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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳昭瑩 | |
| dc.contributor.author | Ting-Li Wu | en |
| dc.contributor.author | 吳亭利 | zh_TW |
| dc.date.accessioned | 2021-06-15T13:24:36Z | - |
| dc.date.available | 2018-07-06 | |
| dc.date.copyright | 2016-07-06 | |
| dc.date.issued | 2016 | |
| dc.date.submitted | 2016-06-14 | |
| dc.identifier.citation | 行政院農委會農糧署. 2015. 2015年3月主要切花生產預測. 台灣花卉園藝 133, 8-9.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/51063 | - |
| dc.description.abstract | 本研究以次世代定序及基因微陣技術全面性分析百合灰黴病菌Botrytis elliptica侵襲百合葉片過程中，臘狀芽孢桿菌Bacillus cereus C1L透過影響那些防禦相關荷爾蒙傳訊區塊(sector)調節植物的抗病反應，以幫助鑑定單子葉植物受根圈細菌誘發的防禦荷爾蒙調控路徑，增進對單子葉作物抗病生理的瞭解。收取僅處理B. cereus C1L、僅接種百合灰黴病菌、處理B. cereus C1L後接種百合灰黴病菌、無處理無接種對照等四組百合的全RNA進行次世代定序分析，共獲得78,674個contigs，其中能比對到參照序列者占總數之35%。為鑑定B. cereus C1L誘發百合抗灰黴病過程中特有的基因差異性表現，選出各處理間變異度超過0.5，且於次世代定序與基因微陣列分析中表現趨勢一致的contigs進行文氏圖分析，發現有383個contigs表現量上升，113個contigs表現量下降，其防禦相關功能包括參與離層酸訊息路徑、調控氣孔開閉、過氧化氫相關反應、表現總穀胱甘肽硫轉移酶等，暗示離層酸傳訊路徑可能在B. cereus C1L誘導百合系統性抗灰黴病過程中扮演重要角色。為進一步探討預處理B. cereus C1L可能通過影響那些防禦傳訊區塊以強化百合對灰黴病的抗性，選取與阿拉伯芥防禦訊息路徑中關鍵基因具有最高相似度的百合同源contigs，並以其基因微陣列分析數據進行熱圖譜分析，結果顯示B. cereus C1L可能經由調控百合之茉莉酸、乙烯、離層酸傳訊區塊增進植物之系統抗病性；即時定量聚合酶連鎖反應的結果亦證實百合在抵抗灰黴病期間，B. cereus C1L預處理會促使茉莉酸傳訊區塊更加活化。此外，預處理B. cereus C1L能促進百合葉部氣孔關閉以減少百合灰黴病菌的感染，暗示離層酸傳訊區塊應參與此抗病性調控。在葉盤接種試驗中同步處理防禦相關荷爾蒙，發現同步處理茉莉酸、乙烯或離層酸可降低百合灰黴病的病徵發展，但同步處理水楊酸會使病徵輕微加劇，點明在百合灰黴病菌攻擊當下即時增加茉莉酸、乙烯或離層酸的含量可能促進百合的抗病性，暗示活化茉莉酸/乙烯及離層酸傳訊區塊在百合防禦灰黴病上具有正面貢獻。綜合文氏圖、熱圖譜、葉盤試驗及氣孔關閉比例分析之結果，推斷B. cereus C1L經由調控茉莉酸/乙烯及離層酸傳訊區塊，誘發百合對灰黴病之系統性抗病。 | zh_TW |
| dc.description.abstract | This research is to analysis the influence of Bacillus cereus C1L on what hormone signaling sectors to regulates defense response of Botrytis elliptica-infected lily by next-generation sequencing (NGS) and microarray analyses. The result may help to identify the regulation of monocot defense related hormone signaling pathway triggered by rhizobacteria and promotes the understanding of resistance physiology of monocot. Total RNAs were collected from B. cereus C1L-drenched leaves, B. elliptica-inoculated leaves, B. elliptica-inoculated leaves pre-drenched with B. cereus C1L and untreated control of lily plants for NGS analysis. Among 78,674contigs, 35% of contigs hit the reference sequences. To identify unique gene differentially expressed while triggered by B. cereus C1L on B. elliptica-inoculated lily, a criteria of variety over 0.5 of each treatment was set for selecting