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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 靳宗洛 | zh_TW |
| dc.contributor.advisor | Tsung-Luo Jinn | en |
| dc.contributor.author | 魏品萱 | zh_TW |
| dc.contributor.author | Pin-Xuan Wei | en |
| dc.date.accessioned | 2025-02-13T16:22:08Z | - |
| dc.date.available | 2025-02-14 | - |
| dc.date.copyright | 2025-02-13 | - |
| dc.date.issued | 2025 | - |
| dc.date.submitted | 2025-02-10 | - |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/96415 | - |
| dc.description.abstract | 氣候變化導致自然災害頻發，如高溫、洪水、土壤鹽鹼化和長期乾旱，給全球各地帶來了巨大挑戰。非生物逆境，包括乾旱、高鹽、極端溫度及水淹等因素，已成為全球農業生產的主要問題，影響植物生長和作物產量，並對糧食安全構成威脅。植物在非生物逆境的應對過程中，需要啟動調控機制來維持細胞穩定性。熱休克轉錄因子 (HSFs)，在植物對多種非生物逆境反應中扮演重要角色。除了在高溫逆境中表現突出外，HSFs也廣泛參與植物應對乾旱、鹽鹼及低溫等多重逆境條件的反應，負責啟動下游逆境防禦基因的表達，促進植物的適應能力。為了更加了解HSFA7a/b在非生物逆境下扮演的角色，我們利用CRISPR-Cas9技術建立HSFA7a 與 HSFA7b的雙突變株。結果表明，HSFA7a 和 HSFA7b 在不同的非生物逆境條件下均被顯著誘導表達，顯示出它們可能參與植物多種非生物逆境反應。透過耐熱性、離層酸處理後的發芽狀況以及鹽處理後子葉綠化狀況試驗，證實 HSFA7a 負向調控逆境反應，而HSFA7b 扮演正向調控的角色。值得注意的是，兩者的單突變體呈現出相反的表現型，反映了這兩者在逆境反應中的功能差異。為了進一步解析HSFA7a/b的功能，我們透過雙分子螢光互補實驗以及冷光素酶互補成像分析確認了HSFA7a/b與 HSFA2 之間的蛋白質-蛋白質相互作用，並透過雙螢光素酶報導基因分析證實了它們在APX2基因表達調控中的協同作用。這些結果顯示，HSFA7a/b與HSFA2共同形成一個協同調控網路，幫助植物在非生物逆境中保持穩定，增強其適應能力。最後，RNA-seq分析中差異表達基因 (DEGs) 的重疊顯示，HSFA7a/b可能與植物清除活性氧反應有著密切相關。總結來說，我們的研究探討HSFA7a/b在植物應對多種非生物逆境中的調控機制，並識別出潛在的下游基因。 | zh_TW |
| dc.description.abstract | Climate change has resulted in more natural disasters, including higher temperatures, floods, soil salinization, and droughts, creating significant challenges for global communities. Abiotic stresses such as drought, high salinity, and extreme temperatures have a severe impact on agriculture and food security. Heat shock transcription factors (HSFs) are crucial in helping plants respond to these stresses by regulating defense gene expression. In our study, we used CRISPR-Cas9 to create double mutants of the HSFA7a and HSFA7b genes to explore their roles in abiotic stress responses. Our findings revealed that both genes are activated by heat, abscisic acid (ABA), salt stress, and cold stress. Specifically, HSFA7a negatively regulates responses to abiotic stress, while HSFA7b acts as a positive regulator. Protein interactions between HSFA7a/b and HSFA2 were confirmed using BiFC and LCI assays, and dual-luciferase reporter assays showed their involvement in regulating the APX2 gene. Additionally, RNA-seq analysis indicated their role in reactive oxygen species (ROS) scavenging. Overall, our research emphasizes the roles of HSFA7a/b in plant responses to abiotic stresses and identifies potential downstream genes involved. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2025-02-13T16:22:08Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2025-02-13T16:22:08Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 致謝 i
