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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 劉瑞芬 | |
| dc.contributor.author | Ｗei-Che Shih | en |
| dc.contributor.author | 施維哲 | zh_TW |
| dc.date.accessioned | 2021-06-17T02:28:49Z | - |
| dc.date.available | 2022-08-28 | |
| dc.date.copyright | 2017-08-28 | |
| dc.date.issued | 2017 | |
| dc.date.submitted | 2017-08-18 | |
| dc.identifier.citation | 黃俊慈，2014 以蛋白體學分析菸草中參與Elicitin作用之SlSOBIR1交互作用蛋白，臺灣大學植物病理與微生物學研究所學位論文
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/68645 | - |
| dc.description.abstract | 植物藉由位於細胞膜之pattern-recognition receptor (PRR) 辨認環境中微生物的保守性構造(microbe-associated molecular pattern, MAMP)，而引發Pattern-triggered immunity (PTI)。 近年研究發現PRR的作用尚須其他PRR交互作用蛋白的參與，例如細胞膜蛋白BRI1-associated receptor kinase (BAK1)與Suppressor of BIR1-1 (SOBIR1)等。疫病菌(Phytophtora parasitica)是植物重要病原菌，目前已有elicitin ParA1 (P. parasitica elicitin)等多種疫病菌的MAMP被發現。Elicitin能在多種茄科植物引起植物組織壞疽（necrosis），BAK1與 SOBIR1皆被發現在elicitin引發植物細胞死亡過程扮演不可或缺的角色。SlSOBIR1原本位於細胞膜，以ParA1處理植物後，會被胞吞進入胞內體（endosome）。為瞭解相關作用機制，黃(2014)以SlSOBIR1進入胞內體的時機點進行共免疫沉澱及質譜分析，發現菸草NbRLP1可能為SlSOBIR1的交互作用蛋白。本研究進一步分析其特性，發現接種疫病菌後24小時，疫病菌在菸草引發的水浸狀壞疽病斑開始擴大，NbRLP1的基因表現量也大幅提升。以agrobacterium-mediated transient expression過量表現 NbRLP1能加劇ParA1 在菸草葉片引發組織壞疽，而靜默NbRLP1的基因表現則會抑制疫病菌於菸草的病程發展。以共軛焦顯微鏡觀察NbRLP1-GFP在菸草葉片表皮細胞的分佈情形，發現NbRLP1-GFP位於內質網膜，且因應ParA1的處理會集中分佈於內質網與細胞膜接觸點 (endoplasmic reticulum-plasma membrane contact site, EPCS) ，進一步將SOBIR1-mCherry與NbRLP1-GFP 共同表現於菸草葉片發現，因應ParA1處理，SOBIR1-mCherry所標記的胞內體會緊靠著NbRLP1-GFP所標示的內質網與EPCS構造分佈。這些結果顯示NbRLP1位於內質網膜，但可能在EPCS與SOBIR1產生交互作用；此外，NbRLP1也在植物與疫病菌的交互作用及植物對ParA1的反應扮演重要角色，但相關詳細機制尚待深入探討。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2021-06-17T02:28:49Z (GMT). No. of bitstreams: 1 ntu-106-R03633004-1.pdf: 43980672 bytes, checksum: 2db85211ff0c081d64875969dc0ee281 (MD5) Previous issue date: 2017 | en |
| dc.description.tableofcontents | 摘要 I
Abstract III 前言 1 一、植物防禦反應 1 1.3先天性免疫反應 (innate immunity) 1 1.2、Pattern-recognition receptor (PRR) 2 1.3、PRR 交互作用蛋白 3 二、疫病菌 5 2.1、 疫病菌生活史和發病生態 5 2.2、 疫病菌侵染模式 6 2.3、 疫病菌的 Microbe-associated molecular pattern (MAMP) 6 2.4、Elicitin 7 三、內質網與植物抗病 8 3.1、內質網壓力 (ER stress) 8 3.2 、內質網壓力與病原引起之計畫性細胞死亡(Program cell death, PCD) 9 3.3、細胞膜與內質網連結 (ER-PM contact site, EPCS) 10 四、研究動機及策略 11 