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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳昭瑩(Chao-Ying Chen) | |
dc.contributor.author | Nai-Hua Ye | en |
dc.contributor.author | 葉乃樺 | zh_TW |
dc.date.accessioned | 2021-06-17T03:35:45Z | - |
dc.date.available | 2021-03-02 | |
dc.date.copyright | 2018-03-02 | |
dc.date.issued | 2018 | |
dc.date.submitted | 2018-02-12 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/69952 | - |
dc.description.abstract | 植物第二型富含甘胺酸蛋白LsGRP1對於葵百合防禦病原菌上有所貢獻,本研究進一步檢視LsGRP1如何牽涉於植物防禦反應。於LsGRP1轉基因阿拉伯芥上接種十字花科炭疽病菌Colletotrichum higginsianum及番茄細菌性斑點病菌Pseudomonas syringae pv. tomato DC3000 (Pst DC3000),顯示LsGRP1轉基因阿拉伯芥對病原菌具有比野生型植株較佳的防禦能力。利用第三型分泌系統缺失之Pst hrcC-突變株以及細菌病原相關分子模式flg22、真菌病原相關分子模式chitohexaose,證明LsGRP1轉基因阿拉伯芥顯著地誘發較高量的活性氧物質及癒傷葡聚醣(胼胝质)沉積,並增加呼吸爆發氧化酶基因表現;鈣離子依賴性蛋白激酶誘導性基因表現在LsGRP1轉基因阿拉伯芥上亦些微提升,顯示LsGRP1會使植株提升病原相關分子模式觸發免疫反應 (pathogen-associated molecular pattern-triggered immunity) 。除此之外,在Avr效應子 - AvrRpt2以及AvrRpm1的觸發下,LsGRP1轉基因阿拉伯芥具有比野生型植株更強的過敏性反應,顯示LsGRP1亦增強植物的效應子觸發免疫反應 (effector-triggered immunity),證實LsGRP1可以透過多方面機制來提升植物免疫力。本研究中還觀察到LsGRP1轉基因阿拉伯芥的生長增強現象,並透過觀察LsGRP1-EGFP的次細胞定位,推測LsGRP1可能在葉綠體上參與植物生長和防禦的調控機制。 | zh_TW |
dc.description.abstract | LsGRP1, a plant class II glycine-rich protein (GRP), contributes to the defense of Lilium cv. Star Gazer against pathogen attack. How LsGRP1 might involve in plant defense was investigated in the study. Firstly, the inoculation with Colletotrichum higginsianum and Pseudomonas syringae pv. tomato DC3000 showed that LsGRP1-transgenic Arabidopsis thaliana exhibited stronger defense against pathogens than the wild-type plants. By using Pst hrcC– mutant (type III secretion mutant), bacterial pathogen-associated molecular pattern (PAMP) flg22 and fungal PAMP chitohexaose, higher levels of reactive oxygen species (ROS), callose and respiratory burst oxidase homologs D (AtrbohD) gene expression were shown in LsGRP1-transgenic Arabidopsis. In addition, calcium-dependent protein kinase-induced gene expression was slightly enhanced in LsGRP1-transgenic Arabidopsis. These results reveal that the presence of LsGRP1 conducts stronger PAMP-triggered immunity in plants. Besides, the LsGRP1-transgenic Arabidopsis showed more severe hypersensitive responses than wild-type plants after triggered by Avr proteins, AvrRpt2 and AvrRpm1, demonstrating the enhancement of effector-triggered immunity mediated by LsGRP1. Thus, LsGRP1 would enhance plant immunity through different aspects. In addition, the enhanced growth phenomenon in LsGRP1-transgenic Arabidopsis and subcellular localization of LsGRP1-EGFP revealed that LsGRP1 might involve in the regulation of plant growth and defense occurring in chloroplasts. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T03:35:45Z (GMT). No. of bitstreams: 1 ntu-107-R04633007-1.pdf: 3452386 bytes, checksum: 2eb86b7a704757d05795a2dfa745fe9b (MD5) Previous issue date: 2018 | en |
dc.description.tableofcontents | 壹、中文摘要 i
