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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 劉瑞芬(Ruey-Fen Liou) | |
| dc.contributor.author | Pei-Yi Lai | en |
| dc.contributor.author | 賴沛宜 | zh_TW |
| dc.date.accessioned | 2022-11-25T05:36:23Z | - |
| dc.date.available | 2027-02-09 | |
| dc.date.copyright | 2022-02-18 | |
| dc.date.issued | 2022 | |
| dc.date.submitted | 2022-02-09 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/82125 | - |
| dc.description.abstract | "OPEL為疫病菌 (Phytophthora parasitica) 的外泌蛋白，序列預測顯示其具有signal peptide、thaumatin-like domain、glycine-rich domain及GH16 domain，GH16 domain包含β-1,3-glucanase活性區的保守性序列。以OPEL重組蛋白處理菸草 (Nicotiana tabacum cv. Samsun-NN) 葉片能引發細胞死亡、癒傷葡聚醣沉積、活性氧分子累積及誘導水楊酸和PTI相關防禦基因表現。對處理OPEL重組蛋白之菸草葉片所進行的轉錄體分析發現多個編碼receptor-like proteins (RLPs) 或receptor-like kinases (RLKs) 的差異表現基因 (FC ≥ 2, p-value < 0.01)，為瞭解其在OPEL誘發植物免疫的重要性，本研究挑選具研究潛力者，包括被高度誘導表現的NtLYM2、涉及多種醣分子辨識的adaptor RLKs 基因NtCERK1-like以及參與蛋白質辨識的NtSOBIR1和NtBAK1等；RT-qPCR分析顯示這些基因的表現都會因應OPEL之處理顯著提升。進一步以cucumber mosaic virus (CMV) 於N. tabacum cv. Samsun-NN誘導基因靜默，再分析其對OPEL誘發癒傷葡聚醣沉積和細胞離子滲漏的影響，發現靜默NtLYM2顯著減少OPEL介導癒傷葡聚醣累積的數量，靜默NtBAK1降低OPEL引發的細胞離子滲漏量，而靜默NtCERK-like或NtSOBIR1對二項觀測指標皆無顯著影響。此外，疫病菌感染誘導NtLYM2大量表現，而且暫表現NtLYM2顯著提升疫病菌在菸草的感染面積。這些研究顯示NtLYM2和NtBAK1在OPEL引發的植物免疫反應扮演重要角色，至於NtLYM2如何參與菸草和疫病菌的交互作用則待更多研究探討。" | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-25T05:36:23Z (GMT). No. of bitstreams: 1 U0001-0902202223063700.pdf: 3147907 bytes, checksum: 553b188b9e41e37c0bdcd24499ce8791 (MD5) Previous issue date: 2022 | en |
| dc.description.tableofcontents | 口試委員會審定書 i 誌 謝 ii 摘 要 iv Abstract v 壹、前言 1 1. 植物防禦反應 1 2. MAMPs 2 3. DAMPs 3 4. PRR及PRR複合體 (PRR complex) 4 4.1 LRR-RLPs/RLKs 5 4.2 LysM-RLPs/RLKs 6 4.3 Lectin-like和EGF-like RLPs/RLKs 6 5. 病原菌細胞壁分解酵素在植物防禦反應之角色 7 6. β-葡聚醣 (β-glucan) 引發植物基礎免疫反應 9 7. 疫病菌 10 8. 疫病菌外泌蛋白OPEL引發植物基礎免疫反應 12 9. 研究動機 13 貳、材料與方法 14 1. 供試植物 14 2. 親緣關係分析 14 3. 表現與純化OPEL重組蛋白 14 3.1 OPEL重組蛋白表現轉形株來源與保存 14 3.2 重組蛋白表現與純化 14 3.3 凝膠過濾層析 (Gel filtration chromatography) 15 4. 植物RNA純化與cDNA製備 16 4.1 萃取植物RNA 16 4.2 去除殘留於RNA分離液的DNA 16 4.3 cDNA製備 16 5. 即時定量聚合酶連鎖反應 (RT-qPCR) 17 6. 基因靜默表現載體構築 17 7. 暫表現 (transient expression) 載體構築 18 8. 大腸桿菌轉形 19 9. 農桿菌轉形 19 10. 