貳、材料與方法 12 ㄧ、供試植物與生長條件 12 二、疫病菌侵染實驗 12 2.1 菌株來源與保存 12 2.2 疫病菌產孢與游走子收集 12 2.3 疫病菌的接種 13 2.4 菸草病害指數評分 13 三、植物基因的表現量分析 14 3.1 收集植物組織樣本 14 3.2 RNA萃取 與製備 cDNA 14 3.3 即時定量聚合酶鏈鎖反應 (quantitative RT-PCR) 14 四、運用Tobacco rattle virus (TRV)靜默 NbRLP1表現 15 4.1 靜默載體構築 15 4.2 農桿菌轉殖 16 4.3 農桿菌感染法(Agroinfection) 16 五、分析 NbRLP1 於細胞中的分佈 17 5.1 NbRLP1 C端融合 GFP的重組載體構築 17 5.2 ER-PM contact site (EPCS) 標定基因 SYT1與 VAP27 之融合蛋白載體構築 17 5.3 在菸草葉片以農桿菌短暫大量表現重組蛋白 18 5.4 疫病菌 ParA1 的處理 18 5.5 以雷射掃描式共軛焦螢光顯微鏡 (Confocal microscope) 進行觀察 19 六、 親緣性分析 19 6.1 NbRLP1的胺基酸序列分析 19 參、結果 20 ㄧ、NbRLP1序列分析 20 二、NbRLP1之親緣關係分析 21 三、菸草接種疫病菌後 NbRLP1 表現量大幅提升 21 四、增強NbRLP1之基因表現加劇ParA1造成之細胞壞疽 22 五、靜默NbRLP1之基因表現能抑制疫病菌侵染菸草之病徵 23 六、NbRLP1-GFP 分佈於內質網且其分佈情形因應 ParA1 處理產生變化 24 七、NbRLP1-GFP 所標示之內質網串珠狀構造為內質網—細胞膜接觸位置（endoplasmic reticulum-plasma membrane contact site, EPCS） 25 八、SlSOBIR1-mCherry 聚集於 NbRLP1-GFP 分佈之內質網構造和 EPCS 旁 26 肆、討論 27 一、NbRLP1參與菸草-疫病菌的交互作用 27 二、NbRLP1參與 eilcitin 於茄科作物引發之 細胞壞疽 27 三、NbRLP1 為內質網膜蛋白且其分佈因應 ParA1 處理發生改變。 29 四、植物內質網於病原菌侵染時之動態變化 30 五、EPCS 參與於植物接受胞外訊息後的向內傳遞中 31 六、HR 於菸草與疫病菌之互動中所扮演的角色。 31 七、結語 32 伍、參考文獻 33 陸、附表 41 七、附圖 43 捌、補充資料 55 | |
| dc.language.iso | zh-TW | |
| dc.subject | 圓葉菸草 | zh_TW |
| dc.subject | 疫病菌 | zh_TW |
| dc.subject | Elicitin | zh_TW |
| dc.subject | PTI | zh_TW |
| dc.subject | NbRLP1 | zh_TW |
| dc.subject | SlSOBIR1 | zh_TW |
| dc.subject | Phytophthora parasitica | en |
| dc.subject | SlSOBIR | en |
| dc.subject | Nicotiana benthamiana | en |
| dc.subject | elicitin | en |
| dc.subject | PTI | en |
| dc.subject | NbRLP1 | en |
| dc.title | 探討基因NbRLP1 在圓葉菸草與疫病菌交互作用的角色 | zh_TW |
| dc.title | The Role of NbRLP1 in the Response of Nicotiana benthamiana to the Elicitin ParA1 | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 105-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 王昭雯,鄭秋萍,趙光裕 | |
| dc.subject.keyword | 圓葉菸草,疫病菌,Elicitin,PTI,NbRLP1,SlSOBIR1, | zh_TW |
| dc.subject.keyword | Nicotiana benthamiana,Phytophthora parasitica,PTI,NbRLP1,SlSOBIR,elicitin, | en |
| dc.relation.page | 58 | |
| dc.identifier.doi | 10.6342/NTU201703717 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2017-08-18 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 植物病理與微生物學研究所 | zh_TW |
| 顯示於系所單位： | 植物病理與微生物學系 | |
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