臺灣博碩士論文加值系統

石蓴科為廣泛分佈於世界各地沿岸的大型綠藻，台灣沿岸過往的石蓴科藻種紀錄有二屬十三種，依藻體外觀形態可分為兩型：1）葉片型－藻體由雙層細胞組成，同一藻種外觀多變，呈長條帶狀、小株牡丹狀與膜狀等；2）管狀型－藻體由單層細胞組成中空管狀分支。在過去，台灣產石蓴科的分類研究，大多以外觀形態進行分類鑑定，易造成分類混淆或同種異名。因此，本研究除觀察藻體外觀與營養結構外，並利用核酮糖二磷酸羧化酶-加氧酶大次單元序列進行台灣產石蓴科的系統分類及親源關係探討。分子序列分析結果顯示台灣產石蓴科共有二屬十三個藻種，其中，“緣管滸苔”、“滸苔” 及 “裂片石蓴” 為台灣已記錄藻種，奄美墨石蓴、台灣墨石蓴、“大野石蓴” 及 “刺狀石蓴” 為新紀錄種。依分子序列結果可將台灣產石蓴科二屬十三種歸為三大演化群：1）“石蓴屬管狀型” 藻種演化群，此大演化分群中來自台灣有七種，形態外觀由單層細胞所組成中空管狀結構，其中包含二種台灣已紀錄種（“緣管滸苔” 和 “滸苔”），其它五種為未確定種（“石蓴屬” sp. 1、“石蓴屬” sp. 2、“石蓴屬” sp. 3、“石蓴屬” sp. 4 和 “石蓴屬” sp. 5）；2）石蓴屬/墨石蓴屬/雙葉藻屬/狹葉藻屬/苔藻屬複合演化群，此大演化分群中，台灣具有三個藻種，其中包含一種台灣新紀錄種（奄美墨石蓴）、一種世界新種（台灣墨石蓴）及一種為未確定種（石蓴屬 sp. 6），石蓴屬 sp. 6 為單層細胞組成管狀結構，而墨石蓴屬外觀為雙層細胞葉片膜狀結構，顏色為墨綠色或橄欖色，觸感粗厚；3）“石蓴屬葉片型” 藻種演化群，此大演化分群中，台灣具有三個藻種，形態外觀為雙層細胞葉片型結構（“裂片石蓴”、“大野石蓴” 及 “刺狀石蓴”），顏色為草綠色或淺綠色。雖然分生結果不支持石蓴屬為單一系源體系，但因目前只用了單一基因序列分析，故本研究暫以形態差異來定義石蓴屬，以引號來標記沒有與石蓴屬模式種位於同一演化分支的藻種。另一方面，由研究分子序列結果及形態觀察推論過去文獻紀錄的 “牡丹石蓴” 可能為 “刺狀石蓴” 的誤判，而台灣產的 “石蓴” 應為 “大野石蓴” 或 “刺狀石蓴” 二藻種的誤判。分生結果亦顯示台灣之前紀錄為 “孔石蓴”、“硬石蓴” 和 “日本墨石蓴” 的核酮糖二磷酸羧化酶-加氧酶大次單元序列與來自日本的奄美墨石蓴相同。另外，本研究發現的六個石蓴屬未確定藻種可能為世界新種，但需與其它海域相似藻種進行詳細比較之後，才能確認。來自南台灣香蕉灣的台灣墨石蓴，在親源關係樹狀圖中並無相對應序列，因此將此藻種判定為世界新種。然而，本研究結果中並無顯示過去所記錄藻種共六種（條滸苔、扁石髮、腸石髮、狹葉石蓴及網石蓴），其中因本實驗室沒有能進行去氧核醣核酸定序之網石蓴標本，導致結果並未顯示此藻種，而其他這些藻種過去可能因為形態外觀上的混淆，而導致同種異名的情況發生。

