臺灣博碩士論文加值系統

為了瞭解澳洲東部的三種淡水鰻 (Anguilla spp)：熱帶性澳洲花鰻 (A. reinhardtii, Steindachner 1867)，以及溫帶性澳洲短鰭鰻 (A. australis, Richardson 1841) 和紐西蘭大鰻 (A. dieffenbachii, Gray 1842) 的族群遺傳結構及其演化關係，本研究以微衛星DNA為遺傳標記，分析6-7個微衛星基因座的遺傳變異性。總共使用了1277尾鰻線標本，分別是1997-1999年採自澳洲六個河口的澳洲花鰻 (799尾) 和澳洲短鰭鰻 (287尾) 以及1995-1996年採自紐西蘭南北島的三個河口澳洲短鰭鰻 (101尾) 和紐西蘭大鰻 (90尾)。其中，澳洲東岸Albert River的澳洲花鰻標本連續採集12個月以上。

分析結果發現澳洲花鰻雖然終年產卵，但其族群遺傳結構並無時間或空間的分化現象，這可能和其仔魚浮游期短、散佈範圍窄以及終年產卵基因容易交流有關。相反地，澳洲短鰭鰻的仔魚漂浮期長、分佈域含蓋澳洲及紐西蘭兩地，地區間卻有明顯的族群遺傳分化現象。紐西蘭大鰻為已知仔魚浮游期最長的淡水鰻，但族群分化情形並不明顯，可能和可散佈陸域範圍太小 (僅局限於紐西蘭島) 有關。紐西蘭大鰻的外部型態及其基因結構比同一個演化支序的澳洲短鰭鰻保留更多的祖徵。顯示溫帶性紐西蘭大鰻可能是上述鰻鱺屬 (Anguilla) 的三個種類中最早抵達紐澳地區的的淡水鰻。本研究結果顯示微衛星基因座的等位基因變異特性，可以提供澳洲三種淡水鰻的族群擴散及演化的部分訊息。

Population genetic structure and evolutionary scenario were investigated with the microsatellites analysis for three species of the freshwater eels, tropical Anguilla reinhardtii (Steindachner 1867), and temperate Anguilla australis (Richardson 1841) and Anguilla dieffenbachii (Gray 1842) in Australia and New Zealand. A total of 1277 glass eel specimens were used: 388 from East Australia and New Zealand for A. australis, 90 from both North and South Island of New Zealand for A. dieffenbachii, and 799 from East Australia for A. reinhardtii, including a series of monthly collections for over 12 months from the Albert River estuary. The specimens were collected from East Australis in 1997-1999 and New Zealand in 1995-1996.
For A. reinhardtii there was no significant spatial and temporal differentiation in the genetic structure. This may be due to the fact that it has a short pelagic larval duration, narrow distribution range, but a long, all the year round spawning period, that facilitate the gene flow. For A. australis that has comparatively longer pelagic larval duration than that of A. reinhardtii, two geographical populations were recognized in East Australia and New Zealand, respectively. For A. dieffenbachii in New Zealand that has the longest duration of the pelagic larval stage has no population genetic differentiation due to the small land mass for dispersal. A. dieffenbachii is characterized with more ancient morphological and genetic characters than the A. australis in the same evolutionary lineage. These may suggest that temperate eel A. dieffenbachii is the earliest arrival of the freshwater eel in Oceania. Genetic variation in microsatellite loci may provide a useful tool in examining evolution and population dispersal of freshwater eels.

