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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 柯俊成(Chiun-Cheng Ko) | |
| dc.contributor.author | Yung-Chang Hung | en |
| dc.contributor.author | 洪永昌 | zh_TW |
| dc.date.accessioned | 2021-06-13T16:28:24Z | - |
| dc.date.available | 2008-07-26 | |
| dc.date.copyright | 2005-07-26 | |
| dc.date.issued | 2005 | |
| dc.date.submitted | 2005-07-14 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/38230 | - |
| dc.description.abstract | 煙草粉蝨為一種群 (species complex)，自命名以來，迄今於世界各地已有 24 個以上的生物小種 (biotype)，而生物小種間並無明顯的形態特徵可供區分，隨著分子生物技術的發展，應用分子標示來區分煙草粉蝨生物小種的研究日益增加。本研究主要根據 DNA 逢機增幅多態型聚合酶連鎖反應 (RAPD-PCR) 及粒線體 COI 序列為基礎，開發能有效鑑別煙草粉蝨 A、B、Q、Nauru、AN 及 S 生物小種之專一性引子組。結果顯示，逢機引子 OPA15 及 OPO18 可分別針對煙草粉蝨 A 及 Q 生物小種，增幅出特異性片段，依據此片段所設計之引子組 BaAF/BaAR 及 BaQF/BaQR，則可分別針對煙草粉蝨 A 及 Q 生物小種增幅出具鑑別力之 DNA 片段；此外，4 個正向引子 BaBF、BaNaF、BaANF 及 BaSF 則根據比對不同煙草粉蝨生物小種之粒線體 COI 序列設計，分別與粒線體 COI 之反向引子 L2-N-3014 配合，可針對煙草粉蝨 B、Na、AN 及 S 生物小種增幅出專一性片段。另一方面，臺灣地區所採集之 202 個煙草粉蝨族群，利用二級內共生菌之 16S rDNA 及Wolbachia 之 wsp 基因進行內共生菌的偵測，結果顯示，55.4% 的族群受二級內共生菌的感染，38.1% 的族群受 Wolbachia 的感染，再經由類緣關係分析，感染臺灣地區煙草粉蝨之二級內共生菌皆為 Proteobacteria γ-3 亞綱的種類，而 Wolbachia 則屬於 B 群的 Con/Rug 亞群的種類。由各生物小種感染之比例顯示，此二類內共生菌之分布與生物小種有密切的相關，然而影響其分布的因素仍需後續的研究以進一步釐清。 | zh_TW |
| dc.description.abstract | Bemisia tabaci is a worldwide serious pest possessing more than 24 biotypes; these biotypes are difficult to distinguish by morphological characters only. In this study, we designed the specific primer sets for each of 6 B. tabaci biotypes (A, B, Q, Na, AN, S). Two primer sets, BaAF/BaAR and BaQF/BaQR, for A and Q biotypes, respectively, were based on the sequences of RAPD specific fragments. Four forward primers, BaBF, BaNaF, BaANF, and BaSF, with a common reverse primer, L2-N-3014, for other 4 biotypes, B, Nauru, AN, and S, respectively, were based on their own COI sequence. The 16S ribosomal DNA (rDNA) of the secondary endosymbiont and Wolbachia-specific gene, wsp (cell surface protein of Wolbachia), were amplified and sequenced. Based on the occurrences of 16S rDNA and wsp by PCR, more than 55% and 38% of examined B. tabaci (202 populations) harbored secondary endosymbiont and Wolbachia, respectively. Molecular phylogenetic analysis of 16S rDNA sequences showed that the secondary endosymbiont was located in the clade of γ-3 subdivision of the Proteobacteria. wsp gene sequences, revealed that the Wolbachia found in B. tabaci populations of Taiwan belongs to the group B and subgroup Con/Rug. The results also showed that the carrying/infection rate of secondary endosymbiont/Wolbachia might be related to the biotype of B. tabaci. | en |
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| dc.description.tableofcontents | 目錄
