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研究生:林文謙
研究生(外文):Lin, Wen-Chien
論文名稱:臺灣產海洋紅藻海膜藻屬(海膜藻科,紅藻門)分子親源關係與分類研究
論文名稱(外文):Taxonomic revision and molecular phylogeny of the marine red algal genus Halymenia (Halymeniaceae, Rhodophyta)from Taiwan
指導教授:林綉美林綉美引用關係
指導教授(外文):Lin Showe-Mei
口試委員:彭家禮、劉少倫、林綉美
口試日期:2014-07-01
學位類別:碩士
校院名稱:國立臺灣海洋大學
系所名稱:海洋生物研究所
學門:自然科學學門
學類:海洋科學學類
論文種類:學術論文
論文出版年:2014
畢業學年度:102
語文別:中文
論文頁數:116
中文關鍵詞:分類、分子親源關係、臺灣、海膜藻科、海膜藻屬、杜氏海膜
外文關鍵詞:taxonomy、molecular phylogeny、Taiwan、Halymeniaceae、Halymenia、Halymenia durvillei
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海膜藻屬為廣泛分佈於全球熱帶、亞熱帶、溫帶海域的大型紅藻,通常生長於低潮線至潮下帶之礁石上。臺灣本島與離島沿海全年可見,但西部沿海的紀錄卻相當稀少,其生長最旺盛的季節是春季至初夏。海膜藻屬目前世界上紀錄共69種,藻體依據分子親源關係與外觀形態觀察主要可分為兩種型態:羽狀分枝型以及匍匐葉狀型。海膜藻屬特徵為藻體呈暗紅色、柔軟黏滑、具盤狀固著器。過去的臺灣學者大多會將藻體的外觀形態特徵做為分類鑑定的基礎,但若只憑外觀進行鑑定,將可能導致同種異名或同名異種的混淆。因此以海膜藻屬為研究對象,除進行藻體內外形態特徵比較外,並配合二磷酸羧化酶–加氧酶大次單元序列來探討臺灣產海膜藻屬與各地海膜藻科的分子親源關係。分子序列分析結果顯示臺灣海膜藻屬有四種不同的分化群(=藻種),當中杜氏海膜為臺灣過去已記錄種,湯都海膜為臺灣新紀錄種,“葉狀海膜–1"、“葉狀海膜–2"可能為世界新種。由於與有些已知紀錄於西太平洋或其它海域的海膜藻種外觀十分相像,臺灣海膜藻種易被誤認為是已知的藻種,但若是透過形態特徵配合親源關係分析來進行比較,方可區分出藻種。從臺灣產的四種海膜藻種依據分子親源關係與外觀形態觀察發現:1)分子親源關係的分析結果顯示,臺灣過去所紀錄的“小果海膜"、“海膜"、“錫蘭海膜"、“臺灣海膜"應為臺灣產杜氏海膜。分子親源關係分析也表明了臺灣本島與離島各地的羽狀分枝型海膜目前應該只有杜氏海膜與在臺數量較為稀少的湯都海膜兩種。杜氏海膜與湯都海膜其藻體外觀除了葉片與連接基部的短柄長短有相當程度的差異外,其餘外觀形態特徵則相當接近。杜氏海膜在臺灣會因地理環境不同使得外觀有所差異,使得杜氏海膜有多變的外觀,因此將杜氏海膜細分為四個地點探討(臺灣東北角、南部、離島蘭嶼、綠島);2)過去臺灣匍匐葉狀型海膜的紀錄只有“平展海膜",其過去紀錄中的描述與“葉狀海膜–1"相似,但利用分子序列與國外平展海膜比對,其結果尚未配對到相同或類似的藻種,如要認定為新種,則需未來需加入更多世界各地葉片型海膜藻才可確認。同為匍匐葉狀型海膜的“葉狀海膜–1"與“葉狀海膜–2"其藻體外觀有部分不同,“葉狀海膜–1"較大、質地柔軟黏滑、邊緣撕裂狀較多、某些藻體有孔洞與短柄,“葉狀海膜–2"則較小、質地韌似木耳、邊緣為無撕裂狀居多、無孔洞與短柄,為不同藻種;3)分子親源關係分析結果亦顯示,海膜藻屬演化群嵌入了其它屬藻種,為多源體系,有待來日以多基因序列分析再進一步探討。
Halymenia is one of the large marine red algal genera and is widely distributed in subtidal zones of the tropical, subtropical and temperate regions; it distribution widely in Taiwan and occurs throughout the year, but western coast is more scarce. The genus comprises about 69 species around the world. Based on the thallus morphology, this genus can be grouped into two types: Feathery-type and Prostrate -type. The generic morphology include mostly dark red, soft slimy, with a discoid holdfast. Previous studies on the taxonomy of Halymenia from Taiwan were mainly focused on morphological observations and cause to some taxonomic confusion and synonymy. In this study, the taxonomy of Halymenia is revised based on comparative anatomy, inter- specific relationships inferred using rbcL sequence analyses, and description of the auxiliary cell ampullae. The molecular analyses