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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊淑怡(Shu-Yi Yang) | |
| dc.contributor.author | Yi-Hsien Wei | en |
| dc.contributor.author | 魏禕嫺 | zh_TW |
| dc.date.accessioned | 2022-11-24T09:25:55Z | - |
| dc.date.available | 2022-11-24T09:25:55Z | - |
| dc.date.copyright | 2022-02-21 | |
| dc.date.issued | 2022 | |
| dc.date.submitted | 2022-01-27 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81700 | - |
| dc.description.abstract | 植物的側根經常會在與叢枝菌根菌( arbuscular mycorrhizal fungi)共生的時候增加。在我們過去的研究中已經發現，菌根菌共生會使番茄中的C-terminally encoded peptides (CEPs)，SlCEP2，表現量下降。外源施用於根部的生長素會減輕由脯胺酸上具有羥基化修飾的CEP2 (CEP2Hyp)胜肽處理所造成的側根形成缺陷，顯示CEP2會通過影響根部生長素相關的路徑來負調控側根形成。SlCEPR1 (CEP receptor 1)可能為CEP2的受體。同樣的，菌根菌共生可能會減輕CEP2胜肽的負面影響來增加側根形成。本研究的主旨在於更進一步探討CEP2、CEPR1以及另一個也會被菌根菌共生負調節的CEP3，在菌根菌共生促進的側根生長中的作用。本研究發現CEP2的表現會可能受到由菌根菌所提供的氮源影響。成熟CEP2胜肽表現在細胞核、細胞質與細胞膜，並且CEP2胜肽可能並不會被運輸到植物的地上部。菌根菌共生與CEP2Hyp胜肽在側根原基形成上具有相反的功能。在根部施用IAA可以減輕CEP2Hyp所造成的側根形成缺陷。相反的，在地上部施用NAA卻不能恢復因CEP2Hyp處理而下降的植物側根密度，表示CEP2Hyp胜肽主要影響是在側根發育扮演重要角色且是來自根部的生長素。與病毒誘導的CEP2基因靜默 (CEP2-VIGS) 植物相反，CEPR1-VIGS植物的側根密度仍會被菌根菌共生誘導增加，顯示CEP2的基因表現量對菌根菌共生促進的側根形成可能更為重要。大量表現CEP3會抑制菌根菌增加側根形成的能力，雖然其確切機制仍然未知。在我們的系統中，CEP2、CEPR1和CEP3對於菌根菌的生長發育非常微弱或無明顯影響。這些結果有助於了解菌根菌共生是如何通過調節CEP胜肽來增進番茄側根形成。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-24T09:25:55Z (GMT). No. of bitstreams: 1 U0001-2701202214472000.pdf: 4163230 bytes, checksum: 7a4d0b81748e3eea5faf147dcf0e32ce (MD5) Previous issue date: 2022 | en |
| dc.description.tableofcontents | "摘要 iii Abstract iv List of abbreviation vi Table of Contents ix List of Tables xii List of Figures xiii List of Supplemental Figures xiv List of Appendixes xv Chapter 1 Introduction 1 1.1 The importance of arbuscular mycorrhizal (AM) symbiosis 1 1.2 Lateral root growth is enhanced during AM symbiosis 2 1.3 The potential role of auxin in AM symbiosis-stimulated lateral root growth 6 1.4 C-terminally encoded peptide (CEP) involves symbiosis regulation and lateral root growth 9 1.5 AM symbiosis may enhance tomato lateral root formation through modulating the expression of SlCEP2 peptide 14 1.6 The aim of the research 14 Chapter 2 Materials and methods 16 2.1 Plant materials and growth conditions 16 2.2 Synthetic CEP2Hyp peptide, IAA and shoot NAA treatment 16 2.3 Measurement of lateral root density and length 17 2.4 Observation of unemerged lateral root primordia 18 2.5 Imaging of auxin distribution 18 2.6 Construct generation 19 2.7 Amplification of CEP2 promoter by FPNI-PCR 20 2.8 Promoter sequence analysis for potential cis-elements 21 2.9 Subcellular localization of CEP2 21 2.10 Root staining and quantification of AM fungal colonization level 22 2.11 RNA extraction, cDNA synthesis, and qRT-PCR analysis 22 2.12 Western blot 23 2.13 Virus-mediated transient expression 24 2.14 Accession numbers 25 Chapter 3 Results 26 3.1 High nitrate induced the expression of CEP2 in tomato roots 26 3.2 The cis-elements in CEP2 promoter region 28 3.3 Subcellular localization of CEP2 30 3.4 Lateral root formation altered by AM symbiosis and CEP2Hyp 31 3.5 IAA application on roots restored the LR phenotype affected by CEP2Hyp treatment 32 3.6 CEP2Hyp-caused LR defect was not mitigated by application of NAA on shoot apex 34 3.7 The role of CEPR1 in AM-enhanced LR formation 35 3.8 Transcription factor binding sites on CEPR1 promoter 37 3.9 CEP3 may have a potential contribution to CEP2-leaded LR defects 39 Chapter 4 Discussion 41 4.1 AM fungi-provided nitrate might influence CEP2 expression 41 4.2 The potential regulator of CEP2 remains to be clarified 42 4.3 Subcellular localization of CEP2 is detected on nuclear, cytoplasm and plasma membrane 43 4.4 CEP2 shows an opposite effect on LR formation compared to AM symbiosis 45 4.5 CEP2-mediated LR defect in response to exogenous auxin 46 4.6 The role of CEPR1 in AM symbiosis-enhanced LR formation 47 4.7 The regulation of CEPR1 is still unclear 49 4.8 More evidence is required to understand the function of CEP3 50 4.9 Overall conclusion of this study 52 Tables 53 Figures 62 Supplemental Figures 78 Appendixes 83 References 85 Table 1. Recipe of 1/2 Hoagland solution. 