those contigs with expression pattern similar between NGS and customized array to create venn diagram. In this venn diagram analysis, expression of 383 contigs exhibited up-regulation while 113 contigs were down-regulated. Their functions related to plant defenses involved abscisic acid (ABA) signaling, stomatal closure regulation, hydrogen peroxide response and glutathione S-transferase, indicating that ABA signaling may play a role in lily systemic resistance against B. elliptica as triggered by B. cereus C1L. To further study about which hormone signaling pathway B. cereus C1L may utilize to strengthen resistance of lily against B. elliptica, the most similar contig homologs to the selected key gene sequences in different defense signaling pathway of Arabidopsis were selected for the analysis of heat map. Results showed that B. cereus C1L probably increase lily resistance through jasmonic acid (JA), ethylene (ET) and ABA signaling pathway. Quantitative real-time polymerase chain reaction analysis verified that pre-drench with B. cereus C1L could activate JA signaling sector upon B. elliptica attack. Besides, ABA treatment could increase stomatal closure rate and reduce the infection by B. elliptica, indicating an involvement of ABA signaling in C1L-conducted defense response. In the leaf disc assay, treatments with JA, ET or ABA at the same time of B. elliptica inoculation decreased symptom development; however, treatment with salicylic acid did not show significant effect on symptom development. These results collectively suggested that activations of JA/ET and ABA signaling sectors had positive contribution to the defense against B. elliptica. As a whole, according to results of venn diagram, heat map, leaf disc test and stomatal closure rate, B. cereus C1L–induced systemic resistance against B. elliptica in lily regulated by JA/ET and ABA signaling sectors was presumed. | en |
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| dc.description.tableofcontents | 中文摘要 II
英文摘要 IV 縮寫參照表 VI 壹、 前言 1 貳、 前人研究 3 一、 植物與環境的互動 3 1. 植物的感知行為 3 二、 植物荷爾蒙於植物對環境反應的角色 4 1. SA 傳訊區塊 5 2. JA與ET 傳訊區塊 6 3. ABA與環境反應的調控 7 4. 荷爾蒙間的交流（Cross talk） 8 5. ISR、SAR及植物能量投資之成本考量 10 三、 單子葉植物之感知與誘導抗性反應 10 四、 百合灰黴病之發生與防治 12 1. 百合之生產概況 12 2. 百合灰黴病之發生 13 3. 百合灰黴病之防治 13 五、 B. cereus C1L誘導百合抗灰黴病 14 六、 轉錄體研究現況 15 參、 材料與方法 18 一、 供試材料之製備與保存 18 1. 供試植株之製備 18 2. 供試菌株之培養與保存 18 二、 百合轉錄體之建立與分析 18 1. 試驗之處理設計 18 2. NGS之資料庫整建 19 A. 過濾病原菌序列與資料庫之選用 19 B. 