中文摘要 英文摘要 目錄 ------------------------------------------------------------------------------------------- i 表次 ----------------------------------------------------------------------------------------- iv 圖次 ------------------------------------------------------------------------------------------ v 壹、前言 ------------------------------------------------------------------------------------ 1 貳、往昔研究 ------------------------------------------------------------------------------ 4 一、煙草粉蝨種群 --------------------------------------------------------------------- 4 二、煙草粉蝨生物小種之鑑定 ------------------------------------------------------ 5 三、專一性引子組 --------------------------------------------------------------------- 8 四、內共生菌 (Endosymbionts) ---------------------------------------------------- 8 (一) 初級內共生菌 (Primary endosymionts) --------------------------------- 9 (二) 生殖性寄生 (Reproductive parasitism) ---------------------------------- 10 (三) 二級內共生菌 (Secondary endosymbionts) ----------------------------- 11 參、材料與方法 --------------------------------------------------------------------------- 13 一、粉蝨標本之採集與保存 --------------------------------------------------------- 13 二、粉蝨基因組 DNA 之萃取 ------------------------------------------------------ 14 三、膠體的製備與電泳 --------------------------------------------------------------- 14 四、專一性引子組 -------------------------------------------------------------------- 15 (一) DNA 逢機增幅多態型聚合酶連鎖反應 (RAPD-PCR) 分析 ------- 15 (二) PCR 產物之純化 ------------------------------------------------------------ 15 (三) 接合反應 (Ligation) -------------------------------------------------------- 16 (四) 質體轉形 (Transformation) ------------------------------------------------ 16 (五) 重組質體鑑定 ---------------------------------------------------------------- 16 (六) DNA 定序及引子組之設計 ------------------------------------------------ 17 (七) 專一性引子組之測試 ------------------------------------------------------- 17 五、煙草粉蝨內共生菌 (Endosymbionts) 之偵測 ------------------------------ 18 (一) 二級共生菌 (Secondary endosymbionts) ----------------------------------- 18 (二) Wolbachia ------------------------------------------------------------------------- 19 (三) 類緣關係樹 (Phylogenetic tree) 之重建 ----------------------------------- 20 肆、結果 ----------------------------------------------------------------------------------- 21 一、煙草粉蝨標本之採集 ----------------------------------------------------------- 21 二、專一性引子組之開發 ------------------------------------------------------------ 21 (一) RAPD-PCR 分析 ------------------------------------------------------------ 21 (二) 序列分析 ---------------------------------------------------------------------- 23 (三) 專一性引子組之設計 ------------------------------------------------------- 23 (四) 專一性引子組之測試 ------------------------------------------------------- 24 三、臺灣地區煙草粉蝨之內共生菌 ----------------------------------------------- 39 (一) 二級內共生菌 (Secondary endosymbionts) ----------------------------- 39 (二) Wolbachia ---------------------------------------------------------------------- 40 伍、討論 ----------------------------------------------------------------------------------- 46 一、臺灣地區煙草粉蝨之分布與寄主範圍 -------------------------------------- 46 二、專一性引子組之開發 ----------------------------------------------------------- 47 三、臺灣地區煙草粉蝨之內共生菌 ------------------------------------------------ 49 (一) 二級內共生菌 (Secondary endosymbionts) ----------------------------- 49 (二) Wolbachia ---------------------------------------------------------------------- 51 陸、結論 ----------------------------------------------------------------------------------- 54 柒、參考文獻 ----------------------------------------------------------------------------- 55 捌、誌謝 ------------------------------------------------------------------------------------ 