台灣黑眶蟾蜍之親緣地理研究 魏勢璋、賴伯琦
E-mail: [email protected]
摘 要
黑眶蟾蜍 (Duttaphrynus melanostictus) 現今廣泛分布於中國南部及東南亞， 在台灣主要分布於五百公尺以下的闊葉林或開 墾地。 本研究於台灣採集到的地點分別來自不同縣市的 23 個族群，並利用粒線體 DNA 中 cytochrome b 和 control region 之間片段，長度為 982 bp。由 90 個定序樣本 所得的序列發現有 23 個變異位置 (variable sites)，18 種單倍型 (haplotypes) ， 其中僅單倍型 Hp01 廣泛分布於台灣以及蘭嶼、綠島，並佔了所有序列的 46.7%。 透過 Bayesian skyline plot 顯示台灣黑眶 蟾蜍在末次冰期後族群量顯著地增加， 推測是因氣候穩定所致。而基因多樣性與族群變動分析則顯示台灣僅有西部族群發 生過族群擴張現象，推測是由少數之族群快速發展而來。經計算 Kimura-2-parameter 基因距離後再以 neighbor-joining 及 maximum-parsimony 方法重建單倍型間之樹狀 圖後發現所取樣的黑眶蟾蜍序列與採集地點間沒有明顯的親緣分化現象，但 在每個地區或鄰近地區 (除了雲林以外) 都發現當地特有之單倍型。經由分析親緣網狀圖推測黑眶蟾蜍進入台灣地區可能 有 兩條播遷路徑： 1. 由台灣中部地區進入，並向東散播，隨後才向南拓殖，最後才往北擴散 2. 從台灣南部地區進入，並先 後向北與向南散播，其次才向東擴散，最後又由宜蘭播遷回大陸。 由於目前所得之研究資料僅得福建地區部分族群，若欲 進一步推斷台灣之黑眶蟾蜍應是否至少 有兩次遷入現象，尚須大陸其他沿海地區樣本以進行更深入的探討。
關鍵詞 : 黑眶蟾蜍、親緣地理、中心散播假說、同宗效應假說、粒線體 DNA 分析 目錄
授權書...iii 中文摘要...iv Abstract...vi 誌謝...vii 目 錄...viii 圖目錄...xi 表目錄...xii 1. 前言...1 1.1 地理分 布...1 1.2 親緣歷史與分類探究...1 1.3 台灣地質歷史與物種播遷...2 1.4 古地理事件與生物地理...4 1.5 分子演化與粒線體 DNA...6 1.6 親緣地理學...8 1.7 研究目的...10 2. 實驗方法...11 2.1 樣本收集...11 2.2 粒線體 DNA 之萃取...11 2.3 聚合酵素連鎖反應 (Polymerase Chain Reaction)....12 2.3.1 引子設計...12 2.3.2 PCR 條件之設定...12 2.3.3 配製 1% 瓊脂凝膠...13 2.3.4 PCR 產物電泳分 析...13 2.4 DNA 定序...13 2.5 分子資料分析...13 2.5.1 序列校正...13 2.5.2 親緣樹之建 立...14 2.5.3 親緣網狀圖之建立...14 2.5.4 多樣性 (diversity) 統計分析....15 2.5.5 中性測試...15 2.5.6 分歧時間之估計...16 3. 結果...18 3.1 序列變異...18 3.2 親緣關係...18 3.3 族 群之遺傳變異...20 3.4 分子變方分析 (AMOVA)...21 3.5 族群分化...21 4. 討論...23 4.1 台灣黑眶蟾蜍之親緣地理...23 4.2 族群遺傳結構...25 4.3 族群分化之探討...29 4.4 島內族群散播歷 史...32 5. 結論...35 參考文獻...59 附錄...70
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