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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 陳昭瑩(Chao-Ying Chen) | |
dc.contributor.author | Ting-Wei Chen | en |
dc.contributor.author | 陳廷瑋 | zh_TW |
dc.date.accessioned | 2021-06-17T00:47:15Z | - |
dc.date.available | 2022-02-10 | |
dc.date.copyright | 2020-02-10 | |
dc.date.issued | 2020 | |
dc.date.submitted | 2020-02-05 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/66627 | - |
dc.description.abstract | 植物在受到病原菌等外在刺激時會發生許多生理上的變化,其訊息傳遞的過程常受到植物荷爾蒙的調控,已知水楊酸、茉莉酸、乙烯、離層酸等都可能對植物的抗性造成影響,而本實驗室於過去的研究中,也得知了水楊酸的處理會提升葵百合對灰黴病菌(Botrytis elliptica)的抗性並增加防禦相關蛋白LsGRP1的表現。為了進一步得知葵百合受到B. elliptica感染時各種植物荷爾蒙所扮演的角色,利用實驗室前人所建構的葵百合轉錄體資料庫,根據與荷爾蒙相關基因相似度高的contigs設計引子,自葵百合cDNA中選殖300~700鹼基對的序列構築至菸草脆裂病毒(tobacco rattle virus, TRV)載體,以用於病毒誘導基因靜默(virus-induced gene silencing, VIGS)試驗。經由在葵百合中ICS1、ETR1、MPK6及EIN3同源性序列的VIGS及接種測試,可於LsICS1 (AtICS1 homolog)、LsETR1 (AtETR1 homolog)、LsMPK6 (AtMPK6 homolog)及LsEIN3 (AtEIN3 homolog)靜默的植株上觀察到對B. elliptica之感病性有提升的現象,其中LsICS1及LsEIN3靜默亦會降低接種前百合葉片中的LsGRP1表現量。另一方面,在阿拉伯芥LsGRP1轉殖株上的實驗顯示AtETR1和AtMPK6靜默並不影響LsGRP1介導之阿拉伯芥對Botrytis cinerea的抗性,而AtICS1靜默的LsGRP1-7植株則抗性明顯下降。綜合這些結果推知LsGRP1有助於百合對B. elliptica的抗性,其在百合中的表現需要水楊酸及乙烯訊息傳遞路徑共同調控;並推測在阿拉伯芥上,LsGRP1介導的抗病性需AtICS1所導致的水楊酸累積。 | zh_TW |
dc.description.abstract | Perception of pathogen by plants triggers metabolic reprograming in response to pathogens, in which phytohormones are involved in signal transduction. In previous report, it is known that salicylic acid treatment improves resistance against Botrytis elliptica and increases expression of LsGRP1 in Lilium cultivar Star Gazer. To understand the role of phytohormones in the interaction between Lilium ‘Star Gazer’ and B. elliptica, sequences of length about 300 bp to 700 bp derived from putative ICS1, ETR1, MPK6 and EIN3 of Lilium ‘Star Gazer’ were amplified and constructed into tobacco rattle virus (TRV) vector for virus-induced gene silencing (VIGS) assay. Based on the reports in Arabidopsis and the transcriptome database of Lilium ‘Star Gazer’, LsICS1 (AtICS1 homolog), LsETR1 (AtETR1 homolog), LsMPK6 (AtMPK6 homolog) and LsEIN3 (AtEIN3 homolog) were selected for the VIGS assay in Lilium ‘Star Gazer’. The results showed all these VIGS constructs conducted a higher susceptibility to B. elliptica, and silencing of LsICS1 and LsEIN3 result in lower LsGRP1 expression level. In Arabidopsis, silencing of AtETR1 and AtMPK6 didn’t alter the resistance to Botrytis cinerea in LsGRP1 transgenic plants whereas silencing of AtICS1 compromised LsGRP1-mediated resistance against B. cinerea. These results suggest that expression of LsGRP1 in Lilium depends on both salicylic acid and ethylene signaling pathways and salicylic acid accumulation is essential for LsGRP1-mediated resistance against B. cinerea in Arabidopsis. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T00:47:15Z (GMT). No. of bitstreams: 1 ntu-109-R05633011-1.pdf: 8974336 bytes, checksum: 88f2123c58c6cad1959e000c308356be (MD5) Previous issue date: 2020 | en |