口試委員會審定書………………………………………………………………………i
誌謝……………………………………………………………………………………...ii
中文摘要………………………………………………………………………………..iii
英文摘要……………………………………………………………………..…….…...iv
一、前言........……………………………………………………………………...……1
1. 淡水鰻的類緣關係及其生活史的比較………………….………….……...…1
2. 淡水鰻族群遺傳研究的進展…………………………..………….……….….3
3. 澳洲東部三種淡水鰻的分佈及生活史特徵……………………………….…5
4. 問題與假設……………….……………………………………………………7
5. 研究動機與目的…………………………………………...…….…………….9
二、材料與方法…………………………………………………………………….….11
1. 鰻線之採集與保存……………………………………………………...…....11
2. 微衛星DNA的特性………………………….………………….….…….….11
3. 微衛星DNA基因座上基因型之分析……….………………….….…….…..13
3.1 DNA萃取………………………………….……………….….…………13
3.2 聚合酶連鎖反應 (Polymerase Chain Reaction, PCR) 及基因型分析 (genotyping)…………………………………………………………….13
3.3 微衛星DNA基因片段的定序……………………………………….…14
4. 資料分析…………………………………………………………….………..14
4.1 遺傳變異性 （genetic variability） 的分析…………………………......14
4.2 哈溫平衡（Hardy-Weinberg equilibrium）的檢定…………….………15
4.3 族群間的分化程度……………………………………………………...16
4.4 遺傳距離 (genetic distance) 的計算與親緣關係樹 (phylogenetic tree) 的建構………………………………………………………………….16
4.5 連結不平衡 (linkage disequilibrium) 測試……….………….……..…17
4.6 無效等位基因及評分錯誤 (null allele or scoring error) 之偵測……...17
4.7 遺傳距離與時空距離關係之測試……………………………………...17
4.8 近交之檢定…………………………………………………………...…17
4.9 來自不同族群的個體之分配測試 (assignment test)…………………..18
三、結果…………………………………………………………………………….…19
1. 澳洲花鰻的族群遺傳結構……………………………………………….......19
1.1 遺傳變異性及哈溫平衡檢定……………………………………...........19
1.2 族群遺傳結搆的時空變化………………………………………….…..20
1.3 樣本間的遺傳距離..………………………………………………….....20
1.4 時間樣本間的遺傳距離…………………………………………….…..21
1.5 地點之間的遺傳距離……………………………………………….......21
2. 澳洲短鰭鰻的族群遺傳結構…………………………………………….......22
2.1 遺傳變異性及哈溫平衡檢定…………………………………………...22
2.2 族群遺傳分化現象………………………………………………...........23
2.3 成對遺傳距離和成對遺傳分化指數的顯著性差異…………………...23
2.4 澳洲短鰭鰻的親緣地理………...…………………………………..…..24
3. 大洋洲兩種溫帶性澳洲短鰭鰻及紐西蘭大鰻族群遺傳變異特性的比較……………………………………………………………………………..24
3.1 參數變異的種間比較……………………………………………….......25
3.2 等位基因種類及頻率的種間比較………………………………….......26
3.3 微衛星基因座序列的種間比較…………………………………….......26
3.4 澳洲短鰭鰻和紐西蘭大鰻的族群遺傳結構的比較……………….......26
四、討論……………………………………………………………………………….28
1. 淡水鰻族群的特性…………………………………………………………...28
2. 澳洲花鰻族群遺傳結構之探討…………………………...............................29
2.1 澳洲花鰻微衛星基因座等位基因多型性及序列變化…………………………………………………...…………………..29
2.2 微衛星基因座數及樣本數在澳洲花鰻族群遺傳結構分析之探討……………………………………………………………………….30
2.3 澳洲花鰻族群遺傳結構不具時空變化的可能原因…………..…..…...30
3. 澳洲短鰭鰻族群結構之探討………………………………………………...31
3.1 時間效應是否對族群遺傳結構產生影響？……….………………......32
3.2 族群遺傳分化指數的比較………………………….………………......32
3.3 仔魚從外洋產卵場至河口域的可能漂送途徑…….………………......33
4. 澳洲短鰭鰻和紐西蘭大鰻的演化秩序……………………….……………..35
4.1 大西洋及大洋洲兩個鰻魚支序的演化歷史之比較…….………...…...35
4.2 大洋洲的澳洲短鰭鰻由紐西蘭大鰻演化而來的可能性..………..…...36
4.3 澳洲短鰭鰻族群膨脹的證據……………………………………...…....36
4.4 族群分化程度和陸地效應的關係…………………………………...…37
5. 三種澳洲鰻可能的種化路線與地質年代的關係………………………...…38
五、總結論…………………………………………………………………………….40
六、參考資料………………………………………………………………………….41
七、表………………………………………………………………………………….56
八、圖………………………………………………………………………………….77
九、附錄……………………………………………………………………………97

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