柯俊成臺灣大學：昆蟲學研究所謝佳宏Hsien, Chia-HungChia-HungHsien2007-11-262018-06-292007-11-262018-06-292004http://ntur.lib.ntu.edu.tw//handle/246246/55050煙草粉蝨 (Bemisia tabaci) 為泛世界分佈的昆蟲，鑑定特徵主要是依據第四齡若蟲之形態，但外部形態容易因寄主植物之不同而產生差異，造成鑑定上之困難。因此，近年來隨著分子生物學與生物技術方法之發展，分子標示亦被用來研究煙草粉蝨。本研究全面採集臺灣地區煙草粉蝨，利用分子標示來進行親緣關係、族群遺傳及親緣地理學之研究。經調查發現煙草粉蝨於臺灣地區廣泛分布。利用 3 種分子標示所作之親緣關係樹分析結果，粒線體 COI、16S rDNA 與核糖體 ITS1 皆可區分煙草粉蝨生物小種 (biotypes)，並指出臺灣地區目前有3生物小種，分別為 B、AN 及Nauru 生物小種。遺傳距離分析結果，B 生物小種內遺傳差異小， Nauru 與 AN 生物小種相對差異較大，推測 B 生物小種為近期外來侵入，而 AN 與 Nauru 生物小種則為早期自然擴散侵入。分子變方分析與族群分化指數，顯示生物小種間遺傳差異顯著。另一方面，Nauru 生物小種族群遺傳研究顯示，臺灣西部族群基因交流順暢，而西部與東部遺傳上差異大，基因交流情形符合隔離分化模式，地形與地理距離會影響族群基因交流，但東部與馬祖族群遺傳差異小，則不符合隔離分化模式。親緣地理學之探討，Nauru 生物小種存在多次入侵事件，除了自然擴散以外，亦可能有人為擴散因素，未來仍需更多單基因型與更大範圍地理區之採集調查才能更進一步證實。Bemisia tabaci is global distribution, the identification of character is based on the morphology of pupal case. However, this species is difficult to identify because the morphological characters could be polymorphic in different host plants. Recently, molecular markers are developed to study this species. The aim of this research was to study phylogeny, population genetics, and phylogeography by molecular markers. B. tabaci was collected from many regions in Taiwan, and three kinds of molecular markers were selected to analysis. Phylogenetic analyses of mitochondrial COI, 16S rDNA, and ribosomal ITS1 reveal three biotypes, B, AN, and Nauru biotypes in Taiwan. Genetic distance analysis reveals the relationship within populations of B biotype is closely related. On the contrary, genetic distance within populations of AN, and Nauru biotypes is more variable. The result could be speculated that B biotype moved into Taiwan recently. On the other hand, AN, and Nauru biotypes colonized into Taiwan naturally. AMOVA and FST analyses reveal that genetic variance between biotypes is significant. The population genetic studies of Nauru biotype reveal that gene flow is smooth between populations in the western Taiwan, and limited between the western, and eastern Taiwan. The result confirms to the theory of isolation by distance. Geographical region, and distance will limit the gene flow between populations, but gene flow between Matsu, and the eastern Taiwan populations don’t answer to the theory. Phylogeography analyses reveal that Nauru biotype colonized into Taiwan many times. The Nauru biotype colonized into Taiwan either by nature or international trade. However, phylogeographical studies of Nauru biotype still need more samples to be analyzed in the future.中文摘要 英文摘要 目錄 ----------------------------------------------------------------------------------- I 表次 ----------------------------------------------------------------------------------- III 圖次 ----------------------------------------------------------------------------------- IV 一、前言 ------------------------------------------------------------------------------ 1 二、往昔研究 ------------------------------------------------------------------------ 4 一、歷史回顧 ------------------------------------------------------------------ 4 二、煙草粉蝨種群 ----------------------------------------------------------- 5 三、寄主植物 ----------------------------------------------------------------- 5 四、傳播之植物病毒 -------------------------------------------------------- 6 五、內共生物 ------------------------------------------------------------------ 7 六、分子標示在煙草粉蝨之研究 ---------------------------------------- 7 七、煙草粉蝨之起源 -------------------------------------------------------- 10 三、材料與方法 --------------------------------------------------------------------- 12 一、標本採集及保存 --------------------------------------------------------- 12 二、實驗方法 ------------------------------------------------------------------ 12 三、資料分析 ------------------------------------------------------------------ 14 四、結果 ------------------------------------------------------------------------------ 20 一、臺灣地區煙草粉蝨分布情形 ----------------------------------------- 20 二、序列分析 ---------------------------------------------------------------- 20 三、親緣關係之研究 -------------------------------------------------------- 27 四、族群遺傳之研究 ------------------------------------------------------- 33 五、親緣地理學之研究 ----------------------------------------------------- 43 五、討論 ----------------------------------------------------------------------------- 48 一、臺灣地區煙草粉蝨分布情形 ----------------------------------------- 48 二、序列比較 ----------------------------------------------------------------- 48 三、親緣關係之研究 ------------------------------------------------------- 49 四、族群遺傳之研究 ------------------------------------------------------ 51 五、分子變方分析 --------------------------------------------------------- 53 六、親緣地理學之探討 --------------------------------------------------- 54 六、結論 ----------------------------------------------------------------------------- 57 七、參考文獻 ----------------------------------------------------------------------- 58 八、致謝 ---------------------------------------------------------------------------- 70 九、附錄 ----------------------------------------------------------------------------- 71 附錄一、臺灣地區煙草粉蝨 Nauru 生物小種粒線體 COI 序列 之19 種單倍型 --------------------------------------------------- 71 附錄二、臺灣地區煙草粉蝨之採集紀錄 -------------------------------- 74 附錄三、國外煙草粉蝨之採集紀錄 -------------------------------------- 80 附錄四、本研究所使用之煙草粉蝨 Nauru 生物小種各族群單倍 型之分布 ------------------------------------------------------------ 81 附錄五、GenBank下載之煙草粉蝨粒線體 COI 序列 ----------------- 84 附錄六、GenBank下載之煙草粉蝨rDNA ITS1 序列 ----------------- 85 附錄七、GenBank下載之煙草粉蝨粒線體 16S 序列 ----------------- 871911909 bytesapplication/pdfen-US親緣地理學族群遺傳親緣關係分子標示煙草粉蝨phylogeographypopulation geneticsphylogenymolecular markerBemisia tabacithesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/55050/1/ntu-93-R91632005-1.pdf