竹嵌紋病毒衛星核酸在植物體之長距離移動

Long-distance trafficking of Bamboo mosaic virus satellite RNA in plant

10.6342/NTU201700078

張志豪

竹嵌紋病毒衛星核酸 ； 長距離移動 ； fibrillarin ； 核糖核蛋白複合體 ； Bamboo mosaic virus satellite RNA ； long-distance trafficking ； fibrillarin ； ribonucleoprotein complex

臺灣大學植物科學研究所學位論文

2017年

博士

林納生

英文

RNA的移動在調節植物生長、發育、營養運送和適應環境逆境中扮演相當重要的角色。對於RNA病毒而言，它的擴散需要藉由原生質聯絡絲進行細胞與細胞間移動，再靠維管束系統進行長距離移動。竹嵌紋病毒 (BaMV) 為單股RNA病毒，可表現複製酶、移動蛋白及鞘蛋白。竹嵌紋病毒衛星核酸(satBaMV)為BaMV的寄生物，需要病毒才能進行複製及包被，而satBaMV在植物體如何靠BaMV有效移動所知有限。 探討BaMV和satBaMV在植物體上的移動方向和速率，發現BaMV和satBaMV均先往植物下位葉系統性移動，再往植物上位葉移動進而感染整株植物，而BaMV在植物體的移動速率比satBaMV快。此外也發現，單獨接種satBaMV可能在植物體中長距離移動。為了確定satBaMV可單獨在植物體中長距離移動，利用嫁接實驗、北方墨點法和原位反轉錄聚合酶鏈式反應，發現轉殖株表達的satBaMV能單獨移動到嫁接的野生株植物接穗中，且satBaMV的N端序列決定其長距離移動能力。 前人研究指出satBaMV製造出P20蛋白質可能與satBaMV在植物體之長距離移動有關，本研究證實以維管束中表達P20的轉殖株能夠幫助缺少P20之satBaMV進行長距離移動。藉由嫁接實驗、免疫共沉澱和質譜分析發現P20複合體中包含核仁蛋白fibrillarin。除此之外，凝膠阻滯分析發現大腸桿菌表達fibrillarin能結合satBaMV。以嫁接或共同接種的實驗進一步發現fibrillarin基因靜默會抑制satBaMV的長距離移動，但對BaMV之移動則沒有影響。當BaMV和satBaMV共同感染時，satBaMV 核糖核蛋白複合體包含BaMV的移動蛋白、鞘蛋白、fibrillarin、BaMV RNA及satBaMV RNA，利用表現BaMV移動蛋白質之轉殖株亦證實BaMV移動蛋白能幫助satBaMV的長距離移動。本研究提供satBaMV蛋白複合體在植物中進行長距離移動與寄主病毒和衛星核酸之分子交互機制。

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