臺灣博碩士論文加值系統

擬細鯽(Nicholsicypris normalis)為中國原生淡水魚類，多分布於海南島及中國華南地區，由於此物種對於水質的適應性良好、產卵數多、生存性高，其族群的個體數在河流中相較於其他物種具有相對優勢，在自然平衡中占有重要的地位，而對於此物種在基因多樣性與族群遺傳上的相關研究卻甚少。本研究採用粒線體基因組多個基因區塊針對中國各地區擬細鯽族群進行分子親緣研究，為探討可能之親緣地理模式，粒線體基因選取控制區域及細胞色素b基因，總共分別從9個不同地點採集107個樣本，並鑑定出105個mtDNA單倍型，透過聚類分析(clustering algorithm)與F-統計(fixation indices)的族群分化指數NST(0.532)及GST(0.004)可反映出的擬細鯽系統發育與地理學有顯著的關係，利用S-DIVA顯示擬細鯽的祖先支系可能源自於海南島與雷州半島以東，並使用近似貝葉斯計算法重建擬細鯽族群的演化歷史，結果顯示(1)擬細鯽之祖先分布於海南島北部；(2)海南島的五指山與鶯歌嶺山脈是限制物種基因交流的重要屏障；(3)在瓊州海峽地面下降之前，雷州半島、瓊州海峽和海南島長期連接形成一大塊陸地；(4)在冰河時期，大陸棚暴露造成擬細鯽族群之分散。
關鍵字：擬細鯽、親緣地理、粒線體基因、近似貝葉斯分析

Nicholsicypris normalis (Teleostei: Cyprinidae) is an original freshwater fish in Hainan Island and southern China. N. normalis have more advantageous than other freshwater fish because of the great adaptation to the river, high fertility rate and survival rate let own a high quantity. N. normalis have an important status of balance of natural, but little has been known about its genetic structure. The molecular genetic study is based on multigenic technology for exploring about the phylogeogrphy of N. normalis in the different area in China. This study examined the phylogeography of N. normalis using sequences of the mitochondrial DNA control region and cytochrome b gene. In total, 107 specimens were collected from nine populations. All 105 mtDNA haplotypes were identified. The phylogenetic trees reconstructed with different clustering methods were not identical. This result revealed an admixture-like genetic structure, but a comparison of the fixation indices NST (0.532) and GST (0.004) revealed that the phylogeny and geography had a significant relationship. The results of a statistical dispersal-vicariance analysis suggested that the ancestral populations of N. normalis were distributed widely in Hainan Island and east of Leizhou Peninsular. The approximate Bayesian computation approaches were used to reconstruct the history of N. normalis. Our study (1) found that the ancestral populations were distributed northern Hainan Island; (2) showed that the Wuzhishan and Yinggeling Mountains Range was an important barrier limiting gene exchanges among populations; (3) implied that before the ground of Qiongzhou Strait fell, the Leizhou Peninsula, Qiongzhou Stait, and Hainan Island connected as Hainan Peninsula for a long time; and (4) indicated that during glaciation, the continental shelves were exposed, which contributed with the dispersion of populations.

Key words: Nicholsicypris normalis, phylogeogrphy, mitochondrial DNA, Approximate Bayesian computation

目錄
致謝 i
摘要 iii
Abstract iv
目錄 vi
表目錄 viii
圖目錄 ix
第一章　前言 1
1.1　親緣地理學 1
1.1.1　遺傳標誌 2
1.1.2　聚合酶連鎖反應技術 3
1.2　鯉科魚類 6
1.2.1　擬細鯽 6
1.3　地理環境 7
1.3.1　海南島 8
1.4　研究目的 10
第二章　實驗方法與設計 11
2.1　樣品採集 11
2.2　樣品萃取 13
2.3　聚合酶鏈鎖反應 15
2.4　純化 17
2.5　基因分析 18
第三章　結果與討論 22
3.1　序列分析 22
3.2　單倍型歧異度與核苷酸歧異度 22
3.3　遺傳分化指數 25
3.4　最大似然法與貝葉斯分析 26
3.5　溯祖時間 30
3.6　分子變異分析結果 31
3.7　近似貝葉斯分析結果 33
第四章　結論 35
第五章　參考文獻 37

 
表目錄
表一、樣品數量及採集點表 12
表二、實驗之引子 15
表三、PCR反應設定 16
表四、擬細鯽單倍型歧異度與核苷酸歧異度結果 23
表五、族群之間之FST值表 25
表六、分子變異分析(AMOVA) 32
表七、近似貝葉斯計算結果 34


 
圖目錄
圖一、聚合酶連鎖反應示意圖 5
圖二、擬細鯽採樣地點 7
圖三、擬細鯽單倍群分布圖 23
圖四、貝葉斯計算演化樹結果 27
圖五、最大似然法計算演化樹結果 28
圖六、單倍型網絡 29
圖七、擬細鯽族群之演化路徑 30

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