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研究生:陳怡華
研究生(外文):Chen, Yi-Hua
論文名稱:吳郭魚對於低溫適應之研究
論文名稱(外文):A study of cold acclimation of Tilapia(Oreochromis niloticus)
指導教授:陸振岡
指導教授(外文):Dr.Jenn-Kan Lu
口試委員:陳鴻鳴秦宗顯陸振岡
口試委員(外文):Dr.Hong-Ming ChenChin, Tzung-ShianDr.Jenn-Kan Lu
口試日期:2015-07-27
學位類別:碩士
校院名稱:國立臺灣海洋大學
系所名稱:水產養殖學系
學門:農業科學學門
學類:漁業學類
論文種類:學術論文
論文出版年:2015
畢業學年度:103
語文別:中文
論文頁數:71
中文關鍵詞:低溫吳郭魚生理乙醯膽鹼
外文關鍵詞:low-temperatureTilapiaphysiologicalacetylcholine
相關次數:
  • 被引用被引用:3
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  • 下載下載:48
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吳郭魚是台灣淡水主要養殖魚類之一,也是世界性養殖魚種,更是未來人類動物性蛋白質主要來源之一。養殖吳郭魚有很高的經濟利益,因為其天然抗病性強,雜食性魚種,繁殖能力強,幼魚存活率高,養殖所需空間及其小,再加上吳郭魚成長快速,6個月即可達於上市之體型。目前台灣養殖吳郭魚主要分布於中南部,冬季寒流來襲時會使得水溫過低,導致低溫下的細菌性感染增加,引發疾病導致死亡。本實驗研究目的為利用低溫刺激對吳郭魚之影響進行分析以作為分子育種的基礎,達到能夠耐受溫度逆境環境的新品系吳郭魚。本研究實驗方法是將吳郭魚在26℃下馴養3周後,分別放入10℃、15℃、20℃、26℃(對照組)的水環境中。在不同溫度處理2hr、6hr、12hr、18hr後進行抽血及採樣,對各組魚樣品進行血清生化指數酶活力測定。發現丙酮酸激酶在低溫下和對照組有顯著差異(P<0.05);低溫下ATP含量較對照組來的高; 而神經傳導物質乙醯膽鹼和乙醯膽鹼脂酶測定則有顯著差異(P<0.05)。最後利用其他物種已知的低溫相關基因來篩選主要的目標基因,並進行克隆和表達分析。目標基因有5個分別為冷誘導RNA結合蛋白(cold-inducible RNA-binding protein,CIRP)、高速遷移蛋白(high mobility group protein B1,HMGB1)、載脂蛋白(Apolipoprtein,APO-AⅣ)、乙醯膽鹼脂酶(Acetylcholinesterase,AchE),由Q-PCR定量圖譜顯示:CIRP和HMGB1在腦組織、鰓組織、心臟、肝臟、肌肉等均有表達,但是HMGB1基因在鰓組織中表達較弱無顯著(P>0.05),而APO-AIV基因在腦組織中有明顯的差異性(P<0.05),鰓組織則表達較低。
Tilapia is one of Taiwan's major freshwater fish, is also the world aquaculture species, is one of the main source of future human animal protein. Aquaculture tilapia has high economic benefit, because of its strong natural resistance to disease, reproductive ability, juvenile fish survival rate is high, small breeding space, coupled with the rapid growth of tilapia, 6 months reach listed on the body type. At present, Taiwan aquaculture tilapia are mainly distributed in South Central, in the hot summer makes the water temperature is too high, causing disease and death, like streptococcus. Winter cold will make the water temperature is too low, cause bacterial infection increased under low temperature. This study is the Tilapia at 26 ℃ training support three weeks later, were placed in 10 ℃, 15 ℃, 20 ℃, 26 ℃ (control) in the aquatic environment. Treated at different temperatures 2hr, 6hr, 12hr, 18hr and for blood sampling, sample each group of fish serum biochemical indices enzyme activity assay, pyruvate kinase found significant differences at low temperatures and in the control (P<0.05); ATP content at relatively low temperatures in the control to high; and neurotransmitter acetylcholine and acetyl cholinesterase, there are significant differences measured (P<0.05). And the use of technology as a paraffin-embedded tissue sections on the organization can be directly observed. At last, other species of cold-related genes known to filter the main objective gene, and cloning and expression analysis. There are five gene were cold-inducible RNA binding protein (cold-inducible RNA-binding protein, CIRP), high-speed migration protein 1 (high mobility group protein B1, HMGB1), Apolipoprotein (Apolipoprtein, APO-AⅣ), ethyl acyl cholinesterase (Acetylcholinesterase, AchE), acetylcholine (Acetylcholine, Ach), the Q-PCR quantitative map display: CIRP and HMGB1 were expressed in brain tissue, gill tissue, heart, liver, muscle,but HMGB1 gene expression in gill tissue is weak(P>0.05), and APO-AIV gene have significant differences(P<0.05).
謝辭 i
中文摘要 ii
Abstract iii
目錄 iv
表目錄 vii
圖目錄 viii
壹、前言 8
一、溫度影響魚類生理性狀 8
二、魚類抗寒相關研究進展 10
三、本研究之目標 18
貳、材料與方法 19
一、實驗設計 19
二、藥品及儀器 19
三、實驗動物 19
四、實驗方法 19
肆、結果 34
一、低溫下吳郭魚丙酮酸激酶活性測定 34
二、低溫下吳郭魚胰蛋白酶活性測定 34
三、低溫下吳郭魚ATP含量測定 34
四、低溫下吳郭魚乙醯膽鹼含量測定 34
五、低溫下吳郭魚乙醯膽鹼酯酶活性測定 35
六、低溫下目標基因在吳郭魚之各組織之間的表現分布情形 35
七、低溫下吳郭魚組織學的變化 36
伍、討論 38
一、低溫脅迫下吳郭魚行為能力的變化 38
二、低溫下吳郭魚血清生化指數酶活力測定 38
三、低溫下目標基因在吳郭魚之各組織之間的表現分布情形 39
四、低溫下吳郭魚組織學的變化 41
陸、參考文獻 42
柒、圖表 48
附錄一 66
附錄二 68


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