65 玖、附錄 ----------------------------------------------------------------------------------- 66 附錄一、以 OPA15 增幅之煙草粉蝨 A 生物小種特有 DNA 片段序列 --------------------------------------------------------------------------------------- 66 附錄二、以 OPO18 增幅之煙草粉蝨 Q 生物小種特有 DNA 片段序列 --------------------------------------------------------------------------------------- 67 附錄三、煙草粉蝨 B、Nauru、AN 及 S 生物小種粒線體 COI 序列 ------ 68 附錄四、臺灣地區煙草粉蝨二級內共生菌 16S rDNA 片段序列比對之結果 --------------------------------------------------------------------------------------- 70 附錄五、臺灣地區煙草粉蝨內共生菌 Wolbachia 之 wsp 基因序列比對之結果 --------------------------------------------------------------------------------------- 72 附錄六、臺灣地區煙草粉蝨族群受內共生菌感染之比例 --------------------- 73 附錄七、本實驗分析之臺灣地區煙草粉蝨族群 --------------------------------- 74 表次 表一、本實驗分析之國外煙草粉蝨族群 --------------------------------------------- 13 表二、本實驗分析之臺灣地區粉蝨種類 --------------------------------------------- 14 表三、研究中所使用區分不同煙草粉蝨生物小種之引子序列及其黏合條件 ------------------------------------------------------------------------------------------ 18 表四、本研究中偵測煙草粉蝨二級內共生菌與 Wolbachia 所使用之引子序列及其黏合條件 ------------------------------------------------------------------------ 19 圖次 圖一、臺灣地區煙草粉蝨之採樣分布圖 --------------------------------------------- 22 圖二、以逢機引子 OPA15 增幅煙草粉蝨之 RAPD 電泳圖譜 ---------------- 25 圖三、以逢機引子 OPO18 增幅煙草粉蝨之 RAPD 電泳圖譜 ---------------- 26 圖四、以引子組 BaAF/BaAR 區分不同煙草粉蝨生物小種 -------------------- 27 圖五、以引子組 BaQF/BaQR 區分不同煙草粉蝨生物小種 -------------------- 28 圖六、以引子組 BaBF/L2-N-3014 區分不同煙草粉蝨生物小種 --------------- 29 圖七、以引子組 BaNaF/L2-N-3014 區分不同煙草粉蝨生物小種 ------------- 30 圖八、以引子組 BaANF/L2-N-3014 區分不同煙草粉蝨生物小種 ------------ 31 圖九、以引子組 BaSF/L2-N-3014 區分不同煙草粉蝨生物小種 --------------- 32 圖十、以引子組 BaAF/BaAR 區分不同粉蝨種類 -------------------------------- 33 圖十一、以引子組 BaQF/BaQR 區分不同種粉蝨類 ----------------------------- 34 圖十二、以引子組 BaBF/L2-N-3014 區分不同粉蝨種類 ------------------------ 35 圖十三、以引子組 BaNaF/L2-N-3014 區分不同粉蝨種類 ---------------------- 36 圖十四、以引子組 BaANF/L2-N-3014 區分不同粉蝨種類 --------------------- 37 圖十五、以引子組 BaSF/L2-N-3014 區分不同粉蝨種類 ------------------------ 38 圖十六、煙草粉蝨二級內共生菌之 16S rDNA 及 Wolbachia 之偵測電泳圖 ------------------------------------------------------------------------------------------ 43 圖十七、利用煙草粉蝨二級內共生菌之 16S rDNA 序列以 Neighbor-joining 法重建之親緣關係樹 --------------------------------------------------------------- 44 圖十八、利用利用 wsp 基因序列以 Neighbor-joining 法重建之親緣關係樹 ------------------------------------------------------------------------------------------ 45 | |
| dc.language.iso | zh-TW | |
| dc.title | 煙草粉蝨生物小種之分子鑑定及其二級內共生菌之初探 | zh_TW |
| dc.title | Studies on the molecular identification and secondary endosymbionts of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 93-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 王重雄(Chung-Hsiung Wang) | |
| dc.contributor.oralexamcommittee | 張慧羽,路光暉,賴吉永,葉文斌 | |
| dc.subject.keyword | 煙草粉蝨,生物小種,分子鑑定,內共生菌, | zh_TW |
| dc.subject.keyword | Bemisia tabaci,biotype,molecular identification,endosymbiont,Wolbachia, | en |
| dc.relation.page | 84 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2005-07-14 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 昆蟲學研究所 | zh_TW |
| 顯示於系所單位： | 昆蟲學系 | |
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