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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 金洛仁(Laurent Zimmerli) | |
| dc.contributor.author | Beier Jiang | en |
| dc.contributor.author | 蔣貝爾 | zh_TW |
| dc.date.accessioned | 2021-07-11T14:44:50Z | - |
| dc.date.available | 2021-08-24 | |
| dc.date.copyright | 2016-08-24 | |
| dc.date.issued | 2016 | |
| dc.date.submitted | 2016-08-01 | |
| dc.identifier.citation | Acosta, I.F., Gasperini, D., Chetelat, A., Stolz, S., Santuari, L., and Farmer, E.E. (2013). Role of NINJA in root jasmonate signaling. Proceedings of the National Academy of Sciences of the United States of America 110, 15473-15478.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/78177 | - |
| dc.description.abstract | 植物在受到病原菌攻擊時會啟動免疫反應從而引起一系列免疫基因的表達。這個過程需要非常精細的轉錄水平上的重新編碼。舉例來說，乙烯調控因子在植物調節免疫反應中有著重要的作用。在我們實驗室之前的研究中發現，阿拉伯芥過表達ERF19基因時表現出比野生型更加容易被灰霉病菌感染的性狀。這種性狀是通過抑制免疫基因PDF1.2a以及PR3的表達所導致的。在我的研究中，構建了ERF19的功能缺失植株，發現它們表現出比野生型更加抗灰霉病菌感染的性狀。通過原生質體瞬時表達實驗發現ERF19是轉錄激活子而ERF19過表達植株中的免疫基因被抑制。於是我通過酵母雙雜交，蛋白質免疫共沉澱以及雙分子螢光光互補實驗發現ERF19蛋白可以和NINJA抑制複合體有交互作用。更有趣的是，在ninja突變體中過表達ERF19蛋白會嚴重改變阿拉伯芥的生長表型以及抗病表型。所有這些新的發現對於研究ERF轉錄因子參與植物免疫反應有嶄新的意義。 | zh_TW |
| dc.description.abstract | When attacked by pathogens, plants activate the immune responses to regulate the inducible expression of a large set of defense genes. This process requires effective transcriptional reprogramming. For example, ethylene responsive factor (ERF) transcription factors (TFs) have been shown to play critical roles in regulating immune responses in plants. In the Zimmerli laboratory, overexpression of ERF19 was shown to increase Arabidopsis susceptibility to Botrytis cinerea (B. cinerea) via repression of the induction of the jasmonic acid (JA) defense marker genes PDF1.2a and PR3. In this study, I generated pERF19:ERF19-EAR transgenic lines and these loss-of-function plants exhibited increased resistance to B. cinerea, when compared to wild type (WT) plants. ERF19 acted as a transcriptional activator, however defense genes were repressed in ERF19 overexpression lines. Using the yeast two hybrid, Co-IP and BiFC assays, ERF19 was shown to associate with the NINJA repressor complex. Moreover, overexpression of ERF19 in Arabidopsis ninja mutant strongly changes the developmental pattern and defense phenotype of the plant. Notably, a strong increased susceptibility to B. cinerea was observed. All these findings provide new insight on how ERF TFs regulate plant immunity. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-11T14:44:50Z (GMT). No. of bitstreams: 1 ntu-105-R03b42032-1.pdf: 2139015 bytes, checksum: c8ca13789c8e19e4fce71d95eb076b4d (MD5) Previous issue date: 2016 | en |
| dc.description.tableofcontents | Contents
誌謝 i 摘要 ii Abstract iii Contents iv Introduction 1 Arabidopsis defenses against B. cinerea 1 Pathogen-associated molecular patterns-triggered immunity and effector-triggered immunity 1 Chitin-triggered immunity 2 ERF transcription factors 3 Roles of ERF proteins in biotic stress 4 The role of NINJA in JA signaling 4 EAR motif-mediated transcriptional repression in plants 5 Mechanism of action of the co-repressor TPL 6 HDACs participate in the defense response 6 Materials and Methods 8 Plant materials and growth conditions 8 Transgenic plants 8 Pathogen infection assay 9 β-estradiol