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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 | Chia-Yen Lu | en |
dc.contributor.author | 呂佳燕 | zh_TW |
dc.date.accessioned | 2021-06-16T05:49:42Z | - |
dc.date.available | 2017-08-25 | |
dc.date.copyright | 2014-08-25 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-08-08 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/56807 | - |
dc.description.abstract | 植物促生根圈細菌(plant growth-promoting rhizobacteria, PGPR)是指能群聚於植物根圈之益生細菌,有些能促進植物生長,甚至可誘發植物產生系統抗病性。本實驗室前人自太魯閣國家公園之臺灣百合根圈分離出一PGPR菌株—臘狀芽孢桿菌C1L,已證實澆灌其菌液於根圈土壤,能有效降低百合灰黴病及玉米葉枯病的罹病程度。本研究欲了解玉米與臘狀芽孢桿菌C1L之間的交互作用,從實驗室前人製備之C1L轉位子突變株庫中,挑選喪失誘導玉米產生系統性抗病能力之突變基因-磷酸轉移酶系統(phosphotransferase system, PTS)之葡萄糖專一型IIABC組成的解碼基因ptsG作為研究對象,探討其在細菌與玉米交互作用中所扮演的角色。而植物行光合作用之部分產物會以根分泌物的形式釋放到土壤中,為植物與根圈微生物溝通的一種形式,透過C1L野生型菌株、ptsG突變株及ptsG(pLKptsG)互補株證明ptsG基因參與C1L對玉米根分泌物之趨向行為,以及在玉米根表的增殖數量。進一步藉由螢光報導系統分析證明ptsG基因之表現量受玉米根分泌物所調控,其中之有效成分應為多種醣類。由於破壞ptsG基因會減低C1L攝取葡萄糖的能力,也導致揮發性二次代謝物3-羥基-2丁酮(acetoin)及2,3-丁二醇(2,3-butanediol)產量減少,並經由溫室試驗證明此二揮發性有機化合物可作為玉米ISR之誘導分子。故而推斷臘狀芽孢桿菌C1L之ptsG基因表現量會受玉米根分泌物調節繼而正向影響趨向性、根部群聚能力及二次代謝物生成,以誘導玉米之系統抗病性。 | zh_TW |
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dc.description.tableofcontents | 目錄
壹、中文摘要 1 貳、英文摘要 2 肆、前人研究 5 一、植物促生根圈細菌 (Plant growth-promoting rhizobacteria, PGPR) 5 二、根圈之生物防治菌 (Biocontrol rhizobacteria) 6 三、玉米與玉米葉枯病 (southern corn leaf blight) 6 四、植物根分泌物 (Root exudates) 7 五、磷酸轉移酶系統 (Phosphotransferase system, PTS) 8 六、磷酸轉移酶系統之ptsG調控機制 9 七、細菌趨向性(chemotaxis)之調控機制 10 八、微生物揮發性有機化合物(volatile organic compounds, VOCs)對植物之作用 11 伍、材料與方法 13 一、供試菌株與載體 13 1. 供試細菌菌株之培養及保存 13 2. 供試真菌菌株之培養及保存 13 二、突變株ptsG及互補株ptsG(pLKptsG)特性分析 14 1. 葡萄糖利用試驗 14 2. 玉米根分泌物趨向性測試 15 3. 玉米根部表生性測定 16 4. 玉米ISR誘導能力測試 16 5. 內孢子生成能力測試 17 三、菌株間ptsG表現量之差異 18 1. 細菌RNA抽取 18 2. 反轉錄-聚合酶連鎖反應 18 四、啟動子活性誘導試驗 19 1. 綠色螢光報導基因質體構築 19 2. 利用螢光顯微鏡觀察C1L綠色螢光報導基因之表現 23 五、玉米根分泌物組成對C1L之影響 24 1. 趨向性測試 24 2. 醣類代謝產物之誘導 25 六、2,3-丁二醇及3-羥基-2-丁酮 26 1. 誘導玉米ISR功能性測試 26 2. 