青春激素誘發德國蜚蠊雌蟲基因表現之分析

Analysis of genes upregulated by juvenile hormone in the female German cockroach, Blattella germanica (L.) (Dictyoptera: Blattellidae)

陸子鈞(Tzu-Chun Lu)

指導教授 ： 李後晶

國立臺灣大學/生物資源暨農學院/昆蟲學研究所/碩士(2013年)

https://doi.org/10.6342/NTU.2013.02278

摘要

德國蜚蠊（Blattella germanica）雌蟲的發育、生殖及行為受到體內青春激素濃度的調控。本研究即利用青春激素類似物－烯蟲酯（methoprene）施用於三日齡的雌成蟲，並藉由抑制消除雜合技術（Suppression Subtractive Hybridization, SSH），找出施用烯蟲酯後特殊表現的基因。雜合後的cDNA資料庫，包含20個特殊表現的基因片段，這些基因潛在受到青春激素濃度調控。與線上資料庫比對的結果，其中9個基因片段，其中功能各涉及卵巢發育、能量補給、蛋白產物合成、細胞分裂、解毒等，另外有11個基因則功能未知。功能已知的16號基因，和能量調控有關，但尚未有更詳細的研究；再加上三個功能未知的基因，共四個基因，由進一步研究得到更多資訊。藉由即時聚合酶連鎖反應（real-time PCR）技術，確認這四個基因受到青春激素誘發表現，也顯示在不同日齡的雌蟲間有不同的表現量；此外，在不同器官也有不同表現量，暗示著可能各有不同功能。由於過去研究發現，青春激素濃度會間接調控德國蜚蠊雌蟲的日週律動以及卵巢發育，因此我們也藉由RNAi技術，測試這四個基因是否屬於調控日週律動或卵巢發育的調控因子。結果顯示，注射雙股RNA後，四個基因的表現量降低，其中三個基因BGME02、BGME05及BGME16的表現量被抑制之後，卵巢的發育減緩，但日週律動卻無明顯差異。因此我們認為這三個基因是青春激素下游調控卵巢發育的基因，但不直接調控雌蟲的日週律動。

關鍵字

德國蜚蠊 ； 青春激素 ； 日週律動 ； RNAi ； 抑制消除雜合法

並列摘要

Juvenile hormone (JH) is the major hormone controlling the development, behavior and reproductive cycles of the female German cockroach, Blattella germanica (L.) (Dictyoptera: Blattellidae). In this study, JH analog - methoprene was topically applied on 3-day old females before with Suppression Subtractive Hybridization (SSH) to identify the genes that were particularly expressed. The SSH cDNA library composed with 20 distinctive sequences of genes. Nine of those genes were involved in ovarial development, energy supply, protein synthesis, cell division or detoxification, and rest of genes possessed unknown functions. For functional studies, RNAi was used to knock-down the expression level of four genes, and dsBGME02, dsBGME05 and dsBGME16 injection inhibit ovary development, but RNAi of all four genes do not affect the locomotor rhythmicity. Based on these findings, we found three genes, which are BGME02, BGME05 and BGME16 mediated by JH downstream regulation and ovary development, but do not participated in control of locomotion.

並列關鍵字

Blattella germanica ； Circadian rhythm ； Juvenile Hormone (JH) ； RNAi ； Suppression Subtractive Hybridization

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