induction before pathogen infection 9 PAMP treatment 9 Genomic DNA extraction 10 RNA extraction and gene expression analysis 10 Protoplast transactivation assay 11 Transient expression in Nicotiana Benthamiana 11 Subcellular localization in Nicotiana benthamiana leaves 12 Yeast two-hybrid assays 12 Bimolecular Fluorescence Complementation assays 12 Co-immunoprecipitation assay in Arabidopsis protoplast 13 SDS-PAGE and Immunoblotting 13 Accession numbers 14 Results 15 Loss of ERF19 function is linked with enhanced resistance to B. cinerea 15 pERF19:ERF19-SRDX transgenic lines display enhanced expression of defense gene 15 Generation of Arabidopsis ERF19 artificial microRNA transgenic lines 16 Subcellular localization of ERF19 in Nicotiana benthamiana 17 ERF19 directly interacts with NINJA in vitro 17 ERF19 associates with the NINJA repressor complex in vivo 18 ERF19 associates with HDA6 and HDA19 in vivo 18 Arabidopsis ninja mutant exhibit enhanced resistance while NINJA overexpression lines are more susceptible to B. cinerea 19 Overexpress ERF19 in Arabidopsis ninja mutant showed altered developmental phenotype 20 Lines overexpressing ERF19 in the ninja mutant exhibits strong increased susceptibility to B. cinerea 20 Discussion 21 Future Perspectives 24 Figures 25 Figure 1. Disease responses of pERF19:ERF19-SRDX transgenic plants to B. cinerea infection. 25 Figure 2. Response of ERF19 to amiRNA. 27 Figure 3. Subcellular localization of ERF19. 28 Figure 4. Direct interaction of ERF19 with NINJA but not with TPL, HDA6 or HDA19 by Y2H 29 Figure 5. ERF19 associates with TPL in vivo. 30 Figure 6. Association of ERF19 with NINJA, HDA6 and HDA19 but not with TPL by BiFC 31 Figure 7. Disease responses of ninja mutant and NINJA overexpression plants to B. cinerea infection. 32 Figure 8. Development and defense phenotype of ninja mutant overexpressing ERF19. 33 Figure 9. Model for ERF19-mediated defense repression in Arabidopsis. 34 Supplementary Figures 35 Figure S1. Constitutive and inducible overexpression lines of ERF19 were more susceptible to B. cinerea infection than WT. 35 Figure S2. ERF19 associates with NINJA, HDA6 and HDA19 by Co-IP in Arabidopsis protoplast. 36 Figure S3. NINJA repress the activation of ERF19. 37 Tables 39 Table 1. Mutant alleles used in this study 39 Table 2. Primers used for ninja mutant check 39 Table 3. Primers used for ERF19 amiRNA plasmid construction 39 Table 4. Primers used for plasmid construction 40 Table 5. Primers used for qRT-PCR 40 | |
| dc.language.iso | en | |
| dc.title | ERF19和NINJA抑制複合體參與阿拉伯芥抗灰霉病的功能性分析 | zh_TW |
| dc.title | Functional analysis of ERF19 and NINJA repressor complex involved in plant defense against Botrytis cinerea | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 104-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 吳克強(Keqiang Wu),鄭貽生(Yi-Sheng Cheng),王雅筠(Ya-Yun Wang) | |
| dc.subject.keyword | 阿拉伯芥,灰黴菌,乙烯反應因子,NINJA抑制子,自然免疫, | zh_TW |
| dc.subject.keyword | Arabidopsis thaliana,Botrytis cinerea,ethylene responsive factor,NINJA repressor complex,innate immunity, | en |
| dc.relation.page | 48 | |
| dc.identifier.doi | 10.6342/NTU201601705 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2016-08-02 | |
| 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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