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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 金洛仁(Laurent Zimmerli) | |
| dc.contributor.author | Jingsong Zhang | en |
| dc.contributor.author | 張靜松 | zh_TW |
| dc.date.accessioned | 2021-06-16T02:29:13Z | - |
| dc.date.available | 2020-08-01 | |
| dc.date.copyright | 2015-08-28 | |
| dc.date.issued | 2015 | |
| dc.date.submitted | 2015-07-31 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/53765 | - |
| dc.description.abstract | Ethylene response factors (ERFs)的轉錄因子已經被證實參與在阿拉伯芥(Arabidopsis thaliana)的生長發育以及防禦反應中。在本研究中，我們發現了一個未被報道的ERF轉錄因子19。它是一個可能結合在啟動子區的GCC序列上的轉錄因子，並且能夠負調控阿拉伯芥對於灰黴菌的防禦反應。大量表現ERF19會降低茉莉酸防禦反應的標誌基因PDF1.2a和PR3的基因表現，并使植株對於灰黴菌更加感病。儘管在原生質體轉化實驗(protoplast transactivation assay)中發現, ERF19是轉錄上的激活因子，但是在生物體內的bimolecular fluorescence complementation實驗和co-immunoprecipitation實驗結果表明ERF19和NINJA抑制子以及组蛋白脱乙酰酶HDA6，HDA19複合體交互作用。此外原生質體轉化實驗表明ERF19的轉錄活性被NINJA顯著性的抑制了。因此，我們猜測ERF19通過與NINJA抑制子複合體交互作用負調控茉莉酸反應。我們的實驗結果闡述了ERF19介導的一個新的轉錄抑制機制。 | zh_TW |
| dc.description.abstract | Arabidopsis ethylene response factors (ERFs) are known to be involved in various developmental and defense processes. Here, we report novel mechanisms of ERF19, a potential GCC box binding transcription factor, to negatively regulate defense response against the fungal pathogen Botrytis cinerea. Over-expression of ERF19 leads to increased Arabidopsis susceptibility to B. cinerea via repressing the induction of jasmonic acid (JA) defense marker genes PDF1.2a and PR3. Although ERF19 acted as activator of transcription in protoplast transactivation assay (PTA), in vivo bimolecular fluorescence complementation (BiFC) and co-immunoprecipitation (Co-IP) assays revealed that ERF19 associates with the transcriptional repressor NINJA and histone deacetylases HDA6 and HDA19. Moreover, ERF19 activity were significantly repressed by NINJA in PTA assay. We proposed that ERF19 recruits the NINJA repressor complex to negatively regulate JA-mediated defense responses. Our results provide new insights into the mechanisms of ERF19-mediated transcriptional repression. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T02:29:13Z (GMT). No. of bitstreams: 1 ntu-104-R02b42031-1.pdf: 3950482 bytes, checksum: 3d461e65939f3e165084424b60f61b02 (MD5) Previous issue date: 2015 | en |
| dc.description.tableofcontents | 摘要 i Abstract ii Contents iii Introduction 5 Material and Methods 10 Biological Materials and Growth Conditions. 10 Disease assays. 10 Transgenic plants. 11 qRT-PCR. 11 Protoplast transactivation assay. 12 Co-immunoprecipitation Assay in Arabidopsis Protoplast. 12 Bimolecular Fluorescence Complementation Assay in Arabidopsis Protoplast. 13 Chemical Treatments. 14 Protein Gel Blotting Analysis 14 Root length measurements. 15 Statistical analysis. 15 Results 16 Generation of ERF19 inducible over-expression and RNAi lines. 16 ERF19 acts as an activator of transcription. 16 ERF19 associates with HDA6, HDA19, and NINJA in vivo. 17 NINJA, but not HDA6 and HDA19, represses the activation of ERF19. 18 A role for NINJA in disease resistance. 19 Chitin treatment causes ERF19 degradation and this degradation is dependent on 26S proteasome. 19 Characterization of erf20 mutant and ERF20 over-expression lines. 20 Discussion 22 Conclusion and future perspectives 24 Figures 26 Figure 1: Introduction of ERF19 with GFP into the pMDC7 binary vector. 26 Figure 2: Introduction of Micro ERF19 into the pMDC7 binary vector 27 Figure 3: Introduction of NINJA, HDA6, and HDA19 into the p35SN vector and ERF19 into the p35S-GAL4DB vector. 28 Figure 4: ERF19 acts as an activator of transcription. 29 Figure 5: ERF19 associates with HDA6, HDA19, and NINJA when analyzed by the BiFC assay in Arabidopsis protoplasts. 31 Figure 6: ERF19 associates with HDA6, HDA19, and NINJA when tested by Co-IP in Arabidopsis protoplasts. 32 Figure 7: NINJA, but not HDA6 and HDA19, represses the activation of ERF19. 33 Figure 8: Disease resistance of ninja RNAi and NINJA OE lines to B. cinerea. 35 Figure 9: Degradation of ERF19 by chitin treatment. 36 Figure 10: ERF19 is subjected to proteasome-mediated degradation. 37 Figure 11: The identification of erf20 mutant and overexpression line. 38 Figure 12: ERF20 associates with NINJA and NHDA19 when analyzed by BiFC in Arabidopsis protoplasts. 40 Figure 13: Reduced root growth in untreated erf20 mutant line 41 Figure 14: Alignment of ERF19 and ERF20 protein sequences 42 Figure 15:Proposed model 43 Tables 44 Table 1: Primers for inducible erf19 RNAi line construct 44 Table 2: Primers for BiFC assay construct 44 Table 3: Primers for CoIP assay construct 45 Table 4: Primers for PTA assay construct 46 Table 5: Primers for qRT-PCR 46 References 47 | |
| dc.language.iso | en | |
| dc.subject | NINJA抑制子 | zh_TW |
| dc.subject | ERF轉錄因子19 | zh_TW |
| dc.subject | 灰黴菌 | zh_TW |
| dc.subject | 茉莉酸反應 | zh_TW |
| dc.subject | ERF19 | en |
| dc.subject | NINJA | en |
| dc.subject | JA-mediated responses | en |
| dc.subject | Botrytis cinerea | en |
| dc.title | "乙烯反應因子19與NINJA抑制子複合體交互作用,負調控阿拉伯芥對於灰黴菌的防禦反應" | zh_TW |
| dc.title | The ethylene response factor ERF19 associates with the NINJA repressor complex and negatively regulates Arabidopsis defense responses against Botrytis cinerea | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 103-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 吳克強,林?標 | |
| dc.subject.keyword | ERF轉錄因子19,灰黴菌,茉莉酸反應,NINJA抑制子, | zh_TW |
| dc.subject.keyword | ERF19,Botrytis cinerea,JA-mediated responses,NINJA, | en |
| dc.relation.page | 53 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2015-07-31 | |
| 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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