貳、英文摘要 ii 參、前言 1 肆、前人研究 3 一、植物先天免疫反應 (Plant innate immunity) 3 植物荷爾蒙訊息介導之先天免疫反應 6 二、植物富含甘胺酸蛋白 6 三、植物第二型富含甘胺酸蛋白參與植物防禦反應 8 四、水楊酸誘導葵百合防禦相關蛋白LsGRP1 8 伍、材料與方法 10 一、植物栽培 10 二、病原菌培養與接種原製備 10 三、細菌與真菌病原分子模式製備 10 四、病原細菌回收與計量 11 五、癒傷葡聚醣之植物組織染色法 11 六、ROS (H2O2) 之植物組織染色法 11 七、植物RNA萃取 12 八、植物全基因體核酸萃取 12 九、反轉錄-即時聚合酶定量連鎖反應 (Reverse transcription-quantitative PCR, RT-qPCR) 12 十、構築表現病原效應子之載體 13 1. 病原菌效應子基因增幅 13 2. 目標DNA片段膠體回收及純化 13 3. T-DNA載體構築 13 十一、構築表現病原效應子之載體 14 1. 大腸桿菌勝任細胞製備 14 2. 大腸桿菌熱休克轉形 14 3. 大腸桿菌轉形株篩選 14 4. 抽取大腸桿菌質體 14 十二、構築表現病原效應子之農桿菌 15 1. 農桿菌勝任細胞製備 15 2. 農桿菌電穿孔法 15 十三、農桿菌注入法 15 十四、植物死亡細胞Evans blue染色及量化方法 16 十五、測量阿拉伯芥幼苗之地上部生物量 16 十六、測量阿拉伯芥幼苗之根部長度 16 十七、LsGRP1-EGFP於轉基因阿拉伯芥上之定位觀察 17 十八、統計方法 17 陸、結果 18 一、檢測LsGRP1轉基因植物上的LsGRP1基因表現量 18 二、表現LsGRP1會影響十字花科炭疽病於阿拉伯芥上的感染進程 18 三、表現LsGRP1提升阿拉伯芥對番茄細菌性斑點病菌DC3000之抗性 18 四、表現LsGRP1會造成第三型分泌系統缺失之番茄細菌性斑點病菌在阿拉伯芥上的增殖量減少 19 五、表現LsGRP1會影響細菌與真菌病原分子模式誘發之癒傷葡聚醣累積 19 六、表現LsGRP1會影響細菌與真菌病原分子模式誘發之活性氧物質累積 19 七、表現LsGRP1不會影響阿拉伯芥PTI相關抗性基因之基礎表現量 20 八、表現LsGRP1會影響細菌與真菌病原分子模式誘發之活性氧物質生合成基因表現 20 九、表現LsGRP1會輕微影響鈣離子/鈣調蛋白依賴性激酶傳訊路徑誘導之基因表現 20 十、表現LsGRP1提升阿拉伯芥上病原效應子誘發之過敏性反應 21 十一、 表現LsGRP1對於阿拉伯芥幼苗生長之影響 22 十二、在持續高量表現LsGRP1-EGFP之阿拉伯芥上LsGRP1之次細胞定位 22 柒、討論 23 捌、參考文獻 28 玖、圖表集 37 表一、用以增幅基因之引子 38 表二、即時聚合酶鏈鎖反應使用之引子 39 圖一、以RT-qPCR檢查LsGRP1轉基因阿拉伯芥的葉片內之LsGRP1 基因表現 40 圖二、利用即時聚合酶鏈鎖反應技術檢測野生型以及LsGRP1轉基因阿拉伯芥上的真菌病原Colletotrichum higginsianum XL1-4之生物量 41 圖三、Colletotrichum higginsianum XL1-4於野生型以及LsGRP1轉基因阿拉伯芥所造成的葉部病徵 42 圖四、比較自野生型以及LsGRP1轉基因阿拉伯芥所回收之病原細菌Pseudomonas syringae pv. tomato DC3000族群量 43 圖五、比較自野生型以及LsGRP1轉基因阿拉伯芥所回收之Pseudomonas syringae pv. tomato hrcC-族群量 44 圖六、以細菌性以及真菌性病原菌相關分子模式觸發野生型以及LsGRP1轉基因阿拉伯芥累積癒傷葡聚糖 46 圖七、以細菌性以及真菌性病原菌相關分子模式觸發野生型以及LsGRP1轉基因阿拉伯芥葉片累積活性氧物質 47 圖八、AtrbohD基因以及PHI-1基因在未處理組之野生型以及LsGRP1轉基因阿拉伯芥葉片的相對表現量 48 圖九、AtrbohD基因在經flg22處理之野生型以及LsGRP1轉基因阿拉伯芥葉片的相對表現量 49 圖十、AtrbohD基因在經chitohexaose處理之野生型以及LsGRP1轉基因阿拉伯芥葉片的相對表現量 50 圖十一、PHI-1基因在經flg22處理之野生型以及LsGRP1轉基因阿拉伯芥葉片的相對表現量 51 圖十二、PHI-1基因在經chitohexaose處理之野生型以及LsGRP1轉基因阿拉伯芥葉片的相對表現量 52 圖十三、比較接種Pseudomonas syringae pv. tomato DC3000 avrRpt2菌株在野生型以及LsGRP1轉基因阿拉伯芥葉片上引起的過敏性反應 53 圖十四、以反轉錄PCR檢測農桿菌介導AvrRpt2、AvrRpm1基因在野生型以及LsGRP1轉基因阿拉伯芥葉片中的暫時性表現 55 圖十六、比較野生型以及LsGRP1轉基因阿拉伯芥幼苗的生長 58 圖十七、LsGRP1-EGFP在經細菌性或真菌性病原菌相關分子模式處理之阿拉伯芥葉組織的次細胞定位 59 拾、附錄 60 附錄一、比較在以農桿菌注入法表現LsGRP1或其缺失性突變的阿拉伯芥葉片中的十字花科炭疽病菌感染量 61 附錄二、比較AtIP40表現基因AT5G10770在野生型以及LsGRP1轉基因阿拉伯芥葉片中的相對表現量 62 附錄三、PAMPs處理對LsGRP1交互作用複合體內的蛋白組成之影響 64 附錄四、附圖三之材料方法 66 | |
dc.language.iso | zh-TW | |
dc.title | 利用LsGRP1轉基因阿拉伯芥探討其於植物免疫之角色 | zh_TW |
dc.title | The roles of transgenic Arabidopsis-expressing LsGRP1 in plant immunity | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 鄭秋萍,賴爾?,黃秀珍,路幼妍 | |
dc.subject.keyword | LsGRP1,植物第二型富含甘胺酸蛋白,病原相關分子模式觸發免疫反應,效應子觸發免疫反應, | zh_TW |
dc.subject.keyword | LsGRP1,plant class II glycine-rich protein,pathogen-associated molecular pattern-triggered immunity,effector-triggered immunity, | en |
dc.relation.page | 67 | |
dc.identifier.doi | 10.6342/NTU201800532 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2018-02-12 | |
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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