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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳昭瑩(Chao-Ying Chen) | |
| dc.contributor.author | Po-Yang Huang | en |
| dc.contributor.author | 黃博洋 | zh_TW |
| dc.date.accessioned | 2022-11-25T05:33:53Z | - |
| dc.date.available | 2023-08-18 | |
| dc.date.copyright | 2021-11-11 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-08-20 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/82002 | - |
| dc.description.abstract | "植物會因應病原菌侵擾而活化先天性免疫，受到侵擾的組織會累積水楊酸(salicylic acid, SA)並長距離傳訊，啟動系統性誘導抗病反應(systemic acquired resistance, SAR)。LsGRP1為介導葵百合抗百合灰黴病的關鍵防禦蛋白，內源性表現及外源性施用LsGRP1皆可有效抑制處理葉及系統葉百合灰黴病的病徵，指出LsGRP1除了誘導原位抗性也具有誘導植物系統性抗病的能力，然其機制仍有待釐清。前人研究發現阿拉伯芥葉片表現LsGRP1可提升病原菌入侵後所誘導的防禦反應，而本研究進一步觀察外源性大腸桿菌生產之融合蛋白SUMO-LsGRP1△SS的作用，發現其可促使阿拉伯芥葉片提升由flg22激活的免疫反應；在系統葉上可減少Pseudomonas syringae pv. tomato (Pst) DC3000的增殖菌量及增加flg22誘發的癒傷葡聚醣沉積。經試驗得知SUMO-LsGRP1△SS之部分區段刪除突變的誘導系統性抗病強度皆較SUMO-LsGRP1△SS者為低。藉由阿拉伯芥誘導系統性抗病的水楊酸依賴型傳訊路徑(SA-dependent signaling pathway)之sid2 (SA induction deficient 2)、bsmt1 (benzoic acid/SA carboxyl methyltransferase 1)突變株及2-哌啶甲酸依賴型傳訊路徑(pipecolic acid-dependent signaling pathway) 之ald1 (AGD2-like defense response protein 1)、sard4 (SAR-deficient 4)突變株分析比較SUMO-LsGRP1△SS誘導系統葉抗Pst DC3000的強度，得知這兩型傳訊路徑皆參與LsGRP1誘導系統性抗病之發生，且SUMO-LsGRP1△SS可提升水楊酸依賴型傳訊路徑之SID2及BSMT1基因及2-哌啶甲酸依賴型傳訊路徑之ALD1及SARD4基因的表現量，確認LsGRP1在阿拉伯芥處理葉激活這兩型系統性抗病傳訊路徑。綜合言之，LsGRP1可以誘導處理葉及系統葉產生抗病反應，且經由水楊酸及2-哌啶甲酸依賴型傳訊路徑啟動系統性抗病的表現，此等研究為LsGRP1之植物保護應用發展建立了重要的學理基礎。" | zh_TW |
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| dc.description.tableofcontents | "中文摘要 I 英文摘要 II 壹、前言 1 貳、前人研究 3 一、植物先天性免疫與系統性誘導抗病(Plant innate immunity and systemic acquired resistance) 3 植物系統性誘導抗病 3 二、植物激活物 (Plant activator) 4 三、百合防禦蛋白LsGRP1 5 四、植物第二型富含甘胺酸蛋白 (Plant glycine-rich proteins, GRPs) 5 五、植物源蛋白質或胜肽誘導植物抗性 6 六、LsGRP1與系統性誘導抗病 7 七、植物系統性抗病傳訊路徑(SAR signaling pathways) 7 水楊酸依賴型傳訊路徑(SA-dependent signaling pathway) 7 2-哌啶甲酸依賴型傳訊路徑(Pip-dependent signaling pathway) 8 參、材料與方法 10 一、植物栽培 10 二、病原菌培養與接種 10 1. 番茄細菌性斑點病接種 10 2. 草莓葉枯病菌接種 10 三、農桿菌浸潤法 (Agroinfiltration) 11 四、細菌病原分子模式flg22之製備 11 五、SUMO-LsGRP1△SS與其不同區段刪除突變之製備 12 1. 大腸桿菌表現LsGRP1衍生性蛋白之菌株 12 2. 融合蛋白表現及純化 12 六、十二烷基硫酸鈉聚丙烯醯胺膠體電泳偵測蛋白大小 13 1. 十二烷基硫酸鈉聚丙烯醯胺膠體電泳 13 2. 西方墨點法 14 七、以農桿菌原位葉短暫表現LsGRP1於阿拉伯芥誘導系統性抗病 14 八、注射SUMO-LsGRP1△SS於阿拉伯芥葉片誘導系統性抗病 15 九、阿拉伯芥葉片癒傷葡聚醣之植物組織染色法 15 1. 處理葉 15 2. 系統葉 16 十、SUMO-LsGRP1△SS不同區段刪除突變對誘導阿拉伯芥系統性抗病之影響 16 十一、以突變株探討LsGRP1誘導系統性抗病與傳訊路徑關聯性 16 十二、偵測阿拉伯芥基因之相對表現量 17 1. 阿拉伯芥PTI相關基因及SAR傳訊路徑相關基因之表現量偵測 17 2. 阿拉伯芥RNA萃取 17 3. 反轉錄-即時聚合酶連鎖反應 17 十三、評估澆灌含SUMO-LsGRP1ΔSS之大腸桿菌溶解產物施用溶液於田間防治草莓葉枯病之效果 18 1. 