dc.description.tableofcontents | 壹、 中文摘要 1
貳、 英文摘要 2 參、 前言 1 肆、 前人研究 3 一、百合 3 二、百合灰黴病 4 三、植物荷爾蒙訊息傳遞路徑與植物對灰黴病之防禦反應 5 四、病毒誘導基因靜默 7 伍、 材料與方法 9 一、供試植物之栽培 9 1. 栽培介質調配 9 2. 百合的栽培 9 3. 阿拉伯芥的栽培 9 二、供試微生物之培養、保存與接種源製備 10 1. 供試微生物之培養及保存 10 2. 供試真菌之接種源製備 10 三、病毒誘導基因靜默 11 1. 病毒誘導基因靜默用片段的選殖 11 2. 農桿菌介導病毒誘導基因靜默 12 四、水楊酸之預處理 13 五、植物抗性之分析 13 六、即時定量聚合酶連鎖反應 14 七、實驗數據的統計分析 15 陸、 結果 16 一、比對與選擇百合防禦荷爾蒙相關基因標的 16 二、百合葉中誘導基因靜默之病毒載體含量偵測及基因靜默效率測定 17 三、預處理水楊酸對百合抗病性及荷爾蒙訊息傳遞路徑基因表現的影響 18 四、靜默防禦基因LsGRP1會使百合的水楊酸及茉莉酸路徑下游LsWRKY70及LsPDF2.2基因表現量降低 19 五、靜默水楊酸生合成基因LsICS1抑制百合灰黴病抗性,影響水楊酸與茉莉酸訊息路徑基因以及LsGRP1表現 19 六、靜默乙烯受體基因LsETR1會降低百合對灰黴病的抗性 20 七、靜默百合乙烯訊息路徑相關轉錄因子LsEIN3會降低百合對灰黴病的抗性 20 八、靜默百合MAPK傳訊基因LsMPK6會降低百合對灰黴病的抗性 20 九、靜默水楊酸生合成基因AtICS1會損害LsGRP1介導的阿拉伯芥灰黴病抗性 21 柒、 討論 22 一、百合中水楊酸訊息傳遞路徑與防禦基因LsGRP1表現的調控 22 二、百合中乙烯訊息傳遞路徑與LsGRP1表現的調控 23 三、百合中MAPK傳訊與LsGRP1表現的調控 24 四、百合中不同荷爾蒙訊息傳遞路徑間的交互作用 24 五、荷爾蒙訊息傳遞路徑與LsGRP1介導抗性 26 六、農桿菌介導基因靜默技術的最佳化 28 七、總結 28 捌、 參考文獻 30 玖、 圖表集 39 表一、選殖百合推定植物荷爾蒙訊息傳遞路徑相關基因片段之引子 40 表二、檢測百合目標基因表現之即時定量反轉錄聚合酶鏈鎖反應引子 41 表三:選殖阿拉伯芥的植物荷爾蒙訊息傳遞路徑相關基因之引子 42 表四、檢測阿拉伯芥目標基因表現之即時定量反轉錄聚合酶鏈鎖反應引子 43 表五、量化病原菌及病毒載體之即時定量聚合酶鏈鎖反應引子 44 圖一、阿拉伯芥ICS1與百合同源基因的胺基酸序列排比分析 45 圖二、阿拉伯芥ICS1與百合同源基因的互補核酸序列排比分析 48 圖三、阿拉伯芥ETR1與百合同源基因的胺基酸序列排比分析 49 圖四、阿拉伯芥ETR1與百合同源基因的互補核酸序列排比分析 51 圖五、阿拉伯芥EIN3與百合同源基因的胺基酸序列排比分析 52 圖六、阿拉伯芥EIN3與百合同源基因的互補核酸序列排比分析 54 圖八、阿拉伯芥MPK6與百合同源基因的互補核酸序列排比分析 57 圖九、病毒誘導基因靜默載體pTRV2示意圖及其於百合中基因靜默效果測試 59 圖十、水楊酸處理促進百合對灰黴病的抗性並影響荷爾蒙訊息傳遞路徑相關基因表現 62 圖十一、靜默LsGRP1基因降低百合對灰黴病的抗性並影響荷爾蒙訊息傳遞路徑相關基因表現 65 圖十二、靜默LsICS1基因降低百合對灰黴病的抗性並影響荷爾蒙訊息傳遞路徑相關基因表現 68 圖十三、靜默LsETR1基因降低百合對灰黴病的抗性並影響荷爾蒙訊息傳遞路徑相關基因表現 71 圖十四、靜默LsEIN3基因降低百合對灰黴病的抗性並影響荷爾蒙訊息傳遞路徑相關基因表現 74 圖十五、靜默LsMPK6基因降低百合對灰黴病的抗性並影響荷爾蒙訊息傳遞路徑相關基因表現 77 圖十六、靜默阿拉伯芥LsGRP1轉殖株的荷爾蒙訊息傳遞路徑相關基因對灰黴病抗性的影響 78 圖十七、植物荷爾蒙相關基因間轉錄調控及LsGRP1介導灰黴病抗性總覽 80 | |
dc.language.iso | zh-TW | |
dc.title | 植物荷爾蒙訊息傳遞路徑在葵百合LsGRP1介導植物防禦灰黴病之角色 | zh_TW |
dc.title | The roles of phytohormone signaling pathways in LsGRP1-mediated plant defense against gray mold | en |
dc.type | Thesis | |
dc.date.schoolyear | 108-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 沈偉強,鄭秋萍,葉信宏,路幼妍 | |
dc.subject.keyword | 植物荷爾蒙,植物防禦路徑,Botrytis elliptica,LsGRP1,葵百合,病毒誘導基因靜默,Arabidopsis,Botrytis cinerea, | zh_TW |
dc.subject.keyword | Phytohormones,plant defense pathway,Botrytis elliptica,LsGRP1,Lilium ‘Star Gazer’,virus-induced gene silencing (VIGS),Arabidopsis,Botrytis cinerea, | en |
dc.relation.page | 80 | |
dc.identifier.doi | 10.6342/NTU202000350 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2020-02-05 | |
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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