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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張皓巽(Hao-Xun Chang) | |
| dc.contributor.author | Hsien-Hao Liu | en |
| dc.contributor.author | 劉軒豪 | zh_TW |
| dc.date.accessioned | 2022-11-24T03:11:35Z | - |
| dc.date.available | 2021-11-05 | |
| dc.date.available | 2022-11-24T03:11:35Z | - |
| dc.date.copyright | 2021-11-05 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-10-22 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/80643 | - |
| dc.description.abstract | "微菌核 (microsclerotia) 為真菌的特殊殘存構造之一，具繁殖能力且可承受多樣環境壓力。前人研究指出微菌核與菌核之結構和發展模式相似，並認為活性氧 (Reactive oxygen species, ROS) 為刺激微菌核與菌核分化的重要因子。然而目前活性氧刺激菌核與微菌核分化的相關研究，多以子囊菌 (Ascomycetes) 糞殼菌綱(Sordariomycetes) 中的菌核病菌 (Sclerotinia sclerotium) 、黃萎病菌 (Verticillium dahliae) 與綠殭菌 (Nomuraea rileyi) 為主，且文獻多數支持過氧化氫 (hydrogen peroxide) 為主要刺激分化的活性氧種類。關於活性氧是否在演化分歧後仍參與座殼菌綱 (Dothideomycetes) 炭腐病菌之微菌核形成，目前尚無實驗探究。本研究進行核糖核酸測序 (RNA-Sequencing)，分析炭腐病菌由菌絲至微菌核形成的四個時期 (MS0-MS3) 基因表現差異，並以基因表現趨勢分析 (cluster analysis) 與功能富集分析 (functional enrichment analysis) 發現六個基因群中的四個基因群與活性氧代謝具相關性，包括過氧化體 (peroxisome)、單加氧酶活性 (monooxygenase activity)、麩胱甘肽轉移酶 (glutathione transferase) 等。另於菌絲 (MS0) 與微菌核形成階段 (MS1-MS3) 差異表現基因分析，發現1683個上調基因與1552個下調基因之中，氧化還原和抗氧化物活性相關功能之比例皆顯著提升，其中MS1階段主要以超氧化物 (superoxide) 代謝，MS2與MS3階段則以過氧化氫代謝為主。總觀基因表現，分析結果支持活性氧參與炭腐病菌微菌核形成。抗氧化物抑制測試與活性氧染色試驗結果顯示，添加抗壞血酸 (ascorbic acid)、二硫蘇糖醇 (dithiothreitol) 與麩胱甘肽 (glutathione) 皆可抑制微菌核形成，且抑制率隨濃度上升而增加。基於50%微菌核抑制率之濃度下進行活性氧染色，發現NBT染色程度於三種抗氧化物處理相似，然而於二硫蘇糖醇處理組，DAB的染色程度較低與其他兩種抗氧化物處理具顯著差異，推測超氧化物可能為刺激微菌核形成的主要因子。另於培養基添加DETC與過氧化氫實驗中，僅於添加誘導超氧化物累積之DETC具有效刺激微菌核合成。本研究整合轉錄體與活性氧相關試驗結果，證實活性氧參與炭腐病菌微菌核形成，且發現超氧化物為刺激炭腐病菌之菌絲分化成微菌核的主要活性氧種類。" | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-24T03:11:35Z (GMT). No. of bitstreams: 1 U0001-2010202122591800.pdf: 4923114 bytes, checksum: 585705188f61cd390ceaa6be92f37915 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | "誌謝 iii 摘要 iv Abstract v 目錄 vi 表目錄 viii 圖目錄 ix 第壹章 前人研究 1 1.1 導論 1 1.2 炭腐病菌的基本特性 1 1.3 菌核 (sclerotia) 釋義 2 1.4 微菌核 (microsclerotia) 釋義 3 1.5 微菌核與菌核之比較 4 1.6 微菌核與菌核的形成－光照、溫度、養分之影響 5 1.7 活性氧 (reactive oxygen species, ROS) 與型態變化的相關性 6 1.8 ROS刺激菌核與微菌核形成 7 1.9 ROS種類之菌核與微菌核形成相關性 8 第貳章 材料與方法 10 2.1 菌株與培養條件 10 2.2 微菌核四個形成階段之純化 10 2.3 微菌核RNA萃取 11 2.4 核糖核酸定序 (RNA-Sequencing, RNA-Seq) 11 2.5 抗氧化物與微菌核之形成抑制測試 12 2.6 ROS誘導物與微菌核之形成促進測試 13 2.7 微菌核形成之H2O2與O2-染色試驗 13 第參章 結果 14 3.1 微菌核純化與轉錄體定序 14 3.2 群集分析顯示微菌核形成特定基因群與ROS具相關 15 3.3 微菌核形成之表現差異基因參與ROS代謝反應 16 3.4 抑制ROS形成能降低微菌核的形成 17 3.5 ROS種類於微菌核形成之相異性 17 3.6 ROS染色顯示O2-刺激炭腐病菌微菌核形成 18 3.7 O2-與H2O2代謝基因於炭腐病菌微菌核形成之表現 19 第肆章 討論 21 4.1 應用RNA-Seq探討微菌核形成與微菌核純化方法之建立 21 4.2 微菌核與菌核形成主要基因群與ROS代謝具相關性 22 4.3 抗氧化物抑制微菌核與菌核形成 23 4.4 O2-與H2O2於誘導微菌核與菌核形成之重要性 23 4.5 ROS代謝途徑之基因參與微菌核及菌核形成 24 參考文獻 28 表 43 圖 68 " | |
| dc.language.iso | zh-TW | |
| dc.subject | 微菌核 | zh_TW |
| dc.subject | 超氧化物 | zh_TW |
| dc.subject | 炭腐病菌 | zh_TW |
| dc.subject | 轉錄體 | zh_TW |
| dc.subject | 活性氧 | zh_TW |
| dc.subject | Microsclerotia | en |
| dc.subject | Macrophomina phaseolina | en |
| dc.subject | Reactive Oxygen Species (ROS) | en |
| dc.subject | Transcriptomics | en |
| dc.subject | Superoxide | en |
| dc.title | 整合轉錄體學與活性氧抑制實驗以探討炭腐病菌微菌核之形成 | zh_TW |
| dc.title | Integration of RNA-Seq and Reactive Oxygen Species Inhibition Assay to Study Microsclerotia Development of Macrophomina phaseolina | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 曾敏南(Hsin-Tsai Liu),林盈宏(Chih-Yang Tseng),劉瑞芬 | |
| dc.subject.keyword | 炭腐病菌,微菌核,活性氧,超氧化物,轉錄體, | zh_TW |
| dc.subject.keyword | Macrophomina phaseolina,Microsclerotia,Reactive Oxygen Species (ROS),Superoxide,Transcriptomics, | en |
| dc.relation.page | 91 | |
| dc.identifier.doi | 10.6342/NTU202103947 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-10-22 | |
| 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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