覆蓋率(coverage rate)分析 19 三、 以客製化基因微陣列(Customized Microarray)分析百合基因表現量 20 1. 客製化基因微陣列之設計 20 2. RNA樣品之製備 20 3. 基因微陣列資料庫建立與分析 21 四、 NGS及基因微陣列分析之基因表現比較 21 五、 荷爾蒙傳訊區塊表現分析 22 六、 即時定量聚合酶連鎖反應 22 七、 荷爾蒙之施用測試 23 八、 C1L施用對百合氣孔關閉之影響測試 23 肆、 結果 24 一、 百合NGS資料庫分析 24 1. 轉錄體contigs組成分類 24 2. NGS contigs之GO分析 24 二、 百合NGS與基因微陣列之轉錄體分析比較 25 1. 百合轉錄體之基因微陣列分析 25 2. 百合NGS轉錄體及基因微陣列分析之一致性比較 25 3. 各處理特別表現之基因分析 25 4. C1L+BE處理特有之基因組成分析 26 5. C1L對接種BE之百合影響最高之全面性分析 26 三、 以基因微陣列分析B. cereus C1L誘導百合系統抗病性之荷爾蒙傳訊部門 27 1. SA傳訊部門 27 2. JA傳訊部門 27 3. ET傳訊部門 28 4. ABA傳訊部門 28 5. 傳訊部門間的交流分析 28 6. 以Q-RT-PCR分析C1L處理對百合抗灰黴病傳訊部門之影響 29 7. 外施荷爾蒙對百合灰黴病病徵發展之影響 29 8. 四種處理對氣孔關閉比例之影響 29 伍、 討論 30 一、 百合轉錄體的異同 30 二、 NGS及基因微陣列之表現分析 32 三、 分析方法之比較 33 四、 C1L與百合之關係建立 34 五、 C1L調控各個荷爾蒙傳訊部門影響百合灰黴病的病勢發展 35 六、 ABA及其他抗病機制在C1L影響百合抗病表現中扮演的角色 37 七、 外施荷爾蒙對百合灰黴病罹病之影響 40 八、 C1L的促進植物生長效果 40 陸、 結論 42 柒、 參考文獻 44 捌、 圖表集 51 表一、臘狀芽孢桿菌C1L菌株處理之接種百合灰黴病菌葉片基因表現上升程度最高之contigs 52 表二、臘狀芽孢桿菌C1L處理之接種百合灰黴病菌葉片基因表現下降程度最高之contigs 53 表三、臘狀芽孢桿菌C1L處理之接種百合灰黴病菌葉片茉莉酸傳訊部門各階層之contigs 54 表四、定量即時聚合酶連鎖反應使用之百合基因引子序列 55 圖一、百合轉錄體之分析流程圖 56 圖二、百合轉錄體之註解 57 圖三、百合轉錄體與資料庫資訊比對 58 圖四、臘狀芽孢桿菌C1L對感染百合灰黴病菌之葉片GO亞群基因之影響 59 圖五、次世代定序與客製化晶片之contig表現一致性分析 60 圖六、次世代定序與客製化晶片undefined contig之表一致性分析 61 圖七、Defined contigs表現之文氏圖 62 圖八、Undefined contigs之文氏圖 63 圖九、不同處理影響百合表現基因數分析 64 圖十、水楊酸傳訊部門之熱圖譜分析 65 圖十一、茉莉酸傳訊部門之熱圖譜分析 66 圖十二、乙烯傳訊部門之熱圖譜分析 67 圖十三、離層酸傳訊部門之熱圖譜分析 68 圖十四、百合茉莉酸傳訊部門基因成員之定量即時聚合酶連鎖反應分析 69 圖十六、同步施用植物荷爾蒙對百合灰黴病病斑發展之影響 (葉盤試驗) 71 圖十七、不同處理對氣孔開閉之影響 72 玖、 附錄 73 附錄一、各處理之百合葉病徵圖 74 附錄二、客製化晶片與次世代定序之表現圖譜比較分析 75 附錄三、客製化晶片與次世代定序未定義Contigs之表現圖譜比較分析。 76 附錄四、Gene ontology之生物途徑 77 附錄五、Gene ontology之細胞組件 78 附錄六、Gene ontology之分子功能 79 附錄七、預先澆灌Bacillus cereus C1L並接種Botrytis elliptica處理下特有表現之contigs 80 | |
| dc.language.iso | zh-TW | |
| dc.title | 臘狀芽孢桿菌C1L增強對灰黴病抗性之百合轉錄體分析 | zh_TW |
| dc.title | Transcriptome analysis of lily exhibiting enhanced resistance against gray mold triggered by Bacillus cereus C1L | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 104-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 鄭秋萍,黃祥恩,劉力瑜,林詩舜 | |
| dc.subject.keyword | 臘狀芽孢桿菌 C1L,百合灰黴病,轉錄體分析,基因微陣技術,茉莉酸/乙烯傳訊區塊,離層酸傳訊區塊, | zh_TW |
| dc.subject.keyword | Bacillus cereus C1L,Botrytis elliptica,transcriptome analysis,microarray,JA/ET signaling sector,ABA signaling sector, | en |
| dc.relation.page | 94 | |
| dc.identifier.doi | 10.6342/NTU201600350 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2016-06-15 | |
| 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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