請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81812完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 許如君(Ju-Chun Hsu) | |
| dc.contributor.author | Hong-Ming Chao | en |
| dc.contributor.author | 趙鴻銘 | zh_TW |
| dc.date.accessioned | 2022-11-25T03:04:09Z | - |
| dc.date.available | 2027-02-09 | |
| dc.date.copyright | 2022-02-21 | |
| dc.date.issued | 2022 | |
| dc.date.submitted | 2022-02-10 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81812 | - |
| dc.description.abstract | "秋行軍蟲 (Spodoptera frugiperda (J.E. Smith)) 為一全球重要經濟害蟲，已有超過一百個國家記錄到秋行軍蟲入侵，其危害已造成全球重大經濟損失。目前針對秋行軍蟲為害最主要防治方法仍以藥劑防治為主，而全球已有多起報導指出秋行軍蟲對多種藥劑產生抗藥性，因此本研究首先測試臺灣五個地區 (金門、花蓮、雲林、嘉義、台南) 及兩實驗室品系 (Lab-Y、Lab-P) 的秋行軍蟲，分析其對目前三種現行防治藥劑賜諾特 (spinetoram)、諾伐隆 (novaluron) 以及剋安勃 (chlorantraniliprole) 及三種防治鱗翅目害蟲的滅芬諾 (methoxyfenozide)、賽安勃 (cyantraniliprole) 及賽速安勃 (chlorantraniliprole + thiamethoxam) 的感受性現況，結果顯示賜諾特為本次實驗感受性最高之藥劑，其半致死濃度 (LC50) 及百分之90致死濃度 (LC90) 均小於3 mg/L；金門與雲林地區與室內品系相比對剋安勃的感受性，其抗性倍數 (resistance ratio) 達到10倍以上，且金門地區對二醯胺類藥劑 (diamides) 之半致死濃度均有較高之情形。三種現行防治藥劑除賜諾特外，剋安勃及諾伐隆之LC90均高於田間所推薦防治施用濃度，而三種防治鱗翅目害蟲藥劑之LC90同樣高於防治施用之濃度。進一步針對全球已知秋行軍蟲與三類殺蟲劑相關的作用標的抗性點突變，進行上述五處地點及桃園地區秋行軍蟲族群進行檢測，臺灣地區秋行軍蟲在各地均有檢測到一個乙醯膽鹼酯酶 (acetylcholinesterase) 抗性點突變 (F290V)，且花蓮、金門、台南、嘉義地區所檢測頻度達到50% 以上，而其餘兩位置A201S、G227A及發生於電壓門控鈉離子通道 (voltage-gated sodium channel) 抗性點突變T929I、L932F 及 L1014F及作用於魚尼丁受體 (ryanodine receptor) 抗性點突變I4790L及G4946E均沒有在所採樣地區被檢測到。於2021年採集自花蓮之秋行軍蟲幼蟲 (F0) 進行三作用機制點突變驗證，以現行登記濃度 (1000倍稀釋) 之硫敵克 (thiodicarb)、剋安勃 (2500倍稀釋)，再以2倍登記濃度 (400倍稀釋) 護賽寧 (flucythrinate)處理，經後72小時剋安勃死亡率達87 % 最高，硫敵克及護賽寧分別為 86 % 及67 %，各處理72小時後存活之蟲體再進行相關作用機制點突變檢測，結果僅在硫敵克處理後檢測到在乙醯膽鹼酯酶抗性點突變F290V具有 60 % 的點突變頻度。