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研究生:謝鎮宇
研究生(外文):Chen-Yu Hsieh
論文名稱:利用LEAFY基因序列探討台灣玉山杜鵑複合群的起源及演化
論文名稱(外文):The origin and evolution history of Rhododendron species complex in Taiwan based on the intron sequences of LEAFY gene
指導教授:黃士穎
指導教授(外文):Shih-Ying Hwang
學位類別:碩士
校院名稱:中國文化大學
系所名稱:生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:72
中文關鍵詞:玉山杜鵑複合群演化起源親緣地理
外文關鍵詞:Rhododendron pseudochrysanthum species complexLEAFY genephylogeographyorigin
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杜鵑花,屬於杜鵑花科 ( Ericaceae ) 杜鵑花屬 ( Rhododendron ) 植物,廣泛分佈於全球除了非洲及南美之外的地方,在台灣則有16種原生杜鵑生長在全台各地,其中紅星、玉山、森氏及南湖杜鵑則以中高海拔山區為其主要分佈範圍。台灣高山原生杜鵑中,紅星、玉山、森氏杜鵑由於在花期、型態、分佈均有不同,因此在分類上有很大的爭議,在台灣植物誌第四版中以傳統分類的觀點將紅星、玉山、森氏及南湖杜鵑歸為玉山杜鵑一個種,但是在分子證據上則有不同的看法,因此在研究中將紅星、玉山、森氏及南湖杜鵑統稱為玉山杜鵑複合群。
在傳統分類上,學者認為紅星與森氏是由玉山杜鵑演化而來,但在cpDNA的trn L-trn F、atp B-rbc L基因片段分析中並沒有觀察到這樣的演化結果,且cpDNA的單套型分析結果僅觀察到紅星杜鵑演化成玉山與森氏杜鵑,與傳統分類學者所提出的說法相異。此外四種原生杜鵑彼此間的基因歧異度低,推測可能是由於過去冰河時期廣泛分佈於中低海拔,基因交流頻繁的關係,而在冰河期結束後氣溫漸漸回升,使杜鵑受到population fragmentation,殘存的族群由中低海拔往中高海拔山區移動,才慢慢演化為今日所見之結果。
過去學者提出假設,認為玉山杜鵑複合群為單一起源,且在冰河時期是廣泛分佈於中低海拔,因此本研究以杜鵑細胞核之LEAFY基因第二段intron序列做為分析材料,希望能了解玉山杜鵑複合群的演化關係,藉此了解其演化支系以供保育及復育之用,並找出證明玉山杜鵑複合群曾經廣泛分佈於中低海拔的分子證據,而本研究結果亦符合此假設的說法,在演化關係上發現玉山杜鵑複合群的起源應為玉山杜鵑,且森氏、南湖杜鵑亦應併入玉山杜鵑之中,而紅星杜鵑則有逐漸分化為其他物種的趨勢。
The genus Rhododendron is distributed throughout the world except Africa and South America. In Taiwan, Rhododendron species are found from the lowlands to the highest mountain. But Taiwanese Rhododendron species, which are only finding in middle to high elevation, contain four species. These four Taiwanese Rhododendron species include R.hyperythrum、R.rubropanctatum、R.morii and R.pseudochrysanthum. In these species, taxonomists classified them as a single species (R.pseudochrysanthum) by morphological way (Flora of Taiwan ver.4).
In fact, there is only a 0.4% genetic distance (Kimura’s two-parameter distance) found between R.rubropanctatum and R.hyperythrum using nuclear ITS (internal transcribed sequence) sequences. This evidence supports the morphological observations that R.hyperythrum should be combined with the R.rubropanctatum species called R.rubropanctatum complex.
Taxonomists in Taiwan regard that both R.rubropanctatum and R.morii as being derived from R.pseudochrysanthum. But there is no evidence support the opinion in the research of cpDNA trnL-trnF and atpB-rbcL sequence. By the way, the research supports the once-widespread distribution of the R.rubropanctatum complex at middle elevations. And the low genetic diversity of Taiwanese Rhododendron cpDNA trnL-trnF and atpB-rbcL sequences may due to population fragmentation caused by rising temperature in the post-Pleistocene.
In this study, we use rapidly evolving nuclear gene, LEAFY gene, in order to know the evolution history and the range expansion of R.pseudochrysanthum complex in the post-Pleistocene. Our reasult not only find the evidence, which can support the hypotheses, but also find that, R.pseudochrysanthum, is the origin of the R.pseudochrysanthum complex.
壹、前言 1
貳、前人研究 3
參、材料與方法 14
一、材料收集與樣本處理 14
二、DNA萃取、定量與稀釋 14
三、聚合酶連鎖反應 17
四、Subcloning 19
五、Colony PCR / sequencing 20
六、資料分析 21
1.序列分析及基因譜系分析 21
2.遺傳變異分析 22
(1)單套型歧異度( h) 22
(2)核苷酸歧異度(π) 23
(3)核苷酸分歧度(θ) 23
3.中性假說測試 23
4.族群分化指數及基因流傳值分析 24
5.族群間親緣結構分析 25
肆、結果 27
伍、討論 33
陸、結論 40
柒、參考文獻 41
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