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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張英峯(Ing-Feng Chang) | |
| dc.contributor.author | Chun-Yi Hsu | en |
| dc.contributor.author | 許淳奕 | zh_TW |
| dc.date.accessioned | 2022-11-24T09:26:52Z | - |
| dc.date.available | 2022-11-24T09:26:52Z | - |
| dc.date.copyright | 2021-11-04 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-10-21 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81759 | - |
| dc.description.abstract | WRKY轉錄因子在逆境下扮演重要的角色，也被報導與多種蛋白會有交互作用。WRKY為植物特有之轉錄因子，可透過與w-box結合活化或是抑制以調節下游基因表現，並對植物生理功能上做出相對應的反應。透過雙分子螢光互補實驗發現14-3-3w及HDA6對於WRKY25有交互作用，其功能還不明確。另外，為了瞭解HDA6上的磷酸化位點為何，我們也透過激酶活性分析發現其片段中三到四個位點對於磷酸化似乎有著其重要性。為了研究WRKY25與HDA6交互作用之功能為何，我們將hda6突變株 (hda6-6) 與WRKY25大量表現株進行雜交。在鹽處理下，WRKY25大量表現株與35S:WRKY25/hda6-6與野生型相比有較高的生存率，hda6-6則是對野生型有相對低的生存率，表示35S:WRKY25/hda6-6抗鹽的表現型貢獻自WRKY25。另外我們也針對開花時間進行測試。過去文獻中發現hda6-6有晚開花的現象，而在實驗中發現WRKY25大量表現株會有早開花的現象，而35S:WRKY25/hda6-6則是介於兩者之間，表示大量表現WRKY25似乎能部分互補hda6-6對於開花的功能，使其開花時間提前發生。另外，也針對WRKY25潛在的磷酸化位點進行突變，我們發現到似乎WRKY25 第345號蘇胺酸為14-3-3w與WRKY25結合位，並且第345號蘇胺酸點突變株在鹽逆境下的生存率是有所差異的。綜合上述結果，阿拉伯芥中WRKY25可能透過與HDA6及14-3-3w之交互作用，並對於鹽分逆境與開花上有所調控。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-24T09:26:52Z (GMT). No. of bitstreams: 1 U0001-2010202111165000.pdf: 3771632 bytes, checksum: f217e5f116c2bda74ef5b182b7d673e2 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | "誌謝 I 摘要 II Abstract III Abbreviations V List of tables XI List of figures XII Supplementary data XIII Appendix XIV 1. Introduction 1 1.1 Environmental stresses 1 1.1.1 Salt stress responses in plants 1 1.1.2 Sensing and signal transduction in salt stress 2 1.2 WRKY transcription factors 3 1.2.1 WRKY25 4 1.2.2 WRKY TFs in plant development 5 1.2.3 WRKY TFs in abiotic stress responses 6 1.2.4 Interacting partners of WRKYs 6 1.2.5 Phosphorylation of WRKY TFs 7 1.3 14-3-3 proteins…. 8 1.4 Histone deacetylases 9 1.4.1 HISTONE DEACETYLASE 6 10 1.5 Project goals 11 2. Materials and Methods 12 2.1 Plant materials 12 2.2 Growth conditions 12 2.3 Generation of constructs for WRKY25 overexpression lines 13 2.4 Arabidopsis transformation by floral dipping 14 2.5 Isolation of T-DNA insertional mutants and splice site mutant 15 2.6 RNA extraction and gene expression analysis 16 2.7 Semi-quantitative and quantitative real-time PCR analysis 16 2.8 Measurement of survival rates and seed germination rates 17 2.9 Phenotyping of flowering days and rosette leaf number 17 2.10 Isolation of Arabidopsis leaf protoplasts 18 2.11 Bimolecular Fluorescence Complementation (BiFC) assay 19 2.11.1 BiFC analysis using Arabidopsis protoplast 19 2.11.2 BiFC analysis in N. benthamiana 20 2.12 Purification of recombinant protein 20 2.13 Protein quantification by Bradford assay 21 2.14 In vitro kinase assay 22 2.15 Total protein extraction 23 2.16 Immunoprecipitation and in gel digestion 23 2.17 In-solution trypsin digestion 24 2.18 Statistical analysis 25 3. Results 26 3.1 Isolation of wrky25 mutant and WRKY25 overexpression lines 26 3.2 Interactions between WRKY25, 14-3-3 and HDA6 26 3.3 Thr424/Thr425/Ser427/Ser429 of HDA6 might be phosphorylated by CDPK16 in vitro 27 3.4 Isolation of splicing mutant hda6-6 and cross lines of hda6 and WRKY25 overexpression line 29 3.5 35S:WRKY25/hda6-6 exhibits higher salt tolerant 30 3.6 35S:WRKY25/hda6-6 showed early flowering phenotype 31 3.7 Mutation of Thr345 site of WRKY25 to Ala affects Arabidopsis salt tolerance 32 4. Discussion 33 4.1 WRKY25 and HDA6 are involved in flowering 33 4.2 35S:WRKY25/hda6-6 exhibits higher tolerant than hda6-6 under salt stress condition 35 4.3 Function of the phosphorylation sites of HDA6 37 5. References 39 Table 45 Figures 54 Supplementary data 69 Appendixes 78" | |
| dc.language.iso | en | |
| dc.subject | 14-3-3 | zh_TW |
| dc.subject | WRKY25 | zh_TW |
| dc.subject | HDA6 | zh_TW |
| dc.subject | 鹽逆境 | zh_TW |
| dc.subject | 磷酸化 | zh_TW |
| dc.subject | 生存率 | zh_TW |
| dc.subject | 開花 | zh_TW |
| dc.subject | WRKY25 | en |
| dc.subject | survival rate | en |
| dc.subject | phosphorylation | en |
| dc.subject | salt stress | en |
| dc.subject | HDA6 | en |
| dc.subject | 14-3-3 | en |
| dc.subject | flowering | en |
| dc.title | WRKY25與HDA6交互作用對於鹽分逆境反應及開花的影響 | zh_TW |
| dc.title | Effects of interaction of WRKY25 and HDA6 in salt stress response and flowering in Arabidopsis | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 吳克強(Hsin-Tsai Liu),謝旭亮(Chih-Yang Tseng),鄭貽生,謝明勳 | |
| dc.subject.keyword | WRKY25,HDA6,鹽逆境,磷酸化,生存率,開花,14-3-3, | zh_TW |
| dc.subject.keyword | WRKY25,HDA6,salt stress,phosphorylation,survival rate,flowering,14-3-3, | en |
| dc.relation.page | 84 | |
| dc.identifier.doi | 10.6342/NTU202103918 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2021-10-23 | |
| 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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