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研究生:吳念真
研究生(外文):Nien-Chen Wu
論文名稱:台灣地區蜉蝣目昆蟲的COI基因組成及其地理區系變異之探討
論文名稱(外文):The COI sequence composition and geographic variation of Ephemeroptera in Taiwan
指導教授:葉文斌葉文斌引用關係
指導教授(外文):Wen-Bin Yeh
學位類別:碩士
校院名稱:國立中興大學
系所名稱:昆蟲學系所
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:2007
畢業學年度:96
語文別:中文
論文頁數:72
中文關鍵詞:粒線體細胞色素氧化酶第一次單元台灣蜉蝣目地理區系變異
外文關鍵詞:cytochrome oxidase I (COI)TaiwanEphemeropterageographic variation
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本研究利用粒線體細胞色素氧化酶第一次單元(cytochrome oxidase I, COI) 核苷酸序列作為分子標誌,透過DNA層面的資訊來釐清台灣地區受中央山脈分隔的東部及西部各個流域間,蜉蝣目的分化情形及遷徙路徑,也同時建立台灣地區部份蜉蝣種類(7科36種)之DNA序列資料庫,加速隱匿種的分類鑑定。結果顯示,COI序列在種內的變異多數小於0.025,種間及屬間的變異的變異大多介於0.18及0.21之間,而科間的變異則多大於0.18,最高達0.31。親緣分析顯示,此基因並無法界定科間的親緣關係。但在Heptageniidae科的Afronurus floreus及A. chihpenensis兩個形態種的種內分化程度相當大,明顯分成三個支系;Rhithrogena屬大略可劃分為七個小支系,但分析的兩個種類並未明確的區分開來。地理親緣關係的結果顯示台灣的蜉蝣物種,有藉由南部地區遷移至東部地區的現象:像A. floreus在台灣可能最先分佈於台灣西半部,後經由南部再向上遷移至東部;Baetiella bispinosa很早之前就由南部上遷至東部,形成目前的群間高度分化情形。分析結果也可推測Baetidae 科Group11、Group12的東、西部族群應在南部地區形成雜交區。此外,除A. floreus, A. chihpenensis及幾個高度分化的種群外,Rhithrogena屬的R. ampla與R. parva兩個種無論是在親緣樹或是單倍型網狀圖中,皆無法明確的將兩個種區分開來,使得此這些蜉蝣物種的分類地位均值得更深入的探討。
Sequences of cytochrome oxidase I were used to elucidate the genetic differentiation and the immigrant possibility of mayflies populations distributed either in eastern or western Taiwan; and also to address the species identification. Mostly, sequences variations were less than 0.025 within species, 0.18-0.21 between species and genus in the same family, 0.18-0.31 between families. Phylogenetic analysis revealed that resolution of COI sequences was low in family relationships. Three phylogenetic lineages were found both in true morphological species, i. e. Afronurus floreus and A. chihpenensis, of Heptageniidae. Whereas, true distinct morphological Rhithrogena species were intermingled in phylogenetic tree and grouped into seven separate lineages. Results of phylogeographical analysis show that many mayflies migrated from western Taiwan to eastern Taiwan via area of southern Taiwan. Individuals of A. floreus with high diversity in western populations and migrated to Taitung and Hualien counties via southern Taiwan recently. Individuals of Baetiella bispinosa moved long time ago, and led to many isolated populations in eastern Taiwan. Although high diversity occurred within the species of A. floreus, A. chihpenensis and some Baetidae species group, the species recognition of medley COI lineages of true morphological Rhithrogena species were necessary to be described.
目錄
誌謝 i
中文摘要 ii
英文摘要 iv
目錄 v
一、前言 1
二、前人研究 3
三、材料與方法 8
(一)、標本來源 8
(二)、標本鑑定 8
(三)、DNA萃取 8
(四)、PCR增幅目標DNA 9
(五)、電泳分析 10
(六)、PCR產物純化 10
(七)、定序 11
(八)、DNA序列之整理及排序 11
(九)、核苷酸變異分析 12
(十)、親緣關係樹之建立 12
(十一)、親緣地理學之研究 13
四、結果 14
(一)、分析之物種種類 14
(二)、分類群序列變異分析 14
(三)、各密碼位變異分析 16
(四)、親緣關係樹分析 16
(五)、各物種地理的變異 17
五、討論 22
(一)、界定各形態種或支系的DNA變異物種-分類位階的變動 22
(二)、親緣關係樹 22
(三)、各物種地理親緣關係 24
六、參考文獻 26


表次索引
表一、本試驗中所有蜉蝣明細及採集資訊 34
表二、蜉蝣COI序列核苷酸百分比組成 51
表三、各科科內屬間、屬內種間及種內的變異範圍 57
表四、個體數量大於一的同物種其序列變異範圍 58

圖次索引
圖一、蜉蝣採集地點位置分佈圖 59
圖二、蜉蝣COI序列間變異的百分比分佈圖 60
圖三、蜉蝣各分類階層COI序列間變異範圍的數量及分佈 61
圖四、各科科內的變異範圍分佈比較圖 62
圖五、蜉蝣樣本間的COI DNA序列差異總變異(Tvs)與其相對的顛換取代(Tv)及轉換取代(Ts)之散佈圖 63
圖六、蜉蝣COI區段以p-distance運算模式再經鄰接法所演算出之親緣關係樹 64
圖七、Heptageniidae科Afronurus floreus所有個體之COI區段單倍型之最大簡約網狀圖及親緣關係樹 67
圖八、Heptageniidae科Rhithrogena屬2種R. ampla及R. prava所有個體之COI區段單倍型之最大簡約網狀圖及親緣關係樹 68
圖九、Baetidae科Acentrella lata所有個體之COI區段單倍型之最大簡約網狀圖及親緣關係樹 69
圖十、Baetidae科Baetiella bispinosa所有個體之COI區段單倍型之最大簡約網狀圖及親緣關係樹 70
圖十一、Baetidae科Group 11所有個體之COI區段單倍型之最大簡約網狀圖及親緣關係樹 71
圖十二、Baetidae科Group 12所有個體之COI區段單倍型之最大簡約網狀圖及親緣關係樹 72
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