Two genera and thirteen species of the Ulvaceae have been recorded from Taiwan. Based on thallus morphology, the species can be grouped into two types: 1) Blade-type: thalli composed of two-celled layers; 2) Tubular-type: thalli composed of a single celled layer. In the past, the taxonomic studies on the Ulvaceae from Taiwan were largely based on morphological observations resulting some taxonomic confusion and synonymy. In this study, the taxonomy of the Ulvaceae is revised based on comparative anatomy and the inter-specific relationships are inferred using rbcL sequence analyses. The molecular analyses showed that there are thirteen species identified, which are grouped into three, evolutionary distinct clades: 1) tubular-type “Ulva” clade: containing seven species, “U.” prolifera and “U.” linza and five undetermined species (“Ulva” sp. 1, “Ulva” sp. 2, “Ulva” sp. 3, “Ulva” sp. 4 and “Ulva” sp. 5); 2) Ulva / Umbraulva/Gemina/Percursaria/Ulvaria complex: composed of three species, Um. amamiensis, Um. taiwanensis sp. nov. and one undetermined species with tubular form (Ulva sp. 6); 3) blade-type “Ulva” clade: including three species, “U.” fasciata, “U.” ohnoi and “U.” spinulosa, which thalli are in either grass-green or in light-green.
The previous records “Ulva” prolifera, “U.” linza and “U.” fasciata from Taiwan are confirmed, whereas one new species of Umbraulva, Um. taiwanensis is discovered and Umbraulva amamiensis, “U.” ohnoi and “U.” spinulosa are newly recorded from Taiwan. On the other hand, the previous record of “U. conglobata” could be “U.” spinulosa, whereas “U. lactuca” is not present in Taiwan and is a mixture of “U.” ohnoi and “U.” spinulosa. The molecular analyses also indicated that “U. pertusa”, “U. rigida” and “Um. japonica” are not occurring in Taiwan, but are molecularly identical to Um. amamiensis from Japan. In addition, the study found that the six undetermined species may be new species of the world, but need other species of seaweeds detailed comparisons. The five species, U. clathrata, U. compressa, U. intestinalis and U. angusta, are not detected in this study and can be misidentification of other species. The distribution of the species of the Ulvaceae found in Taiwan and their phylogenetic relationships with related species worldwide are also discussed in this study.

目錄

謝辭………………………………………………………………………………….....I
摘要…………………………………………………………....…………..………….II
Abstract…………………………………………………………….……...…………III
目錄………………………………………………..………….…………………...…IV
圖目錄………………………………………………..………….………………...…VI
表目錄…………………………………………..……………….……………....…VIII
綠藻相關專有名詞解釋……………………………………..………………………IX
第一章、前言………………………………………………….……….……………..1
1.1、石蓴科藻種簡介………………………………………………..….………1
1.2、石蓴科藻種分類研究歷史…………………………………………………1
1.3、台灣產石蓴科藻種研究歷史與分類情況……………………………...…3
1.4、石蓴科藻種生活史………….………....……………………..……………5
1.5、研究動機與目的………………………….…………..……………………6
第二章、材料與方法…………………………………………………………………7
2.1、藻種採集與保存……………………………………...……………………7
2.1.1、蠟葉標本製作過程…………………………………………..……….9
2.2、分子序列分析方法……………………………………………………….…9
2.2.1、去氧核醣核酸萃取（DNA Extraction）………………………………..9
2.2.2、基因擴增（Gene Amplification）：聚合酶連鎖反應（Polymerase Chain Reaction；PCR）……………………………………………………........…10
2.2.3、定序反應及基因定序（Sequencing Reaction and Gene Sequencing）
………………………………………………………………………………11
2.2.4、親源關係分析（Phylogenetic Analysis）………………....……….11
2.3、形態觀察…………………………………….……………………………12
2.3.1、染劑配製………………………………….……………..………….12
2.3.2、切片染色方法…………………………………….…..…………….13
第三章、結果……………………………………………………….……………….15
3.1、分子親源關係樹分析………………………………………………….…15
3.2、形態觀察結果…………………..………………………………………….23
3.2.1、台灣產石蓴科（Ulvaceae Lamouroux ex Dumortier 1822 : 72, 102）
……………………………………………………………………….......….24
3.2.2、台灣產墨石蓴屬（Umbraulva Bae &; Lee 2001 : 230）…….……24
3.2.2.1、Umbraulva amamiensis (Tanaka) Bae &; Lee………………25
3.2.2.2、Umbraulva taiwanensis T.-Y. Huang &; S.-M. Lin sp. nov.…30
3.2.3、台灣產石蓴屬（Ulva Linnaeus 1753 : 1163）…………….....……35
3.2.3.1、“Ulva” linza Linnaeus…….…………………………………37
3.2.3.2、“Ulva” sp. 1…….………………..………………….………43
3.2.3.3、“Ulva” prolifera O. F. Müller…….…………………………48
3.2.3.4、“Ulva” sp. 2…….……………………………….……………53
3.2.3.5、“Ulva” sp. 3………………………………………………....58
3.2.3.6、“Ulva” sp. 4…………….………………..……………….…63
3.2.3.7、“Ulva” sp. 5…….……………………………………….……68
3.2.3.8、Ulva sp. 6……………………………………………………73
3.2.3.9、“Ulva” fasciata Delile……………….……..…………….…77
3.2.3.10、“Ulva” ohnoi Hiraoka &; Shimada………………………...84
3.2.3.11、“Ulva” spinulosa Okamura &; Segawa…….……...……….89
第四章、討論與結論………………………………………………………………..97
第五章、參考文獻……………………………………………………………...….105

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