揮發性有機化合物對玉米葉枯病菌之抑菌試驗 27 七、統計分析方法 27 陸、結果 28 一、ptsG突變株攝取葡萄糖效率較C1L野生株低 28 二、ptsG突變株對玉米根分泌物之趨向程度降低 28 三、ptsG突變株於玉米根部表生能力下降 29 四、ptsG參予C1L菌株誘導玉米ISR現象 29 五、ptsG突變株喪失內孢子之生成能力 30 六、ptsG訊息RNA表現量分析 30 七、玉米根分泌物誘導C1L ptsG基因表現試驗 30 1. 綠色螢光報導基因系統載體構築 31 2. 玉米根分泌物及其醣類組成誘導C1L ptsG基因表現 31 八、玉米根分泌物組成對C1L之趨向性測試 31 九、玉米根分泌物組成誘導生成之C1L次級代謝產物 32 十、2,3-丁二醇及3-羥基-2-丁酮可抑制玉米葉枯病之病徵發展 33 1. 玉米抗性誘導能力試驗 33 2. 2,3-丁二醇及3-羥基-2-丁酮對玉米葉枯病菌之抑菌試驗 34 十一、ptsG基因影響C1L 次級代謝產物3-羥基-2-丁酮及2, 3-丁二醇之生成 34 柒、討論 35 捌、總結 41 玖、參考文獻 42 壹拾、圖表集 52 表一、供試菌株 53 表二、供試載體 54 表三、引子 55 圖一、C1L野生株、ptsG突變株及ptsG互補株之葡萄糖利用能力 56 圖二、C1L野生株、ptsG突變株及ptsG互補株對玉米根分泌物之趨向性 57 圖三、C1L野生株、ptsG突變株及ptsG互補株於玉米根部表面群聚之菌量 58 圖四、玉米葉枯病之罹病指數 59 圖五、臘狀芽孢桿菌C1L野生株、ptsG突變株及ptsG互補株誘導玉米ISR能力 60 圖六、C1L野生株、ptsG突變株及ptsG互補株之內孢子產生能力 61 圖七、C1L ptsG基因表現之半定量RT-PCR分析 62 圖八、pAD123及pAD123-PptsG圖譜 63 圖九、確認電轉形株C1L(pAD123)及C1L(pAD123-PptsG) 64 圖十、玉米根分泌物誘導ptsG啟動子之綠色螢光報導基因表現 65 圖十一、以綠色螢光報導系統分析ptsG啟動子之活化條件 66 圖十二、C1L和ptsG突變株菌株對根分泌物醣類組成之趨向性反應 67 圖十三、以薄層層析法檢測C1L受醣類誘導生成之代謝產物 68 圖十四、以氣相層析/質譜分析C1L培養上清液成分 69 圖十五、C1L培養上清液之氣相層析滯留時間第3.51分鐘之質譜圖譜 70 圖十六、C1L培養上清液之氣相層析滯留時間第4.59分鐘之質譜圖譜 71 圖十七、3-羥基-2-丁酮及2,3-丁二醇可誘導玉米表現系統抗病性 72 圖十八、3-羥基-2-丁酮及2,3-丁二醇對玉米葉枯病菌生長之抑菌分析 73 圖十九、C1L野生株、ptsG突變株所產生2, 3-丁二醇之薄層層析 74 圖二十、C1L野生株及ptsG突變株所產生3-羥基-2-丁酮及2,3-丁二醇之氣相層析/質譜分析 76 壹拾壹、附錄 77 附錄一、磷酸轉移酶系統模式圖 78 附錄二、玉米根分泌物成分 79 | |
dc.language.iso | zh-TW | |
dc.title | 臘狀芽孢桿菌C1L之磷酸轉移酶系統ptsG基因參與根分泌物調節其對玉米系統抗病性之誘導能力 | zh_TW |
dc.title | The ptsG gene participating in root exudates-mediated
systemic disease resistance-inducing ability of Bacillus cereus C1L in maize | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 李永安(Yung-An Lee),吳蕙芬,林乃君,黃祥恩 | |
dc.subject.keyword | 臘狀芽孢桿菌,植物促生根圈細菌,磷酸轉移?系統,葡萄糖專一型IIABC組成解碼基因ptsG,玉米,根分泌物,誘導系統抗病性, | zh_TW |
dc.subject.keyword | Bacillus cereus C1L,plant growth-promoting rhizobacteria,phosphotransferase system,glucose-specific IIABC component ptsG,maize,root exudates,induced systemic resistance, | en |
dc.relation.page | 79 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2014-08-08 | |
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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