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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林讚標(Tsan-Piao Lin) | |
| dc.contributor.author | Chia-Hao Liu | en |
| dc.contributor.author | 劉家豪 | zh_TW |
| dc.date.accessioned | 2021-06-16T08:23:38Z | - |
| dc.date.available | 2014-03-08 | |
| dc.date.copyright | 2014-03-08 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-01-24 | |
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Herbivory rapidly activates MAPK signaling in attacked and unattacked leaf regions but not between leaves of Nicotiana attenuata. Plant Cell 19, 1096-1122. Zhai, Q., Yan, L., Tan, D., Chen, R., Sun, J., Gao, L., Dong, MQ., Wang, Y., Li, C. (2013). Phosphorylation-Coupled Proteolysis of the Transcription Factor MYC2 Is Important for Jasmonate-Signaled Plant Immunity. PLoS Genet 9, 1003422 Zarei, A., Korbes, A. P., Younessi, P., Montiel, G., Champion, A., Memelink, J. (2011). Two GCC boxes and AP2/ERF-domain transcription factor ORA59 in jasmonate/ethylene-mediated activation of the PDF1.2 promoter in Arabidopsis. Plant Mol Biol 75, 321-331. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/58643 | - |
| dc.description.abstract | 目前為止的研究報導已經知道有很多轉錄因子參與茉莉酸(JA)訊息傳遞的調控，像是MYC2和一些ERFs。ORA47是一個ERF轉錄因子，並且在wounding和JA誘導的情況下會大量表現，我們研究室的研究也已經發現ORA47是很多JA生合成基因的上游調控者。我的研究主題是找到可能的ORA47上游調控者。在核甘酸序列分析發現ORA47 promoter序列上含有一些保守性序列，像是MYC2結合的保守性序列(CACGTG, or G-box)和推測的WRKY26和WRKY38結合序列(TTGATC, or W-like box)，因此我們想知道它們在wounding訊息傳遞中是否會直接調控ORA47表現。ORA47 pro:GUS 感染(inoculated) 實驗觀察到myc2突變株的GUS活性比野生型植物來的低，代表MYC2對wounding誘導的訊息傳遞是必要的。ORA47和一些JA生合成基因在35S::MYC2-GFP轉植株中的表現量都被誘導表現，代表MYC2是ORA47的上游調控者。染色體免疫共沉澱(ChIP)分析實驗增加MYC2可以和ORA47 promoter上兩個位置相近的G-box結合，接著電泳遷移改變分析法(EMSA)也看到MYC2只能結合到G-box1，並且酵母單雜交分析(yeast one-hybrid)結果也看到MYC2會和G-box1結合而不是G-box2；關於WRKY轉錄因子的部份，WRKY26和WRKY38在電泳遷移改變分析法實驗中都可以結合到W-like-box2 (WL-box2)、W-like-box3、W-like-box4位置上，並且也有微弱的結合到W-like-box1的能力。在酵母單雜交分析分析則看到WRKY26和WRKY38可以和WL-box1還有WL-box4結合。染色體免疫共沉澱分析則看到WRKY26和WRKY38只能和ORA47 promoter的WL-box1結合。本篇研究的結論是：MYC2，WRKY26和WRKY38在JA訊息傳遞路徑上是ORA47的直接上游調控者。 | zh_TW |
| dc.description.abstract | Jasmonates (JAs) are plant signaling molecules that play important roles in defense of biotic stress. Many transcription factors, such as MYC2 and some ERFs are reported to be involved in the regulation of the signaling pathways mediated by JA. ORA47 is an ERF transcription factor and its expression is highly induced by wounding and JA. ORA47 is also known from our lab to upregulate many JA biosynthetic genes. My research aim is to study the potential upstream components of ORA47. Nucleotide sequence analysis revealed that the promoter of the ORA47 gene contained conserved sequence motifs, which are analogous to the MYC2 binding consensus sequences (CACGTG, or G-box) and putative WRKY26 and WRKY38 binding sequences (TTGATC, or W-like box). We therefore asked whether they could directly regulate ORA47 expression in wounding signaling. The GUS activity of ORA47 pro:GUS inoculated plants was lower in myc2 mutant line than in WT indicating wounding induced ORA47 expression is MYC2-dependent. ORA47 and several