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研究生: 吳家進
Chia-Chin Wu
論文名稱: 玉山水苦賈及其相關類群之系統分類-特別論及玉山水苦賈的族群遺傳
The systematic study of Veronica morrisonicola and allied species with special reference to the population genetics of Veronica morrisonicola
指導教授: 王震哲
Wang, Jenn-Che
學位類別: 碩士
Master
系所名稱: 生命科學系
Department of Life Science
論文出版年: 2009
畢業學年度: 97
語文別: 中文
論文頁數: 82
中文關鍵詞: 玉山水苦賈系統分類族群遺傳
英文關鍵詞: Veronica morrisonicola, systematic study, population genetics
論文種類: 學術論文
相關次數: 點閱:99下載:3
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  • 玉山水苦賈及其相關類群包含三個在形態上相似的類群,分別為玉山水苦賈(Veronica morrisonicola)、雪山水苦賈(V. tsugitakensis)和玉山前山水苦賈(V. morrisonicola var. yushanchienshanica)。三者皆分佈於高海拔,其中玉山水苦賈分佈最廣,雪山水苦賈的分佈地點侷限在雪山圈谷,而玉山前山水苦賈的分佈地點侷限在玉山前山。本研究利用形質分析和分子資料探討此類群的系統分類問題,特別論及玉山水苦賈的族群遺傳研究。
    本研究利用167個樣本進行形質分析,共測量12個形態特徵。根據ANOVA結果顯示,此三類群在7個外部形態特徵具有顯著差異(葉片長度、葉片寬度、節間長度、總花梗長度、花序長度、花梗長度和果實數量),在歸群分析及PCA分析中可將此三群區分,但仍具有一些中間型的個體,另外玉山水苦賈分佈於合歡山的樣本形態差異較大。由葉綠體DNA所建構的親緣關係顯示,此三類群並非最近緣物種,雪山水苦賈的親緣關係較為疏遠。推估其分歧時間此三類群約為1.36百萬年,玉山水苦賈和玉山前山水苦賈的分歧時間約為0.25百萬年,另外分佈於合歡山的玉山水苦賈雖在形態上差異較大,但分子資料並無差異。
    玉山水苦賈的族群遺傳研究,共使用了142~147個樣本數,合併葉綠體trnL intron和trnL-trnF spacer、核DNA的ITS區域與葉綠體trnD-trnT中的重複片段進行分析。由核DNA結果顯示,基因型歧異度以桃山族群和合歡山族群為最高。檢視基因型關連圖,玉山水苦賈的基因型可分成A、B兩群,其中A群占68%,B群占18.5%。檢視其族群變動歷史顯示,玉山水苦賈的族群數量在A群具有成長趨勢,B群則呈現一個穩定不變的狀態。在族群分化指數中,玉山水苦賈各族群並無顯著分化,由AMOVA檢測呈現族群的變異均在族群內。
    由形質分析和分子資料結果,本研究將Huang and Huang (1998)認定的玉山水苦賈處理為玉山水苦賈(V. morrisonicola)、玉山前山水苦賈(V. morrisonicola var. yushanchienshanica) 和雪山水苦賈(V. tsugitakensis Masamune)。

    Veronica morrisonicola and allied species contains three morphologically similar taxa, namely V. morrisonicola, V. morrisonicola var. yushanchienshanica, V. tsugitakensis. They are distributed in high elevation (2500-3900 m). V. morrisonicola is the widest distribution species. V. tsugitakensis is limited in Hseuhshan. V. morrisonicola var. yushanchienshanica is limited in Yushanchienshan. The present study investigates the systematic of them by using morphological and molecular data special reference to the population genetics of V. morrisonicola.
    In morphometric analysis, total of 167 samples and 12 characters were measured. The results of ANOVA show the seven characters are significantly different between the three taxa. PCA analysis and cluster analysis show that trend to separate into three groups. But in V. morrisonicola there is more variety in population of Hohuanshan. Based on the chloroplast DNA, V. tsugitakensis is most distant from other groups. The study estimate divergent time of the three groups in 1.36 Mya and V. morrisonicola and V. morrisonicola var. yushanchienshanica in 0.25 Mya.
    Population structure analysis of V. morrisonicola were used 142-147 samples. Two chloroplast DNA region (trnL intron and trnL-trnF spacer), nuclear DNA region (ITS) and chloroplast tandem repeat region (trnD-trnT) were used in molecular anlysis. Based on nuclear DNA results, haplotype diversity in population of Taoshan and Hohuanshan are more than others. Based on minimum spaning network, there are two major groups of haplotype in V. morrisonicola. There are 68% in group A and 18.5% im group B. Based on bayesian skyline results indicated that group A was expansion but in group B.
    According morphometric and molecular data, the present study treat V. morrisonicola, V. morrisonicola var. yushanchienshanica, V. tsugitakensis.