摘要 ii ABSTRACT iii ABBREVIATIONS ix INTRODUCTION 1 Heat Stress/Shock (HS) 1 Heat shock factors (HSFs) 2 Heat shock proteins (HSPs) 5 Heat shock-related genes 5 Salt stress 6 Abscisic acid (ABA) response 7 OBJECTIVES 9 MATERIALS AND METHODS 10 Plant Materials and Growth Conditions 10 Generation of the HSFA7a/b-Overexpression (OE) and HSFA7b-Complementation (COM) Transgenic Plants 11 RNA Preparation, cDNA Synthesis and Real-Time Quantitative PCR 11 Protein Extraction 13 HSFA7a and HSFA7b Promoter::galactosidase (GUS) Expression in Transgenic Arabidopsis Plants 13 Thermotolerance Tests 14 Seed Germination, Post-germination Seedling Growth Assay 15 Dual-Luciferase Reporter (DLR) Assay 15 Bimolecular Fluorescence Complementation (BiFC) Assay and Firefly Luciferase Complementation Imaging (LCI) Assay 16 Western Blot Analysis 17 RNA-seq Analysis 18 Statistical Analysis 19 Primer and Accession Number 20 RESULTS 21 HSFA7a and HSFA7b were expressed in various tissues in Arabidopsis 21 HSFA7a and HSFA7b were induced by multiple abiotic stresses 21 HSFA7a and HSFA7b interacted with HSFA2 23 HSFA7a and HSFA7b existed self-interaction and heterologous interaction 24 Characterization of HSFA7a and HSFA7b T-DNA insertion mutant 24 Generation of HSFA7a and HSFA7b double mutant by CRISPR/Cas9 25 Generation of HSFA7a and HSFA7b over-expression transgenic plants 25 Short-term and long-term acquired thermotolerance analysis for mutants 26 The cotyledon greening rate of HSFA7a and HSFA7b mutants in response to salt stress 27 The seed germination rate of HSFA7a and HSFA7b mutants in response to ABA stress 28 Expression of HS-related genes in responses to HS in HSFA7a- and HSFA7b-mutant plants 28 HSFA7a and HSFA7b cooperated with HSFA2 to activate the transcription of APX2 29 Transcriptome analysis in response to HS in the hsfa7a mutant 29 Transcriptome analysis in response to HS in the hsfa7b mutant 30 Transcriptome analysis in response to HS in the Dou-IS mutant 31 DISCUSSION 32 HSFA7a/b are rapidly induced by HS 32 HSFA7a/b may be involved in various stress crosstalk 33 The phenotypical analysis for mutants in response to salt and ABA stress 36 HSFA7a/b may cooperate with HSFA2 to regulate the mechanism of HSR 39 RNA-seq analysis 40 Conclusions and Prospects 41 TABLE 43 FIGURES 45 SUPPLEMENTAL FIGURES 84 REFERENCES 93 | - |
| dc.language.iso | en | - |
| dc.subject | 核糖核酸定序 | zh_TW |
| dc.subject | 活性氧 | zh_TW |
| dc.subject | 非生物逆境 | zh_TW |
| dc.subject | 阿拉伯芥 | zh_TW |
| dc.subject | 熱休克轉錄因子 | zh_TW |
| dc.subject | Reactive oxygen species | en |
| dc.subject | Heat shock transcription factor | en |
| dc.subject | Arabidopsis | en |
| dc.subject | Abiotic stress | en |
| dc.subject | RNA sequencing | en |
| dc.title | 阿拉伯芥熱休克因子HsfA7a/b在非生物逆境反應之功能性研究 | zh_TW |
| dc.title | Functional study on Arabidopsis heat shock factors HsfA7a/b in abiotic stress response | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 113-1 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 葉靖輝;楊健志;鄭秋萍;鄭石通 | zh_TW |
| dc.contributor.oralexamcommittee | Ching-Hui Yeh;Chien-Chih Yang;Chiu-Ping Cheng;Shih-Tong Jeng | en |
| dc.subject.keyword | 熱休克轉錄因子,阿拉伯芥,非生物逆境,核糖核酸定序,活性氧, | zh_TW |
| dc.subject.keyword | Heat shock transcription factor,Arabidopsis,Abiotic stress,RNA sequencing,Reactive oxygen species, | en |
| dc.relation.page | 105 | - |
| dc.identifier.doi | 10.6342/NTU202500536 | - |
| dc.rights.note | 同意授權(限校園內公開) | - |
| dc.date.accepted | 2025-02-10 | - |
| dc.contributor.author-college | 生命科學院 | - |
| dc.contributor.author-dept | 植物科學研究所 | - |
| dc.date.embargo-lift | 2030-02-08 | - |
| 顯示於系所單位： | 植物科學研究所 | |
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