genes involved in JA biosynthesis were induced in the 35S::MYC2-GFP mutants compared with WT plant indicating MYC2 is an upstream regulator of JA biosynthesis. Chromatin immunoprecipitation (ChIP) assay showed MYC2 could interact with two closely linked G-box elements in the ORA47 promoter. Furthermore, electrophoretic mobility shift assay (EMSA) analysis showed MYC2 could only bind to G-box1 and yeast one-hybrid assay also exihibted MYC2 could interact with the G-BOX1 fragment but not G-BOX2. With respect to WRKY transcription factors, both WRKY26 and WRKY38 could bind to W-like-box2 (WL-box2), W-like-box3, W-like-box4 and much less the W-like- box1 cis-elements in the EMSA experiment. Moreover, yeast one hybrid assay showed WRKY26 and WRKY38 could interact with the WL-box1 and WL-box4 DNA fragments. ChIP assay revealed WRKY26 and WRKY38 could interact only with the WL-box1 element in the ORA47 promoter. In conclusion, MYC2, WRKY26, and WRKY38 directly regulate ORA47 in the JA signaling pathway. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T08:23:38Z (GMT). No. of bitstreams: 1 ntu-103-R00b42022-1.pdf: 2819684 bytes, checksum: 34e7ded05f8b4d4f7380e5b8cb3e9323 (MD5) Previous issue date: 2014 | en |
| dc.description.tableofcontents | 摘要 8
Abstract 9 縮寫對照表 11 第一章 序論 13 1.1 Wounding stress 13 1.2 Jasmonates…………………………………………………………13 1.3 JA biosynthesis………. 14 1.4 JA signal pathway………… 14 1.5 MAPK signal pathway………………. 15 1.6 ORA47…………… 16 1.7 MYC2….. 16 1.8 WRKY……………. 17 1.9 研究目標……………………………………………………………18 第二章 材料與方法 19 2.1 植物材料、生長條件 19 2.2 基因序列分析 19 2.3 DNA萃取及Polymerase Chain Reaction (PCR)聚合酶連鎖反應 19 2.4 RNA萃取及cDNA合成之reverse transcriptase PCR (RT-PCR) 21 2.5 即時定量聚合酶連鎖反應 (real-time PCR) 22 2.6 轉形構築及轉基因植物的建立 24 2.7 Yeast one hybrid質體建構 25 2.8 染色質免疫沈澱法(chromatin immunoprecipitation) 27 2.9 GST-WRKY26和GST-MYC2蛋白純化 28 2.10 膠體電泳位移分析(electrophoretic mobility shift assay, EMSA) 28 2.11 染酵母單雜交分析(yeast one-hybrid) 31 第三章 結果 33 3.1 利用inoculation表現GUS基因的方法進行初步分析 33 3.2 利用real-time PCR的方式分析35S::MYC2-GFP轉殖珠的基因表現 33 3.3 利用ChIP分析MYC2在植物體內和ORA47 promoter不同位置的結合能力 34 3.4 利用EMSA分析MYC2對不同ORA47 promoter DNA片段的結合能力 34 3.5 利用yeast one hybrid分析MYC2是否會和ORA47 promoter結合 34 3.6 利用EMSA分析WRKY26對不同ORA47 promoter DNA片段的結合能力 35 3.7 利用yeast one hybrid分析WRKY26和WRKY38是否會和ORA47 promoter結合 35 3.8 利用ChIP分析WRKY26和WRKY38在植物體內和ORA47 promoter不同位置的結合能力 36 第四章 討論 37 參考文獻 41 圖表 45 附錄 61 | |
| dc.language.iso | zh-TW | |
| dc.title | "在傷害誘導的JA訊息傳遞過程中，WRKY26, WRKY38 和 MYC2是調控ORA47基因表現的上游轉錄因子" | zh_TW |
| dc.title | WRKY26, WRKY38 and MYC2 are direct upstream transcription factors regulating ORA47 gene expression in wound-induced JA signaling | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 鄭石通(Shih-Tong Jeng),吳克強(Ke-qiang Wu),吳素幸(Shu-Hsing Wu),謝明勳(Ming-Hsiun Hsieh) | |
| dc.subject.keyword | 茉莉酸,MYC2,WRKY26,WRKY38,ORA47,傷害逆境,阿拉伯芥, | zh_TW |
| dc.subject.keyword | JA,MYC2,WRKY26,WRKY38,ORA47,wounding,arabidopsis, | en |
| dc.relation.page | 70 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2014-01-24 | |
| 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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