53 Table 2. Primers used in vector construction 55 Table 3. Gene specific primers (GSPs) used in 3’RACE. 56 Table 4. Fusion primers (FPs), FP-specific primers (FSPs) and gene specific primers (GSPs) used in FPNI-PCR 57 Table 5. Primers used in qRT-PCR. 58 Table 6. Information of CEP genes in tomato 60 Figure 1. Fungal colonization and gene expression in response to different nitrate concentrations and R. irregularis. 62 Figure 2. CEP2 promoter cloning results using fusion primer and nested integrated PCR (FPNI-PCR). 63 Figure 3. Prediction of putative cis-elements in the promoter analysis of SlCEP2.. 64 Figure 4. Investigating EGFP-tagged CEP2 fusion protein in CEP2-EGFP overexpressed hairy root tomatoes by western blot. 65 Figure 5. Subcellular localization of CEP2 in tomato root cells 66 Figure 6. Undetectable CEP2-EGFP signal in tomato leaves 67 Figure 7. LR and unemerged LRP phenotype in roots under AM symbiosis. 68 Figure 8. LR and unemerged LRP phenotype in roots in response to CEP2Hyp treatment. 69 Figure 9. Tomato LR growth in response to CEP2Hyp peptide and IAA treatment. 70 Figure 10. Effects of CEP2Hyp treatment and shoot NAA application on tomato LR development. 71 Figure 11. Expression of CEPR1 in response to different nitrate conditions and AM colonization. 72 Figure 12. LR development, colonization level and gene expression of CEPR1 knock-down plants in response to AM symbiosis. 73 Figure 13. Prediction of putative cis-elements in the promoter analysis of CEPR1. 74 Figure 14. Gene structure of CEP3, and its expression under inoculation of AM fungi. 75 Figure 15. Effects of CEP3 overexpression on root phenotype and gene expression. 76 Figure 16. Overall model of this study. 77 Supplemental Figure 1. CEP2 gene expression in response to R. irregularis (Ri) and different phosphate conditions in tomato roots (Provide by Yu-Heng Hsieh). 78 Supplemental Figure 2. Subcellular localization of CEP2 in N. benthamiana cells (Provide by Yu-Heng Hsieh). 79 Supplemental Figure 3. Tomato LR development under CEP2Hyp peptide, IBA and NAA treatment (Provide by Yu-Heng Hsieh). 80 Supplemental Figure 4. Effects of CEP2 expression level on LR phenotype, fungal colonization level and gene expression (Provide by Yu-Heng Hsieh). 81 Supplemental Figure 5. Root phenotype and CEPR1 expression of CEPR1-VIGS tomato root in response to CEP2Hyp treatment (Provide by Yu-Heng Hsieh). 82 Appendix 1. Schematic summary of IAA biosynthesis pathway 83 Appendix 2. Recent research about CEPs in Arabidopsis and M.truncatula 84" | |
| dc.language.iso | en | |
| dc.subject | 生長素 | zh_TW |
| dc.subject | 叢枝菌根菌 | zh_TW |
| dc.subject | 胜肽荷爾蒙 | zh_TW |
| dc.subject | 胜肽受體 | zh_TW |
| dc.subject | 側根形成 | zh_TW |
| dc.subject | arbuscular mycorrhizal symbiosis | en |
| dc.subject | C-terminally encoded peptide (CEP) | en |
| dc.subject | auxin | en |
| dc.subject | lateral root formation | en |
| dc.subject | peptide hormone | en |
| dc.title | 探討番茄的CEP2/3胜肽參與叢枝菌根菌促進側根形成之作用 | zh_TW |
| dc.title | Deciphering the role of tomato CEP2/3 peptides involved in AM-enhanced lateral root formation | en |
| dc.date.schoolyear | 110-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 謝旭亮(Chien-Hua Huang),鄭秋萍(Nien-Tzu Chang),陳賢明(Ssu-Yuan Chen),林雅芬(Chin-Hao Chang) | |
| dc.subject.keyword | 叢枝菌根菌,胜肽荷爾蒙,胜肽受體,側根形成,生長素, | zh_TW |
| dc.subject.keyword | arbuscular mycorrhizal symbiosis,C-terminally encoded peptide (CEP),lateral root formation,peptide hormone,auxin, | en |
| dc.relation.page | 100 | |
| dc.identifier.doi | 10.6342/NTU202200230 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2022-01-29 | |
| 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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