本研究最後以2019年雲林地區採集之秋行軍蟲F1及間隔一年於原地採集之FN與雲林採集帶回實驗室飼養之第九代 (F9) 之秋行軍蟲進行轉錄組定序，每個樣本獲得約6 G轉錄體，並找出現有代謝抗藥性相關顯著差異表現 (log2 FPKM ratio > 1) 基因，雲林採集F1與F9比較後共發現數條如細胞色素P450 (cytochrome P450, CYP)、穀胱甘肽S-轉移酶 (glutathione S-transferase, GST) 及羧酸酯酶 (carboxylesterase) 等基因具有顯著差異表現，雲林原地採集FN與F9比較後同樣數條如CYP、GST及二磷酸葡萄糖醛酸基轉移酶 (UDP-glucuronosyltransferase) 等顯著差異表現基因，推測這些具有顯著差異表現之代謝酵素可能與抗藥性有關。本研究結果除可提供田間防治秋行軍蟲用藥參考，並且可作為未來國內秋行軍蟲抗藥性相關研究更深入探討的基礎、抗性管理及增加秋行軍蟲防治藥劑之方向。" | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-25T03:04:09Z (GMT). No. of bitstreams: 1 U0001-0602202213404000.pdf: 2303505 bytes, checksum: ee32d0dc9338b027b1c91ff0ea973186 (MD5) Previous issue date: 2022 | en |
| dc.description.tableofcontents | "口試委員會審定書i 誌謝ii 中文摘要iii 英文摘要v 目錄vii 表次x 圖次xi 附錄次xii 壹、前言 1 貳、往昔研究 3 一、秋行軍蟲之簡介 3 (一) 秋行軍蟲之形態學及生物學 3 (二) 秋行軍蟲經濟重要性 5 (三) 秋行軍蟲全球散播及臺灣因應措施 5 (四) 秋行軍蟲防治措施 6 (五) 秋行軍蟲對殺蟲劑的抗藥性 8 二、殺蟲劑之簡介 9 (一) 硫敵克之分類及作用機制 10 (二) 護賽寧、依芬寧分類及作用機制 10 (三) 賜諾特之分類及作用機制 11 (四) 蘇力菌之分類及作用機制 12 (五) 諾伐隆之分類及作用機制 12 (六) 二醯胺類藥劑之分類及作用機制 13 (七) 滅芬諾之分類及作用機制 13 三、次世代定序 (Next Generation Sequencing, NGS) 在抗藥性研究之簡介 14 (一) DNA定序發展歷程 14 (二) 次世代定序發展與應用 14 參、材料與方法 16 一、供試蟲源採集及飼養 16 二、藥劑來源與配置 16 三、化學藥劑對秋行軍蟲感受性檢定 17 四、秋行軍蟲於作用標的抗性基因點突變檢測 18 (一) Total RNA萃取 18 (二) 反轉錄cDNA 19 (三) 聚合酶鏈鎖反應 20 (四) 利用桑格 (Sanger) 定序檢測作用位置是否有點突變情形發生 20 五、整理藥劑在不同壓力程度下秋行軍蟲基因的表現量差異..........................20 (一) RNA萃取 20 (二) 註釋結果分析 20 六、統計分析 21 肆、結果 22 一、不同地區秋行軍蟲對藥劑之感受性 22 二、田間秋行軍蟲抗性基因點突變之測定 23 三、藥劑在不同壓力下秋行軍蟲基因差異表現 24 四、GO與 KEGG 代謝途徑的比對註釋分析結果 24 伍、討論 27 一、臺灣地區秋行軍蟲對六種殺蟲劑之感受性 27 二、臺灣地區秋行軍蟲於作用標的抗性基因點突變頻度 29 三、抗藥性相關顯著表現基因分析 30 陸、結論 33 柒、引用文獻 58 捌、附錄 78 表次 表一、2019-2021年間於臺灣不同地區採集秋行軍蟲資訊 34 表二、進行室內品系秋行軍蟲對六種測試藥劑試驗之感受性結果 35 表三、進行田間品系秋行軍蟲對六種測試藥劑試驗之感受性結果 36 表四、以皮爾森相關性比較六種殺蟲劑間對秋行軍蟲田間族群抗性程度相關性 38 表五、以史比爾曼等級相關性比較秋行軍蟲於不同地區間對藥劑抗性程度相關性 39 表六、臺灣地區秋行軍蟲不同作用機制點突變基因頻度檢測結果 40 表七、秋行軍蟲驗證藥劑試驗結果 41 表八、秋行軍蟲雲林F9及FN品系之間具有顯著表現之主要代謝酵素數量及分群 42 表九、秋行軍蟲雲林F9及F1品系之間具有顯著表現之主要代謝酵素數量及分群 43 圖次 圖一、室內品系(Lab-Y) 對不同濃度賽速安處理之死亡情形 44 圖二、將 PCR 反應之含AChE1點突變基因片段產物以瓊脂凝膠進行水平電泳結果 45 