透過粒線體基因體探討盤古蟾蜍與中華蟾蜍在蟾蜍科之內的親緣關係

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黃渝雯(Yu-Wen Huang) 查詢紙本館藏

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論文名稱

透過粒線體基因體探討盤古蟾蜍與中華蟾蜍在蟾蜍科之內的親緣關係
(Exploring the genetic relationship between Bufo bankorensis and Bufo gargarizans within the family Bufonidae through the mitochondrial genome)

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至系統瀏覽論文 (2028-7-21以後開放)

摘要(中)

在全世界的蟾蜍科裡共有五十三屬，六百四十二種的蟾蜍。在台灣僅見兩種原生種蟾蜍，分別為盤古蟾蜍（Bufo bankorensis）與黑眶蟾蜍（Duttaphrynus melanostictus）。先前研究透過大量形態特徵的分析、cytb限制酶切割實驗及粒線體基因ND2與D-loop序列親緣關係樹分析，認為盤古蟾蜍為台灣特有種，與中華蟾蜍為不同物種，但有部分學者透過粒線體DNA基因(12S、16S)分析，認為盤古蟾蜍與中國的中華蟾蜍(Bufo gargarizans)為同種異名，因此盤古蟾蜍與中華蟾蜍的分類地位，目前尚有爭議。
本研究使用從台灣陽明山採集的盤古蟾蜍，進行粒線體基因體DNA定序，同時收集了已發表的十六種蟾蜍(十九條)擁有粒線體基因體的序列，以及中華蟾蜍物種複合群與姊妹種的五個基因序列(12S、16S、COI、cytb及ND2)，透過分子系統、演化生物學分析及遺傳距離的比較，認為中華蟾蜍物種複合群內已發表的物種皆為有效物種，是具有分類上的意義；而盤古蟾蜍在台灣分為東、西兩群，西台灣的盤古蟾蜍與安徽的中華蟾蜍為同一演化支，關於盤古蟾蜍是否為台灣特有種，仍需要增加東台灣的資料進被夜市討。

摘要(英)

There are 642 species of toads in 53 genera around the whole world. 16 species of toads have been published their complete mitochondrial genomes. The only two native species of toads found in Taiwan were the Bufo bankorensis and Duttaphrynus melanostictus. Some previous studies believed that the Bufo bankorensis was a distinct endemic species in Taiwan by analyses of numerous morphometric characters, cytb restriction enzyme cutting experiments and phylogeny tree analysis of mitochondrial genes ND2 and D-loop sequences.. However, some scholars have suggested that the Bufo gargarizans and the Bufo bankorensis were the same species through several mitochondrial DNA gene (12S, 16S) analyses.
This study used Bufo bankorensis collected from Yangmingshan, Taiwan, to sequence the complete mitochondrial genes. At the same time, the published sequences of sixteen toad species (nineteen sequences) possessing complete mitochondrial genes were collected, as well as the five gene sequences (12S, 16S, COI, cytb and ND2) of the Bufo gargarizans species complex and other toad sister species, through molecular systems, evolutionary biology analyses and p-distances, genetic distances. Our data showed that all published species in the Bufo gargarizans species complex were effective species with classification significance. Bufo bankorensis were divided into eastern and western groups in Taiwan. Bufo bankorensis of western Taiwan and Bufo gargarizans from Anhui belong to the same clade. As for whether Bufo bankorensis endemic to Taiwan, it was still required more data from the eastern Taiwan group.

關鍵字(中)

★ 盤古蟾蜍
★ 粒線體基因體
★ 中華蟾蜍
★ 中華蟾蜍物種複合群
★ 貝葉氏推斷法
★ 最大概率演算法

關鍵字(英)

論文目次

目錄
摘要 i
Abstract ii
致謝 iii
目錄 iv
圖目錄 v
表目錄 viii
縮寫檢索表 ix
一、緒論 1
1.1物種的定義…………………………………………………………...1
1.2兩棲類 2
1.3盤古蟾蜍 3
1.4粒線體和粒線體DNA(mitochondrial DNA，mtDNA) 5
1.5中華蟾蜍物種複合群(Bufo gargarizans species complex) 5
1.6粒線體基因體在物種分類上的應用 6
二、材料與方法 8
2.1實驗樣品 8
2.2 DNA粗萃取 8
2.3引子設計 9
2.4聚合酶連鎖反應(PCR，polymerase chain reaction) 10
2.5 PCR 產物電泳 11
2.6 DNA 純化 11
2.7 DNA 定序(sequencing) 12
2.8整理序列 12
2.9 DNA 序列及演化分析 12
2.10中華蟾蜍物種複合群(Bufo gargarizans species complex) 14
2.11姊妹種(sister species) 15
三、結果 16
3.1盤古蟾蜍粒線體基因體 16
3.2親緣關係分子分析結果 20
四、討論與結論 26
4.1 D-loop重複序列 26
4.2中華蟾蜍物種複合群的假說是否成立 26
4.3目前粒線體基因體資料所呈中華蟾蜍物種複合群的親緣關係 27
4.4盤古蟾蜍是否為台灣特有種 28
五、參考文獻 29
附錄一 34
附錄二 40
附錄三 43

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指導教授

劉阜果(Fu-Guo Robert Liu)

審核日期

2024-7-26

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