臺灣長鬃山羊（Capricornis swinhoei）圈養族群於非侵入式遺傳分析暨微衛星標幟開發之探討

Study on Non-invasive Genetic Analysis and Microsatellite Marker Development in Captive Population of Formosan Serow (Capricornis swinhoei)

10.6342/NTU.2011.01550

張怡穎

臺灣長鬃山羊 ； 圈養族群 ； 非侵入式遺傳分析 ； 微衛星標幟 ； 族群結構 ； Formosan serow ； captive population ； non-invasive genetic study ； microsatellite markers ； genetic structure

國立臺灣大學動物科學技術學系學位論文

2011年

碩士

王佩華

繁體中文

臺灣長鬃山羊為臺灣特有種，在國內屬珍貴稀有保育動物。為維持其族群之遺傳多態性，及未來可能進行域外保育，了解圈養（臺北市立動物園）與野外臺灣長鬃山羊的遺傳現況並避免發生近親繁殖便成一重要課題。然而臺灣長鬃山羊為生性敏感之物種，不易進行捕捉採樣；此外，臺灣長鬃山羊目前仍欠缺進行個體層級遺傳分析之分子標幟，而目前分子標幟中以微衛星標幟較為廣泛使用。因此本試驗之目的便為建立自臺灣長鬃山羊糞樣萃取 DNA 之非侵入式分析方法並發展適用於臺灣長鬃山羊個體及族群遺傳分析之微衛星標幟。 本試驗共評估三種保存方法與兩種自糞便萃取 DNA 之方法，並進行長達四週之保存試驗。試驗結果顯示保存、萃取方法與保存時間皆顯著影響 PCR 效果（P < 0.0001）。在本次試驗所測試方法中，糞樣以 95% 乙醇保存，並以 2CTAB/PCI 法進行萃取，至少於四週內仍可得最佳 PCR 結果。此法成本低廉且易於操作，除了應用於園內遺傳監測之採樣外，亦可作為未來野外非侵入式遺傳分析之先期研究。 微衛星標幟測試部分，利用來自韓國長鬃山羊與跨物種微衛星標幟雖可初步了解臺北市立動物園內之臺灣長鬃山羊遺傳結構，但可能由於多態性不足（平均 PIC = 0.562），加上嚴重連鎖不平衡情況，無法進一步完成個體鑑別與親子鑑別。 而本研究自行開發之 23 個交替基因個數大於 3 之微衛星標幟，經檢測臺北市立動物園 43 頭臺灣長鬃山羊，共有 21 組高多態性微衛星標幟。且於個體鑑別與親子鑑別時，亦優於跨物種微衛星標幟。 整合 6 組跨物種微衛星標幟與 17 組自行開發之微衛星標幟所得之 23 組微衛星標幟進行遺傳分析，顯示目前臺北市立動物園內臺灣長鬃山羊族群尚無嚴重近親配種情況，但族群已產生些微分化，反映近年來特定配對之配種。此外，個體鑑別部份，合併之 23 組微衛星標幟可用於建立臺北市立動物園內臺灣長鬃山羊個體之分子身份識別；親子鑑別部份，在已知母畜之情況下，親緣鑑別之結果與現有系譜幾乎可相互印證（93.5%）。 綜合以上所述，合併後的 23 組微衛星標幟在臺灣長鬃山羊圈養族群的動物管理上可輔佐動物園現有晶片系統，作為動物樣本端或新生仔畜分子身份辨識工具，或用於推測系譜、分析園內個體間之遺傳關係，做為園內個體配種參考。配合本試驗所建立之非侵入式遺傳分析方法，日後或許亦可使用此些微衛星標幟於野外臺灣長鬃山羊族群分析，並規劃野放或野外引種等措施，以同時維持圈養與野外族群之遺傳多態性，達成域外保育之目的。

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