    壹、前言………………………………………………………………………………1 一、玉山水苦賈及其相關類群的基本資料………………………………………1 二、研究背景……………………………………………………………………3 三、研究問題…………………………………………………………...……….6 貳、研究材料與方法 (一) 研究材料………………………………………………………………….7 (二) 研究方法……………………………………………………………………8 參、結果……………………………………………………………………………. 19 (一) 玉山水苦賈及其相關類群形質分析及親緣關係……………………13 (二) 分類處理…………………………………………………………………17 (三) 玉山水苦賈之族群遺傳探討…………………………………………17 肆、討論………………………………………………………………………………23 (一) 玉山水苦賈及其相關類群形態比較及親緣關係探討…………………23 (二) 玉山水苦賈形態變異探討………………………………………………25 (三) 玉山水苦賈族群遺傳探討…………………………………………26 伍、結論………………………………………………………………………………29 陸、參考文獻………………………………………………………………………..30 附表目次 表一、玉山水苦賈及其相關類群學名沿革表………………………………………36 表二、玉山水苦賈及其相關類群形態差異比較表…………………………………37 表三、玉山水苦賈形質測量的樣本數量及其採集地點……………………………38 表四、玉山水苦賈分子資料的族群代號及數量……………………………………39 表五、NCBI資料庫中所選用DNA序列的物種編號和名稱………………………40 表六、各序列片段反應條件…………………………………………………………41 表七、PCR反應試藥成分表………………………………………………………42 表八、玉山水苦賈及其相關類群之ANOVA檢測和HSD test…………………….43 表九、玉山水苦賈及其相關類群形態特徵測量平均值比較表……………………45 表十、主成分分析各變數的特徵向量及各軸累積可解釋的變異量………………46 表十一、玉山水苦賈及其相關類群利用核DNA的基因多樣性檢測……………47 表十二A、玉山水苦賈及其相關類群trnL intron片段基因型變異位點組成……48 表十二B、玉山水苦賈及其相關類群trnL-trnF片段基因型變異位點組成………48 表十三、玉山水苦賈ITS片段基因型變異位點組成………………………………49 表十四、玉山水苦賈各族群ITS片段的基因型分佈情況…………………………51 表十五、玉山水苦賈及其相關類群利用葉綠體DNA的基因多樣性檢測………53 表十六、玉山水苦賈各族核DNA的遺傳檢測………………………………54 表十七、葉綠體DNA估算玉山水苦賈各族群之間的族群分化指數(FST)和基因交 流值(Nm)………………………………………………………………..55 表十八、以葉綠體DNA估算玉山水苦賈三大地理區的分化指數(FST)和基因交流 值(Nm)………………………………………………………………55 表十九、核DNA估算玉山水苦賈各族群之間的族群分化指數(FST)和基因交 流值(Nm)………………………………………………………………..56 表二十、以核DNA估算玉山水苦賈三大地理區的分化指數(FST)和基因交流 值(Nm)………………………………………………………………56 表二十一、玉山水苦賈葉綠體DNA的AMOVA檢測表…………………………57 表二十二、玉山水苦賈核DNA的AMOVA檢測表………………………………57 表二十三、玉山水苦賈各族群的葉綠體trnD-trnT片段中重複序列片段的基因型 分佈情況……………………………………………………………………58 表二十四、以葉綠體trnD-trnT片段中重複序列片段估算各族群之間的族群分化指數(FST) ……………………………………………………………………60 表二十五、以葉綠體trnD-trnT片段中重複序列片段估算三大地理區的分化指數(FST)…………………………………………………………………………..60 表二十六、前人研究中所使用的葉綠體DNA核苷酸歧異度比較………………60 附圖目次 圖一、A~L 玉山水苦賈及其相關類群形態特徵測量圖…………………………61 圖二、玉山水苦賈及其相關類群形質分析之主成分分析座標圖…………………65 圖三、玉山水苦賈及其相關類群形質分析之歸群樹分析圖………………………66 圖四、利用葉綠體片段所建構的玉山水苦賈及其相關類群的NJ tree……………67 圖五、利用葉綠體片段所建構的玉山水苦賈及其相關類群的MP tree…………69 圖六、利用葉綠體片段所建構的玉山水苦賈及其相關類群在不同族群中的基因型NJ tree………………………………………………………………………71 圖七、利用葉綠體片段所建構的玉山水苦賈及其相關類群在不同族群中的基因型MP tree………………………………………………………………………72 圖八、玉山水苦賈ITS片段在各族群的基因型分佈圖……………………………73 圖九、玉山水苦賈各族群的葉綠體DNA片段之族群遺傳檢測圖………………74 圖十、玉山水苦賈三大地理區及總樣本數的葉綠體DNA片段之族群遺傳檢測圖……………………………………………………………………………75 圖十一、玉山水苦賈各族群的核DNA片段之族群遺傳檢測圖…………………76 圖十二、玉山水苦賈三大地理區及總樣本數的核DNA片段之族群遺傳檢測圖..77 圖十三、玉山水苦賈的基因型網狀親緣關連圖……………………………………78 圖十四 A ~ C、玉山水苦賈核DNA所有樣本所建構的Bayesian Skyline圖……79 圖十五 A ~ C、玉山水苦賈核DNA,A群所建構的Bayesian Skyline圖…..80 圖十六 A ~ C、玉山水苦賈核DNA,B群所建構的Bayesian Skyline圖…………81 圖十七、玉山水苦賈葉綠體trnD-trnT片段中重複序列片段在各族群的分佈圖..82

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