showed that there are four species identified: 1)Halymenia durvillei, a previously recorded species in Taiwan; 2)Halymenia tondoana, a new record in Taiwan; 3)Halymenia“dilatata-1"and 4) Halymenia“dilatata-2", two new species. The four Halymenia species are easily mistaken for known species, because these Halymenia have similar gross morphological, but the use of molecular and morphology characteristics can successfully distinguish these algal species.Base on the morphology and molecular phylogenetic analyse of the four Halymenia species:1)H.durvillei and H. tondoana both are the Feathery-type, with similar morphology features, but the blade and stipe are different. H.durvillei expressed diverse morphology due to environmental manual exerted in the geography of the habitat. In this study, we investigated Halymenia durvillei collection from four areas (Taiwan northeastern, southern, Orchid Island, Green Island). Molecular sequence analysis suggested that the H. ceylanica, H. formosa, H. floresia, H. microcarpa, previously recorded in Taiwan are H. durvillei; 2)Both H.“dilatata-1" and H.“dilatata-2"belong to the Prostrate-type, the two algal species are different in morphology, H.“dilatata-1" is bigger, soft slimy, margin lacerate, stipes more conspicuous than H.“dilatata-2". In the past, the occurring of the Prostrate-type only H.“dilatata"in Taiwan, and similar of describes with H.“dilatata-1", but can not found the same sequence when use molecular compare other country species, if want to make sure, we need to compare a lot more other place with Halymenia. 3)Molecular phylogenetic analysis of the study showed that Halymenia is polyphyletic. more genes are required to define the Halymenia group in the future.
目錄

謝辭 I
摘要 II
Abstract III
目錄 IV
圖目錄 VI
表目錄 VII
相關專有名詞解釋 VIII
第一章、前言 1
1.1 海膜藻屬簡介 1
1.2 海膜藻目海膜藻科分類研究歷史 2
1.2.1海膜藻目研究歷史 2
1.2.2海膜藻科研究歷史 3
1.2.3海膜藻科各屬分類情形 4
1.3 臺灣產海膜藻屬研究歷史及分類情況 6
1.4海膜藻屬藻種生活史及相關形態發育 7
1.5 研究動機與目的 9
第二章、材料與方法 11
2.1 藻種採集與保存 11
2.1.1 蠟液標本製作流程 11
2.2 分子序列分析方法 13
2.2.1 去氧核糖核酸萃取(DNA Extraction) 13
2.2.2基因擴增(Gene Amplification):聚合酶連鎖反應(Polymerase chain reaction;PCR) 14
2.2.3 定序反應及基因定序(Sequencing Reaction and Gene Sequencing) 14
2.2.4 親源關係分析(Phylogenetic Analysis) 15
2.3 形態觀察 16
2.3.1 染料配製 16
2.3.2 切片染色方法 17
第三章 結果 19
3.1 海膜藻屬與相關藻種親源關係樹分析 19
3.2藻體觀察結果 25
3.2.1 Halymenia durvillei 26
3.2.2 Halymenia tondoana 50
3.2.3 Halymenia“dilatata–1" 56
3.2.4 Halymenia“dilatata–2" 69
第四章、討論與結論 73
第五章、參考文獻 81

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