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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 鄭秋萍(Chiu-Ping Cheng) | |
| dc.contributor.author | Lu Lin | en |
| dc.contributor.author | 林露 | zh_TW |
| dc.date.accessioned | 2021-06-16T05:18:53Z | - |
| dc.date.available | 2019-09-05 | |
| dc.date.copyright | 2014-09-05 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-08-16 | |
| dc.identifier.citation | Afzal, A.A., Wood, A.J., and Lightfoot, D.A. (2008). Plant receptor-like serine threonine kinases: roles in signaling and plant defense. Mol. Plant Microbe. Interact. 21, 507-517.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/56205 | - |
| dc.description.abstract | 由細菌Ralstonia solanacearum (Rs) 引起的土壤傳播性青枯病 (bacterial wilt，簡稱 BW) 可感染全球兩百多種植物，其中包含許多重要經濟作物，且是番茄前五大病害之一。番茄品系Hawaii7996 (H7996)是目前最為穩定的青枯病抗病植物，且已發現數個抗病數量性狀基因座(quantitative trait loci，簡稱QTL)，然而，其抗病分子機制與關鍵抗病基因卻尚未知。為深入了解 H7996遺傳性質，採用全基因組解序策略並與 Heniz 1706 (H1706) 基因組進行序列比對後，結果發現核苷酸具有多型態變異並且普遍存在於各條染色體之基因片段與非基因片段。本研究針對位在 H7996第12條染色體上之抗第一演化型青枯病菌 QTL (命名為Bwr12) 進行基因搜尋與功能研究。在Bwr12區段預估的67個基因中，包含12個leucine-rich repeat (LRR)-receptor like kinase (RLKs)與LRR-receptor like protein (RLPs)，序列分析結果顯示其中多個RLKs基因的序列在H7996與H1706中具明顯差異。病毒誘導基因靜默 (virus-induced gene silencing，簡稱 VIGS) 分析所得結果發現其中數個 RLKs 被個別或共同靜默後會造成 H7996之抗性明顯降低，且病毒誘導基因表現 (virus-mediated gene over-expression，簡稱 VMGO) 分析結果更進一步顯示12g520與12g550之表現可使不抗病番茄品系 WVa700之抗性增加，顯示12g520與12g550是 H7996抗青枯病之關鍵正調控者。 Pathogen-associated molecular patterns (PAMPs)-triggered immunity (PTI) 被認為參與在抗病數量性狀位點中，根據基因靜默試驗結果得知12g520與12g550參與青枯病 PTI 反應之癒傷葡聚醣累積、 H2O2累積及 PTI 基因PTI5生合成，具有抵抗Pseudomonas syringae pv. tomato DC3000 hrcC- mutant 與Rs Pss4 hrpG- mutant 的能力。此外，蛋白質定位結果顯示12g520坐落在細胞膜。以上研究結果不但提供了番茄抗青枯病關鍵基因的重要訊息，預期亦可有助於發展基因標記，以提高抗病育種的篩選精準度與效率。 | zh_TW |
| dc.description.abstract | Bacterial wilt (BW), caused by Ralstonia solanacearum,is a highly complex and serious disease affecting a wide range of economically important crops worldwide, including tomato. Breeding for durable resistance in crops for this disease is important and urgent. Thus, comprehensive information on plant defense mechanisms would be highly desirable and necessary, but are still limited. Tomato cultivar Hawaii 7996 (H7996) is currently the most durable resistance source against BW. Despite various BW-resistance associated quantitative trait loci(QTLs) have been mapped on H7996 chromosomes, the involved molecular mechanism and gene identity remain undetermined. To gain insight into the genetic nature of H7996, we have determined its genome draft. Genome-wide sequence comparison analyses showed high sequence variations on several chromosomes between H7996 and the announced BW-susceptible cultivar Heniz 1706. In addition, we also carried out functional study on 12 putative leucine-rich repeat (LRR)-receptor like kinases (RLKs) and LRR-receptor like proteins (RLPs), which are located on one major QTL (Bwr12) associated with the H7996 resistance against Rs phylotype I strains. Comparative sequence analysis in H7996 and the Heniz1706 revealed significant sequence diversity in most of these genes. Virus-induced gene silencing (VIGS) assays suggested the involvement of a few RLKs in H7996 BW-resistance, and virus-mediated gene silencing (VMGO) assays further confirmed a positive role of 12g520 and 12g550 in H7996 BW-resistance. Pathogen-associated molecular patterns (PAMPs)-triggered immunity (PTI) is suggested to be involved in disease-resistance associated quantitative traits. Furthermore, transient gene silencing assays suggested play an important role in PTI responses including callose deposition, H2O2accumulation and marker gene PTI5 expression against Pseudomonas syringaepv. tomato DC3000 hrcC- mutant and Rs Pss4 hrpG- mutant. In addition, GFP-12g520 is localized at the plasma membrane. These results together not only provide important information on BW-resistance genes involved in H7996 defense, as well as pave the way to develop gene-based markers useful for improving efficacy and efficiency of tomato breeding programs in industry and public sectors. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T05:18:53Z (GMT). No. of bitstreams: 1 ntu-103-R00b42013-1.pdf: 6616152 bytes, checksum: 6d5142af0cb5847c33859a002c674e8b (MD5) Previous issue date: 2014 | en |
