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研究生:黃偉翔
研究生(外文):Wei-Hsiang Huang
論文名稱:利用葉綠體DNA之atpB及rbcL基因間的非轉譯區間序列探討大陸內蒙古特有種植物四合木(Tetraenamongolica)的親緣地理
論文名稱(外文):Phylogeography of the atpB-rbcL noncoding spacer region of the chloroplast DNA in the endemic species Tetraena mongolica (Zygophyllaceae) in Inner Mongolia
指導教授:劉景煌劉景煌引用關係
指導教授(外文):Zin-Huang Liu
學位類別:碩士
校院名稱:國立中山大學
系所名稱:生物科學系研究所
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2003
畢業學年度:91
語文別:中文
論文頁數:60
中文關鍵詞:親緣網狀圖親緣地理四合木過往棲地破碎化遺傳分化值基因交流值
外文關鍵詞:gene flowpopulation differentiationpast fragmentaionnested clade analysisphylogeographyTetraena mongolica
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中文摘要
本研究藉由葉綠體DNA之atpB及rbcL基因間的非轉譯區間序列來探討四合木 (Tetraena mongolica Maxim.) 的親緣地理型式。四合木是蒺藜科中古老的孓遺單屬種植物,生長在中國內蒙古屬於瀕臨滅絕之特有物種。已列為內蒙古一級保育稀有植物,世界上僅存的四合木分布在內蒙古鄂爾多斯高原的荒漠區,此分布區是亞洲一批古老殘遺植物的避難所。四合木的生殖是以種子繁殖為主要方式,但在惡劣的環境下,則少數併行根部營養繁殖。本實驗共定序出8 個族群77個個體葉綠體DNA之atpB及rbcL基因間的非轉譯區間序列共計880鹼基對。利用葉綠體DNA資料估算族群間的基因交流值 (Nm) (Nm:0.04-2.01) 和族群間分化指數 (Fst) (Fst:0.199-0.895),並將族群間地理距離對數值與分化指數做迴歸分析,結果呈現無顯著相關,顯示四合木其遺傳分化程度不符合「isolation by distance」 模式 (R2=0.0024);根據由葉綠體DNA所構築的親緣網狀圖(minimum spanning network),檢測地理形式與clade間的關係,顯示四合木族群地理區所呈現的地理分布模式,主要是受到過去棲地破碎化 (past fragmentation) 的影響,而黃河氾濫及千里山山脈的阻隔可能是造成四合木地理隔離的主因。
Abstract

Sequence variation of the atpB-rbcL noncoding spacer region of the chloroplast DNA was used to reconstruct the phylogeography of Tetraena mongolica.T. mongolica (Zogophylaceae) is a relict species of a monotypic genus of in Inner Mongolia of China. It has been listed as seriously endangered plants that only distributed on the Ordos plateau of Inner Mongolic. The area is a refuge of many relict Asian Plants. Reproduction of T. mongolica is mainly by seed but a reproduction is anther way under the stress. A total of 880 bp of the atpB-rbcL spacer of cpDNA was sequenced from 8 populations of T. Mongolic. According to the cpDNA data, the level of gene flow (Nm:0.04-2.01), and population differentiation (Fst:0.199-0.895) were indicated, and the “isolation by distance” model was not supported (R2 =0.0024). Based on minimum spanning network and nested clade analysis, the past fragmentation is the main phylogeographic factor affecting the population structure of T. mongolica .We suggested the deluges of Yellow River and isolation of Gianlishan Mountain cased the population fragmentation of T. mongolica.

Keywords: phylogeography, Tetraena mongolica, gene flow, population
differentiation, nested clade analysis, past fragmentation
目錄
誌謝................................................................................................. Ⅰ
中文摘要…………………………………………………………..Ⅱ
英文摘要…………………………………………………………..Ⅲ
目錄………………………………………………………………..Ⅳ
表目錄……………………………………………………………..Ⅵ
圖目錄……………………………………………………………..Ⅶ

第壹章 前言……………………………………………………….. 1
一、親緣地理學的研究………………………………………… 1
二、四合木的研究歷史………………………………………… 5
三、瀕危物種的保育與其遺傳變異…………………………...11
四、分子技術進步對於族群結構的應用……………………...15
五、研究目的 ………………………………………………….16
第貳材料與方法…………………………………………………...17
一、研究材料...............................................................................17
二、實驗方法 ........................................……………………….18
第參章 結果....…………………………………………………….24
一、葉綠體DNA atpB-rbcL noncoding spacer片段……….24
二、四合木的親緣地理………………………………………...26
第肆章 討論……………………………………………………….27
一、四合木的遺傳變異………………………………………...27
二、四合木的族群分化………………………………………...28
三、四合木的親緣地理模式…………………………………...29
四、四合木的保育……………………………………………...31
第伍章 結論……………………………………………………….32
第陸章 參考文獻…………………………………………………..33
表……………………………………………………………………40
圖……………………………………………………………………47
附錄…………………………………………………………………57

表目錄
表一、四合木之採樣地點、採集個體數目及族群代號.................40
表二、進行四合木DNA介於atpB及rbcL基因間的noncoding
spacer聚合酵素連鎖反應所使用之引子序列……………41
表三、根據葉綠體DNA資料估算四合木族群haplotype歧異
度、核柑酸歧異度………………………………………… 42
表四、根據葉綠體DNA資料所建構樹狀圖中各clade的地理
族群組成…………………………………………………….43
表五、根據葉綠體DNA資料估算四合木族群間基因交流值和
族群間分化指數…………………………………………….44
表六、估算葉綠體DNA親緣網狀圖各clade的Dc、Dn值……..45
表七、根據所估算之葉綠體DNA親緣網狀圖各clade的Dc、
Dn值,並配合檢索表推測四合木不同的clade所曾遭
遇的歷史事件……………………………………………….46

圖目錄
圖一、四合木植株照片....................................................................47
圖二、四合木之形態特徵…………………………………………48
圖三、四合木之取樣分布圖………………………………………49
圖四、四合木葉綠體DNA之atpB及rbcL 基因間的非轉譯區
間區列在八個族群的代表序列……………………………50
圖五、根據葉綠體DNA資料以聚分類分析法構築四合木的親
緣樹狀圖……………………………………………………54
圖六、根據葉綠體DNA資料構築的親緣網狀圖…………………55
圖七、四合木族群地理距離對數值與利用葉綠體DNA資料估
算的遺傳分化值之迴歸分析圖……………………………56
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