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研究生: 簡竹君
Chien, Chu-Chun
論文名稱: 微衛星DNA選殖與台灣二倍體傅氏鳳尾蕨族群遺傳結構
Characterization of microsatellite DNA and genetic structure of diploid Pteris faurieri in Taiwan
指導教授: 蔣鎮宇
Chiang, Tzen-Yuh
學位類別: 碩士
Master
系所名稱: 生物科學與科技學院 - 生物多樣性研究所
Institute of Biodiversity
論文出版年: 2010
畢業學年度: 98
語文別: 中文
論文頁數: 44
中文關鍵詞: 跨種擴增微衛星三倍體傅氏鳳尾蕨二倍體傅氏鳳尾蕨族群遺傳結構台灣
外文關鍵詞: Cross-ampliciation, microsatellites, Pteris fauriei var. Faurie, Pteris fauriei var. minor, population genetic structure, Taiwan
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  • 台灣地區傅氏鳳尾蕨 (Pteris fauriei)有兩個變種,分別minor和fauriei。P. fauriei var. minor為二倍體,P. fauriei var. fauriei則為三倍體。二倍體植株常分布於較溫暖的地區,三倍體植株則生長於草地和林下。二倍體傅氏鳳尾蕨在台灣本島與鄰近島嶼呈不連續分布,不分布於台灣中部和馬祖地區。三倍體傅氏鳳尾蕨則沒有分布於台灣南端和東部島嶼。本研究著重於二倍體傅氏鳳尾蕨的族群遺傳研究並討論三倍體的起源。由P. fauriei var. minor篩選出11組具多型性的微衛星基因座應用於族群遺傳分析。其中有4組基因座在21個鳳尾蕨屬物種中能被擴增出來,且所有基因座皆能在三倍體傅氏鳳尾蕨中被擴增。於台灣地區二倍體傅氏鳳尾蕨遺傳多樣性分析中,基因座平均多樣性 (average gene diversity over loci)偏高 (0.772),基因豐富度 (allelic richness)介於4.071到12.221和稀有對偶基因數多 (rare allele, 1到9個)。AMOVA的結果顯示遺傳變異主要累積在族群內 (91.75%),使得族群間有低的族群平均分化指數(FST= 0.083)。而STRUCTURE分析結果中,沒有地理分化亦支持此結果。 總結而言,推測三倍體傅氏鳳尾蕨應為二倍體傅氏鳳尾蕨自體多倍化而形成。低的族群分化指數,顯示P. fauriei var. minor台灣族群間有暢通的基因交流。

    There are two varieties of Pteris fauriei in Taiwan, i.e. fauriei and minor. P. fauriei var. minor is diploid and P. fauriei var. faurie is triploid. In general, diploid plants occur in warmer habitats, while triploid plants grow at grassland and woodland. P. fauriei var. minor is widespread in Taiwan and nearby islets, except for central Taiwan and Matsu islands. In contrast, P. fauriei var. fauri is not found in southern Taiwan or islets in the Taiwan Strait. This study focuses on the population genetics of P. fauriei var. minor and the origin of P. fauriei var. faurie. In this study, 11 microsatellite loci were isolated from P. fauriei var. minor for population genetic analysis and can also be amplified with P. fauriei var. faurie. Four were cross-amplified with 21 Pteris species. The average of gene diversity is high (0.772), the number of allelic richness from 4.071 to 12.221, and rare allele number is also high (1 to 9). AMOVA indicates that most genetic variation resides within populations (91.75%), resulting in low genetic differentiation (FST= 0.083). Lack of geographical subdivision was further supported by STRUCTURE analysis. In conclusion, all loci can be amplified with P. fauriei var. fauriei, suggesting its autopolyploidy. Low population differentiation is likely due to frequent gene flow between P. fauriei var. minor populations.

    目 錄 中文摘要 I 英文摘要 II 誌謝 III 目錄 IV 表目錄 VII 圖目錄 VIII 壹、前言 1 一、族群遺傳學 1 二、分子標記的應用 1 三、台灣地區傅氏鳳尾蕨之介紹 4 四、蕨類的無性種 6 五、研究目的 7 貳、材料與方法 8 一、研究材料 8 二、研究方法 8 2.1植物總DNA之萃取 8 2.2微衛星體基因座之篩選 9 2.2.1 DNA的擴增(聚合酶鏈連鎖反應,PCR) 10 2.2.2 DNA的純化(DNA Purfication) 10 2.2.3 DNA的轉殖(T-A cloning) 10 2.2.4 質體DNA的萃取 11 2.2.5 核苷酸定序(sequencing) 11 2.2.6序列整理與校對 11 2.3 微衛星體引子設計與PCR擴增 12 2.4 變性聚丙烯醯胺膠體電泳分析 12 2.5 資料分析 12 2.5.1遺傳多樣性分析 12 2.5.2 族群間的遺傳歧異度與關係樹的建立 14 2.5.3遺傳單位與空間的關係 14 參、結果 16 一、微衛星基因座在三倍體傅氏鳳尾蕨的廣用性 16 二、微衛星基因座在鳳尾蕨屬物種的廣用性 16 三、二倍體傅氏鳳尾蕨族群遺傳多樣性 16 四、二倍體傅氏鳳尾蕨族群間的遺傳歧異度與族群關係樹 18 五、二倍體傅氏鳳尾蕨族群遺傳單位與空間關係 18 肆、討論 20 一、微衛星基因座的廣用性 20 二、二倍體傅氏鳳尾蕨族群遺傳多樣性與族群結構 20 伍、結論 22 參考文獻 23

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