面天樹蛙全粒線體基因組序列與其親緣關係

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曾明彥(MING-YEN TSENG) 查詢紙本館藏

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論文名稱

面天樹蛙全粒線體基因組序列與其親緣關係
(Mitochondrial genome sequence of Kurixalus idiootocus and phylogenetic analyses)

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至系統瀏覽論文 (2025-12-25以後開放)

摘要(中)

全台灣的樹蛙有4屬14種，面天樹蛙(Kurixalus idiootocus)屬於其中的一種，分佈於台灣中低海拔氣候潮濕的灌木叢或草原。目前親緣關係學的相關研究只有利用部分粒線體基因片段來探討親緣關係的遠近，並無全粒線體基因組完整序列的研究。因此，本研究使用從宜蘭採集的樣本，利用聚合酶連鎖反應技術(PCR)擴增全粒線體基因組完整序列加以定序，序列總長度為20,944 bp，包含37個基因。此資料存放於NCBI GenBank中，登錄號為MZ636491，粒線體全基因組核苷酸組成比例為: 腺嘌呤(Adenine, A) 30.7%、胸腺嘧啶(Thymine, T) 32.0%、胞嘧啶(Cytosine, C) 23.5%、鳥嘌呤(Guanine, G) 13.8%。本研究發現面天樹蛙與其他樹蛙科青蛙皆有發生ND5 基因重新排列的情況，此外，最大簡約法(Maximum parsimony, MP)、貝葉氏推斷(Bayesian Inference, BI）、鄰近連接法(Neighbor-joining, NJ)和最大概率估計(Maximum Likelihood, ML)的分析結果皆顯示，相較於溪樹蛙屬(Buergeria)，面天樹蛙跟樹蛙屬(Rhacophorus)和泛樹蛙屬(Polypedates)關係較近。目前而言，本研究的面天樹蛙全粒線體基因組完整序列是原指樹蛙屬(Kurixalus)中第一筆全粒線體序列資料，此研究結果可進一步提供於親緣關係學與基因體學的研究。

摘要(英)

There are 4 genera 14 species of Family Rhacophoridae in Taiwan, including Kurixalus idiootocus. This species distributes in the middle and low altitude humid shrublands or grasslands of Taiwan. At present, phylogenetic studies related to Kurixalus idiootocus are based on partial mitochondrial gene sequences and no its whole mitochondrial genome information. Therefore, this research subject used a sample collected from Yilan and conducted the polymerase chain reaction (PCR) technique to amplify the sequence of its whole mitochondrial genome for sequencing. The total length of it was 20,944 bp, which contained 37 genes. This data was deposited in NCBI GenBank with the accession number MZ636491. The base composition of the complete mitochondrial DNA was 30.7% for A, 32.0% for T, 23.5% for C, and 13.8% for G. This study found Kurixalus idiootocus and other Rhacophoridae frogs all had ND5 gene rearrangement. Phylogenetic relationships with Bayesian Inference, Maximum Likelihood, Maximum parsimony, and Neighbor-joining all suggested the phylogenetic relationship of Kurixalus idiootocus close to genera Rhacophorus and Polypedates rather than Buergeria. Currently, the whole mitochondrial genome sequence of Kurixalus idiootocus was the first data about the whole mitochondrial genome sequence in genus Kurixalus. This research results could be used for further studies in the field of phylogenetics and genomics.

關鍵字(中)

★ 面天樹蛙
★ 親緣關係學
★ 粒線體DNA

關鍵字(英)

★ Kurixalus idiootocus
★ phylogenetic analyses

論文目次

摘要……………………………………………………………………………..i
Abstract…………………………………………………………………………ii
致謝……………………………………………………………………………..iii
目錄……………………………………………………………………………..iv
圖目錄…………………………………………………………………………..vi
表目錄…………………………………………………………………………viii
縮寫檢索表……………………………………………………………………...x
一. 緒論…………………………………………………………………………1
1.1 面天樹蛙………………………………………………………………...1
1.2 粒線體…………………………………………………………………...3
1.3 分子生物技術…………………………………………………………...6
1.4 面天樹蛙(Kurixalus idiootocus)研究回顧……………………………..8
1.5 研究目的……………………………………………………………….11
二. 材料與方法………………………………………………………………..12
2.1 樣本採集……………………………………………………………….12
2.2 DNA萃取……………………………………………………………....12
2.3引子(Primer)設計……………………………………………………...13
2.4 聚合酵素鏈鎖反應(Polymerase chain reaction, PCR) ……………..16
2.5 DNA純化(DNA purification) ………………………………………...17
2.6 DNA定序(DNA sequencing) ………………………............................17
2.7 整合序列(Sequence alignment) …………….......................................17
2.8 分子系統及演化生物學分析(Phylogenetic Analysis) ……………....17
三. 結果………………………………………………………………………..21
3.1 粒線體全基因組大小與組成………………………………………….21
3.2 蛋白質基因(Protein-coding genes) …………………………………..27
3.3 非編碼基因(non-coding genes) ………………………………………38
3.4 D-loop與The replication origin of the L-strand replication (OL)…46
3.5 親緣關係樹(Phylogenetic Tree)分析……………………………........55
四. 討論………………………………………………………………………..60
五. 結論………………………………………………………………………..63
六. 參考文獻…………………………………………………………………..65
附錄一………………………………………………………………………….73

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楊懿如 (2005)“臺灣兩棲動物野外調查手册”，行政院農業委員會林務局出版.

指導教授

劉阜果(Fu-Guo Robert Liu)

審核日期

2021-12-28

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