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研究生:古金台
研究生(外文):Chin-Tai Ku
論文名稱:台灣小菜蛾微粒子蟲分離株(Nosemasp.PX1&2)之特性研究
論文名稱(外文):The characteristics of two Nosema isolates (Nosema sp. PX1 & 2) from Plutella xylostella in Taiwan
指導教授:王重雄
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:昆蟲學研究所
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:62
中文關鍵詞:小菜蛾微孢子蟲
外文關鍵詞:microsporidium
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由實驗室飼養族群分離之兩株小菜蛾微粒子蟲分離株,此兩株微粒子蟲並非是已報導的小菜蛾變形微孢子蟲 (Vairimorpha sp.)。經西方轉印法、家蠶微粒子蟲 (Nosema bombycis) 之特異性引子檢測,確定此兩株應屬於不同分離株,分別命名小菜蛾微粒子蟲分離株1和分離株2 (Nosema sp. PX 1 & Nosema sp. PX 2)。以電子顯微鏡觀察PX 1和PX 2之生活史及超微構造,二者在超微構造上的特徴並無明顯差異。孢子呈長橢圓形,表面光滑,大小約3.93 μm × 1.38 μm,只有一種型式大小的孢子,並未發現如Vairimorpha屬之大小孢子及八倍體孢子。此小菜蛾分離株具有典型的Nosema屬之生活史特徵,如母孢子、孢子母細胞及孢子之各個時期,皆具有雙倍體核並以二分裂法繁殖。母孢子具有電子緻密的細胞膜,在細胞內部有時會出現延長的空泡。孢子母細胞為電子緻密的細胞膜。孢子具有內外孢子壁 (外孢子壁的厚度為9.09 nm,內孢子壁為45.45 nm)、鰓葉狀的極囊、極絲 (12-13圈)、雙核及後囊。探討本研究之二小菜蛾分離株與已發表模式種N. bombycis及近緣種N. spodopterae之核糖體RNA (ribosomal RNA)、alpha-tubulin、beta-tubulin及RPB1(DNA dependent RNA polymeraseⅡ largest subunit) 之序列比對和親緣關係,發現此四株微孢子蟲在這些基因的序列相似度非常高 (高於 95%)。且在rRNA基因的排列是相似,由5’至3’為LSU-rRNA-ITS-
SSUrRNA。通常較少利用非small subunit rRNA序列來做親緣關係的分析,但我們藉由上述基因所構築的親緣關係樹,可推論出4株微孢子蟲是親緣關係很近的微粒子蟲,但PX1與N. bombycis及N. spodopterae關係更近。經由西方轉印法可知此兩者的孢子可溶性蛋白質具有不同的圖譜,再利用二維式電泳將此二小菜蛾分離株及N. spodopterae之可溶性蛋白質解析,PX 2顯現較不同的圖譜,PX 1則與N. spodopterae較相似,顯示二維蛋白質圖譜在同屬間常具有相同的型式。綜合以上的結果,PX之二分離株是屬微粒子屬,與N. bombycis和N. spodopterae親緣關係很接近,但PX 2分離株呈現較多不同的各種生化與分子上的特徴。這些證據證實PX 2可能為一個新種而PX 1則是新的亞種。
目錄
中文摘要……………………………………………………………………… 1
英文摘要…………..………………………………………………………….. 3
壹、前言………..………………………………………………………….. 5
貳、前人研究……………………………………………………………….. 7
參、材料與方法…………………………………………………………….. 13
肆、結果…………………………………………………………………….. 18
伍、討論…………………………………………………………………….. 23
陸、參考文獻……………………………………………………………….. 32
柒、圖表…………………………………………………………………….. 38
附錄一.……………………………………………………………………….. 60
致謝…………………………………………………………………………… 62
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