農桿菌浸潤 20 10.1 以農桿菌感染法 (agroinfection) 進行病毒誘導系統性基因靜默 20 10.2 以農桿菌浸潤法 (agroinfiltration) 暫表現基因 20 11. 西方墨點法 (Western blot) 21 11.1 植物組織蛋白質萃取與聚丙烯醯胺膠體電泳 (SDS-PAGE) 21 11.2 蛋白質轉漬 21 11.3 抗體雜合與訊號偵測 21 12. 癒傷葡聚醣沉積 (callose deposition) 分析 22 13. 細胞離子滲漏測定 22 14. 疫病菌接種 23 14.1 N. tabacum cv. Samsun-NN疫病菌P. parasitica全株接種 23 14.2 N. benthamiana疫病菌P. parasitica離葉接種 23 參、結果 24 1. NtLYM2、NtCERK1、NtSOBIR1和NtBAK1同源性基因親緣關係分析 24 2. OPEL重組蛋白純化與其引發植物防禦反應的活性 26 3. NtLYM2、NtCERK1-like、NtSOBIR1和NtBAK1因應OPEL處理之表現情形 26 4. 測試CMV誘導N. tabacum基因靜默系統於的效率 27 5. 基因靜默NtLYM2、NtCERK1-like、NtSOBIR1或NtBAK1對OPEL引發植物基礎免疫反應的影響 28 6. 疫病菌P. parasitica感染誘導NtLYM2的基因表現 29 7. 暫表現NtLYM2提升疫病菌P. parasitica的感染 29 肆、討論 30 1. CMV介導N. tabacum cv. Samsun-NN基因靜默 30 2. NtLYM2靜默導致植株葉片出現明顯嵌紋且抑制生長 31 3. 靜默NtLYM2或NtBAK1對OPEL引發免疫反應的影響 32 4. NtCERK1在OPEL介導免疫反應的角色 34 5. 暫表現NtLYM2提升疫病菌P. parasitica之感染面積 35 6. 結語 37 伍、引用文獻 38 陸、附表 50 柒、附圖 52 捌、附錄 70 | |
| dc.language.iso | zh-TW | |
| dc.subject | CMV誘導基因靜默 | zh_TW |
| dc.subject | 菸草 | zh_TW |
| dc.subject | 植物免疫反應 | zh_TW |
| dc.subject | 疫病菌 | zh_TW |
| dc.subject | pattern-triggered immunity | en |
| dc.subject | OPEL | en |
| dc.subject | receptor-like proteins (RLPs) | en |
| dc.subject | receptor-like kinases (RLKs) | en |
| dc.subject | CMV-mediated gene silencing | en |
| dc.subject | Nicotiana tabacum | en |
| dc.subject | Phytophthora parasitica | en |
| dc.title | 探討OPEL引發植物免疫反應之機制 | zh_TW |
| dc.title | Investigating the mechanism underlying OPEL-induced plant immune responses | en |
| dc.date.schoolyear | 110-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 吳志航(Hsuan-Jung Su),葉信宏(Daniel Wei-Chung Miao),鍾嘉綾(Po-Ling Kuo),(Hao-Chung Cheng),(Ling-San Meng),(Ling-San Meng),(Ling-San Meng) | |
| dc.subject.keyword | 植物免疫反應,菸草,疫病菌,CMV誘導基因靜默, | zh_TW |
| dc.subject.keyword | OPEL,receptor-like proteins (RLPs),receptor-like kinases (RLKs),CMV-mediated gene silencing,Nicotiana tabacum,pattern-triggered immunity,Phytophthora parasitica,, | en |
| dc.relation.page | 70 | |
| dc.identifier.doi | 10.6342/NTU202200490 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2022-02-10 | |
| 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 | 2027-02-09 | - |
| 顯示於系所單位： | 植物病理與微生物學系 | |
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