臺灣博碩士論文加值系統

海洋腔腸動物與蟲黃藻以胞內共生相互形成美麗的珊瑚礁生態。蟲黃藻以胞內共生的方式，生活在腔腸動物宿主消化腔內胚層的細胞內。傳統認知，腔腸動物共生小體內腔pH值為6~7，而最新研究指出可能為pH 4以下。本實驗藉由比較不同宿主體內共生小體的差異，及蟲黃藻在不同pH值生長的變化，找尋出最適不同種類蟲黃藻生長的環境。本實驗共測試八種腔腸動物及一種二枚貝生物，結果指出Aiptasia pulchella、Bolocera mcmurrichi、Fungiidae spp.及Sinularia polydactyla等腔腸動物共生的蟲黃藻，在我們所提供的培養基環境下，皆有其適合生長的環境範圍。免疫螢光染色發現，其共生小體上帶有V-ATPase D蛋白的族群約為10~20%左右；宿主細胞酸性定量分析結果也與免疫螢光染色結果相符，推測其共生小體內腔值應在中性到弱酸性之間，而在不同種類間也有差異性。

關鍵字：腔腸動物、共生小體、蟲黃藻、pH值

The cnidarians form the amazing coral reef ecosystem mutually with zooxanthellae, its foundation was based on the intracellular symbiosis. By endosymbiosis, zooxanthellae were capable of living in cnidarian host gastrodermal endoderm cells with large numbers. Traditionally, the pH of symbiosomal lumen was thought to be 6-7. However, a recently report by utilizing pH indicators indicated that the pH of symbiosomal lumen in coral cells is below 4, revealing that the regulatory mechanism of symbiosome acidification should be clarified. Therefore, this study hypothesized that the pH of symbiosomal lumen in different cnidarian were different. Through culturing the fresh-isolated zooxanthellae with the different medium, the most suitable pH range of different zooxanthellae were evaluated. In this study, a total of eight of cnidarians species, and one test bivalves. The current results indicate that the zooxanthellae from Aiptasia pulchella, Bolocera mcmurrichi, Fungiidae spp. and Sinularia polydactyla, which were can growth at 6.2 and 8.6. The immunofluorescence showed that V-ATPase D was not abundantly distributed on the symbiosomal membrane of coral cells, suggesting symbiosomes were not acidic vesicles.

Keyword：Cnidarians、Symbiosome、Zooxanthellae、pH

碩博士論文授權書 I
學位論文考試審定書 II
碩士論文著作權歸屬協議書 III
摘要 IV
Abstract V
誌謝 VI
目錄 VIII
表目錄 X
圖目錄 XI
壹、前言 1
一、共生的定義 1
二、胞內共生對腔腸動物的意義 1
三、蟲黃藻的分類與型態 2
四、蟲黃藻與硨磲貝的相互性 3
五、胞內共生的建立 4
六、胞噬小體的形成與酸化 5
七、蟲黃藻與腔腸動物的關係 7
貳、研究目的 9
參、材料與方法 10
一、實驗架構 10
二、實驗生物取得及飼養 11
三、實驗器材及藥品 13
四、蟲黃藻取得及培養 16
五、共生態蟲黃藻於不同pH值海水下的生長變化 16
六、共生態蟲黃藻於不同pH值培養基下的生長變化 17
七、抗體(antibodies) 18
八、西方墨點法(western blot) 18
九、免疫螢光染色(immunofluorescence) 19
十、細胞內酸性囊泡測定 20
肆、結果 22
一、共生態蟲黃藻於不同pH值海水下的生長變化 22
二、共生態蟲黃藻於不同pH值培養基下的生長變化 26
三、V-ATPase D抗體的專一性 29
四、V-ATPase 蛋白在共生小體上的分佈 29
五、宿主共生小體酸性定量分析結果 30
伍、討論 31
一、共生態蟲黃藻於不同pH值海水下的生長變化 31
二、共生態蟲黃藻於不同pH值培養基下的生長變化 33
三、V-ATPase D抗體的專一性確認及使用 36
四、宿主共生小體酸性定量分析 36
陸、結論 39
柒、圖表 40
捌、參考文獻 66
玖、附錄 73

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