圖三、秋行軍蟲與有機磷或胺基甲酸鹽類藥劑抗性相關點突變在AChE1上已知三處點突變及各田間地區品系檢測結果 46 圖四、秋行軍蟲與合成除蟲菊類藥劑抗性相關點突變在VGSC上已知三處點突變及各田間地區品系檢測結果 47 圖五、將 PCR 反應之含VGSC點突變基因片段產物以瓊脂凝膠進行水平電泳結果圖 48 圖六、將 PCR 反應之含RyR點突變基因片段產物以瓊脂凝膠進行水平電泳結果圖 49 圖七、秋行軍蟲與二醯胺類藥劑抗性相關點突變在RyR上已知兩處點突變及各田間地區品系檢測結果 50 圖八、次世代定序中秋行軍蟲具有顯著性表現基因數量 51 圖九、比較雲林F1和雲林F9具有差異表現基因之顯著GO分類圖 52 圖十、比較雲林FN和雲林F9具有差異表現基因之顯著GO分類圖 53 圖十一、GO中與現有已知抗藥性相關中表現量具有顯著性差異的秋行軍蟲基因數量 54 圖十二、比較雲林F1和雲林F9差異表現基因之顯著KEGG分類圖 55 圖十三、比較雲林FN和雲林F9差異表現基因之顯著KEGG分類圖 56 圖十四、KEGG分析結果中與現有已知抗藥性相關中表現量具有顯著性差異的秋行軍蟲基因數量 57 " | |
| dc.language.iso | zh-TW | |
| dc.subject | 轉錄體定序 | zh_TW |
| dc.subject | 點突變 | zh_TW |
| dc.subject | 秋行軍蟲 | zh_TW |
| dc.subject | 抗藥性 | zh_TW |
| dc.subject | 感受性 | zh_TW |
| dc.subject | RNA-Seq | en |
| dc.subject | Spodoptera frugiperda | en |
| dc.subject | susceptibility | en |
| dc.subject | point mutation | en |
| dc.subject | insecticide resistance | en |
| dc.title | 臺灣地區秋行軍蟲對藥劑感受性與抗藥性機制研究 | zh_TW |
| dc.title | The susceptibility to the insecticides and resistance mechanisms of Spodoptera frugiperda in Taiwan | en |
| dc.date.schoolyear | 110-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 吳文哲(Wen-Jer Wu) | |
| dc.contributor.oralexamcommittee | 戴淑美(Benny Kwok-Kan Chan),林彥伯(Chih-Yu Chiu),楊永裕 | |
| dc.subject.keyword | 秋行軍蟲,感受性,點突變,抗藥性,轉錄體定序, | zh_TW |
| dc.subject.keyword | Spodoptera frugiperda,susceptibility,point mutation,insecticide resistance,RNA-Seq, | en |
| dc.relation.page | 83 | |
| dc.identifier.doi | 10.6342/NTU202200297 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2022-02-11 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 植物醫學碩士學位學程 | zh_TW |
| dc.date.embargo-lift | 2027-02-09 | - |
| 顯示於系所單位： | 植物醫學碩士學位學程 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| U0001-0602202213404000.pdf 此日期後於網路公開 2027-02-09 | 2.25 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。