| dc.description.tableofcontents | 口試委員審定書 i
謝誌 ii 中文摘要 iv Abstract v 常用縮寫與全名對照表 vi 目錄 viii 表目錄 xii 圖目錄 xiii 附錄目錄 xiv 第一章前言 1 1. 青枯病 (Bacterial wilt, BW) 1 1.1 阿拉伯芥之青枯病反應相關研究 1 1.2 番茄之青枯病反應相關研究 2 1.3 其他植物之青枯病反應相關研究 4 2. 植物白胺酸重覆 (Leucine rich repeat, LRR) 蛋白質的特性 4 2.1 LRR receptors 結構與其功能 5 2.2 LRR receptors 與病菌之交互作用 5 3. 植物病害防禦機制 6 3.1 植物抗病反應 7 3.2 植物抗病訊息傳導 7 3.3 PAMP-triggered immunity 引發下游相關反應 8 4.研究動機 9 第二章材料與方法 10 1. 植物材料簡介 10 1.1 番茄 10 1.2 菸草 10 2. 本研究使用之細菌與其培養條件 10 3. 選殖技術 (Cloning) 11 3.1聚合酶連鎖反應 (Polymerase Chain Reaction, PCR) 11 3.2 DNA瓊脂糖凝膠電泳（agarose gel electrophoresis） 11 3.3 TOPOR質體構築 (TOPOR cloning) 12 3.4 LR 重組互換反應 (LR recombination) 12 3.5 微量抽取質體 DNA (Minipreparation of plasmid DNA) 12 3.6 DNA片段純化 (DNA purification) 13 3.7 DNA 限制酶消化水解 (DNA digestion) 13 3.8 載體與目標片段接合 (Ligation) 13 3.9大腸桿菌勝任細胞熱休克轉型作用 (Heat shock transformation) 13 3.10 電穿孔轉型作用之勝任細胞製備 (Competent cell preparation for electroporation) 14 3.11 電穿孔轉型作用 (Electroporation transformation) 14 4. 植物 DNA 萃取 (DNA extraction from plants) 14 5. 植物 DNA 全基因組解序 (Whole genome re-sequencing of plant) 15 6. 植物 RNA 萃取 (RNA extraction from plants) 16 6.1 微量抽取植物RNA 16 6.2 去除DNA 殘留 (DNase treatment) 16 7. 基因之 cDNA 釣取 17 7.1 cDNA 資料庫製備之植物材料 (plant samples of cDNA library) 17 7.2 反轉錄反應 (Reverse transcription) 17 8. 定量聚合酶連鎖反應 (quantitative polymerase chain reaction) 18 8.1 半定量 RT-PCR (Semi-quantitative RT-PCR, RT-sqPCR) 18 8.2 定量 RT-PCR (Quantitative RT-PCR, RT-qPCR) 18 9. 病毒誘導短暫性基因靜默與大量表現 (Virus-induced gene silencing [VIGS] and virus-mediated gene overexpression [VMGO]) 19 9.1 VIGS 19 9.2 VMGO 20 10. 番茄之青枯病反應檢測 (Tomato BW assay) 20 11. 番茄 PTI 防禦反應分析 (PTI defense response in tomato) 21 11.1 病菌處理 (Pathogen treatments) 21 11.2 癒傷葡聚醣累積分析 (Callose deposition assay) 21 11.3 過氧化物檢測 (ROS accumulation assay) 21 11.4 H2O2累積檢測 (H2O2 accumulation assay) 22 11.5 PTI5轉錄表現檢測 (PTI5 expression assay) 22 12. 青枯病菌感染後番茄基因轉錄之檢測 (Analysis of tomato gene expression after Rs inoculation) 22 13. 蛋白質在菸草葉片細胞之定位分析 (Sub-cellular localization in tobacco) 23 13.1 載體構築 23 13.2 蛋白質定位分析與細胞質壁分離 23 第三章結果 25 1. 番茄Hawaii 7996 (H7996) 全基因組草圖 25 2. 番茄Bwr12區段內基因群之功能歸類與序列比對分析 26 3. 以短暫基因靜默策略分析Bwr12相關 RLKs/RLPs 在H7996 青枯病抗性之角色 27 4. 以短暫基因大量表現策略分析12g520、12g550及12g530在 H7996青枯病抗性之角色 28 5. 番茄 PTI 反應分析系統之建立 29 6. 12g520與12g550之表現量對番茄 PTI 反應的影響 31 7. 接種青枯病菌後之12g520與12g550表現 31 8. 12g520與12g550蛋白質產物在植物細胞中之座落位置 32 第四章討論 33 1. 抗青枯病番茄 H7996之數條染色體具有高度序列差異性 33 2. 青枯病菌誘發之番茄 PTI 反應 34 3. 數個Bwr12 區段之 RLK 基因參與 H7996對 phylotype I 青枯病菌之抗性 36 4. 12g520參與 H7996青枯病抗性之可能機制 37 5. 12g550參與 H7996青枯病抗性之可能機制 38 6. 12g720/740參與 H7996青枯病抗性 39 7. H7996 Bwr12之訊息傳導 39 8. 結語 40 第五章參考文獻 41 第六章附錄 49 | |
| dc.language.iso | zh-TW | |
| dc.title | 番茄抗青枯病品系Hawaii7996之解序與其抗第一演化型青枯病菌基因座Bwr12之研究 | zh_TW |
| dc.title | Genome draft of bacterial wilt resistant tomato Hawaii7996 and study of the resistance locus Bwr12 against Ralstonia solanacearum phylotype I strains | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 劉瑞芬(Ruey-Fen Liou),鄭貽生(Yi-Sheng Cheng),張英?(Ing-Feng Chang),陳逸然(Yet-Ran Chen) | |
| dc.subject.keyword | 青枯病,青枯病菌,數量性狀位點,H7996,LRR,RLK,PTI, | zh_TW |
| dc.subject.keyword | Bacterial wilt,Ralstonia solanacearum,QTL,Hawaii7996,LRR,RLK,PTI, | en |
| dc.relation.page | 108 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2014-08-17 | |
| 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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