.1.2 Salt stress and the impact to plants 1 1.2 Invertases in plants 2 1.2.1 Alkaline/neutral invertases (A/N-Invs) in Arabidopsis 2 1.2.2 Cytosolic invertase1 (CINV1) 2 1.3 Root growth and development 4 1.3.1 Architecture of primary roots 4 1.3.2 Lateral root development 5 1.3.3 Directional growth of roots 5 1.4 Target of rapamycin (TOR) pathway 5 1.4.1 Inhibitors of TOR pathway 6 1.5 14-3-3 proteins 6 1.6 Project goals 7 2. Materials and Methods 8 2.1 Plant materials 8 2.2 Growth conditions 8 2.3 Generation of constructs 8 2.4 Arabidopsis transformation by floral dipping 9 2.5 Isolation of T-DNA insertional mutants 10 2.6 RNA extraction and gene expression analysis 11 2.6.1 Semi-quantitative and quantitative real-time PCR analysis 11 2.7 Detection of plant protein expression level 12 2.7.1 Plant protein extraction and concentration measurement 12 2.7.2 Western blot 12 2.8 Primary root length and lateral root density measurement 14 2.9 Root directional growth measurement 14 2.10 Root meristem measurement 14 2.11 Isolation of Arabidopsis leaf protoplasts 15 2.12 Bimolecular Fluorescence Complementation (BiFC) 15 2.12.1 Plasmid construction for BiFC analysis 15 2.12.2 Transformation of plasmids for BiFC analysis in Arabidopsis protoplasts 16 2.12.3 BiFC analysis using Nicotiana benthamiana leaf system 16 2.13 Carbohydrate content 17 2.14 Phosphoproteome analysis 18 2.14.1 Sample preparation 18 2.14.2 Protein extraction and in-solution trypsin digestion 18 2.14.3 Desalting and phosphopeptides enrichment 19 2.14.4 Mass spectrum analysis and protein identification 20 2.15 Statistical analysis 20 3. Results 21 3.1 Transcript and protein level of CINV1 affected by salt treatment 21 3.2 Isolation of cinv1 mutants 22 3.3 Primary root length was less affected in cinv1 mutants under salt stress condition 23 3.4 Directional growth was enhanced in cinv1 mutants when treated with sodium chloride 24 3.5 Sucrose and trehalose content in cinv1 mutants was higher than wild type under salt stress condition 24 3.6 Application of sugar or salt induces root deviation from vertical orientation 25 3.7 Gravity response of mutants was higher than wild type under salt stress condition 25 3.8 Importance of Ser547 phosphorylation site of CINV1 in lateral root density under 3% mannitol-treated condition 26 3.9 Importance of Ser547 phosphorylation site of CINV1 in directional growth under salt-treated condition 27 3.10 Mutation of Ser547 phosphorylation site appears to change the subcellular localization of CINV1 27 3.11 Point-mutation variants of CINV1 can interact with 14-3-3ω 28 3.12 CINV1 can interact with TCTP 28 3.13 CINV1 might be involved in TOR pathway 28 4. Discussion 29 4.1 Phosphorylation of CINV1 29 4.2 Root growth affected by CINV1 30 References 33 Table 39 Figures 40 Appendixes 65 | |
| dc.language.iso | en | |
| dc.subject | 中鹼性蔗糖轉化? | zh_TW |
| dc.subject | 鹽分逆境 | zh_TW |
| dc.subject | 蛋白質磷酸化 | zh_TW |
| dc.subject | 根 | zh_TW |
| dc.subject | 甘露醇 | zh_TW |
| dc.subject | alkaline/neutral invertase | en |
| dc.subject | phosphorylation | en |
| dc.subject | root | en |
| dc.subject | mannitol | en |
| dc.subject | salt stress | en |
| dc.title | 阿拉伯芥蔗糖轉化酶CINV1磷酸化與根部生長發育的功能性研究 | zh_TW |
| dc.title | Functional Study of Arabidopsis Invertase CINV1 Phosphorylation in Root Growth and Development | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 107-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 王淑珍,洪傳揚,張世宗,鄭秋萍 | |
| dc.subject.keyword | 中鹼性蔗糖轉化?,鹽分逆境,蛋白質磷酸化,根,甘露醇, | zh_TW |
| dc.subject.keyword | alkaline/neutral invertase,salt stress,phosphorylation,root,mannitol, | en |
| dc.relation.page | 76 | |
| dc.identifier.doi | 10.6342/NTU201900522 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2019-02-13 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 植物科學研究所 | zh_TW |
| 顯示於系所單位： | 植物科學研究所 | |
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