製備含SUMO-LsGRP1ΔSS之大腸桿菌溶解產物施用溶液 18 2. 澆灌SUMO-LsGRP1ΔSS之大腸桿菌溶解產物施用溶液防治草莓葉枯病之田間試驗 18 肆、結 果 20 一、外源處理SUMO-LsGRP1△SS可以促進阿拉伯芥的免疫活化 20 二、以農桿菌浸潤法內源性短暫表現LsGRP1可以系統性強化阿拉伯芥抗病性與flg22誘導的癒傷葡聚醣沉積 20 三、浸潤完整的SUMO-LsGRP1△SS才能有效誘發植物系統性抗病 21 四、水楊酸依賴型傳訊路徑基因突變會破壞浸潤SUMO-LsGRP1△SS誘導之系統性抗病 22 五、2-哌啶甲酸依賴型傳訊路徑基因突變會破壞浸潤SUMO-LsGRP1△SS誘導之系統性抗病 23 六、浸潤SUMO-LsGRP1△SS於阿拉伯芥可以活化水楊酸依賴型傳訊路徑SID2及BSMT1基因表現 23 七、浸潤SUMO-LsGRP1△SS於阿拉伯芥可以活化2-哌啶甲酸依賴型傳訊路徑ALD1及SARD4基因表現 24 八、澆灌SUMO-LsGRP1ΔSS的大腸桿菌粗萃液防治田間草莓病害之效果 25 伍、討 論 26 陸、參考文獻 31 柒、圖表集 42 表一、反轉錄-即時聚合酶連鎖反應引子對 43 圖一、以SUMO-LsGRP1△SS處理阿拉伯芥葉片明顯增加由flg22誘導的癒傷葡聚醣沉積 44 圖二、以SUMO-LsGRP1△SS處理阿拉伯芥葉片明顯增加flg22誘導之PTI標誌基因FRK1表現 46 圖三、於阿拉伯芥葉片內源性表現LsGRP1可減少系統葉Pst DC3000的增殖菌量 47 圖四、於阿拉伯芥葉片內源性表現LsGRP1可增加系統葉由flg22誘導之癒傷葡聚醣沉積 48 圖五、以西方墨點分析大腸桿菌系統表現之SUMO-LsGRP1△SS及其不同區段刪除突變 50 圖六、以SUMO-LsGRP1△SS處理阿拉伯芥葉片可減少系統葉之Pst DC3000增殖菌量 51 圖七、以SUMO-LsGRP1△SS處理阿拉伯芥葉片可增加系統葉上flg22誘導之癒傷葡聚醣沉積 52 圖八、比較SUMO-LsGRP1△SS及其不同區段刪除突變於阿拉伯芥誘導系統性抗病的程度 54 圖九、在阿拉伯芥水楊酸依賴型傳訊路徑之突變株上，以SUMO-LsGRP1△SS處理無法減少系統葉Pst DC3000的增殖菌量 55 圖十、在阿拉伯芥2-哌啶甲酸依賴型傳訊路徑之突變株上，以SUMO-LsGRP1△SS處理無法減少系統葉Pst DC3000的增殖菌量 57 圖十一、以SUMO-LsGRP1△SS處理可誘導阿拉伯芥水楊酸依賴型傳訊路徑之SID2與BSMT1基因表現 59 圖十二、以SUMO-LsGRP1△SS處理可誘導阿拉伯芥2-哌啶甲酸依賴型傳訊路徑之ALD1與SARD4基因表現 61 圖十三、SUMO-LsGRP1ΔSS蛋白粗萃液抑制草莓葉枯病之田間試驗 64" | |
| dc.language.iso | zh-TW | |
| dc.subject | 2-哌啶甲酸依賴型傳訊路徑 | zh_TW |
| dc.subject | LsGRP1 | zh_TW |
| dc.subject | 阿拉伯芥 | zh_TW |
| dc.subject | 系統性抗病 | zh_TW |
| dc.subject | 水楊酸依賴型傳訊路徑 | zh_TW |
| dc.subject | pipecolic acid-dependent signaling pathway | en |
| dc.subject | systemic acquired resistance | en |
| dc.subject | salicylic acid-dependent signaling pathway | en |
| dc.subject | LsGRP1 | en |
| dc.subject | Arabidopsis thaliana | en |
| dc.title | LsGRP1誘導阿拉伯芥系統性抗病之傳訊路徑探討 | zh_TW |
| dc.title | Investigation of the signaling pathways for LsGRP1-induced systemic disease resistance in Arabidopsis thaliana | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 沈偉強(Hsin-Tsai Liu),鄭秋萍(Chih-Yang Tseng),林乃君 | |
| dc.subject.keyword | LsGRP1,阿拉伯芥,系統性抗病,水楊酸依賴型傳訊路徑,2-哌啶甲酸依賴型傳訊路徑, | zh_TW |
| dc.subject.keyword | LsGRP1,Arabidopsis thaliana,systemic acquired resistance,salicylic acid-dependent signaling pathway,pipecolic acid-dependent signaling pathway, | en |
| dc.relation.page | 65 | |
| dc.identifier.doi | 10.6342/NTU202102427 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-08-23 | |
| 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 |
| dc.date.embargo-lift | 2023-08-18 | - |
| 顯示於系所單位： | 植物病理與微生物學系 | |
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