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標題: 新德里番茄捲葉病毒寄主適應特性關鍵因子之研究
Characterization of the critical factors for host adaptation of Tomato leaf curl New Delhi virus
作者: Yao-Wei Kuo
關鍵字: 新德里番茄捲葉病毒
Tomato leaf curl New Delhi virus
host adaptability
movement protein
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摘要: 雙生病毒科 (Geminiviridae) 的病毒遍及全世界,並且寄主範圍十分廣泛,對於現今全球的農業經濟危害十分嚴重。新德里番茄捲葉病毒 (Tomato leaf curl New Delhi virus, ToLCNDV) 屬於雙生病毒科豆類金黃嵌紋病毒屬 (Begomovirus),常被報導於熱帶與亞熱帶地區,能感染許多種植物,其中又以葫蘆科與茄科的作物最為嚴重,例如印度與巴基斯坦的番茄,以及泰國的葫蘆科作物等。ToLCNDV具有二條約2.7 kb的單股環狀DNA,分別命名為DNA-A和DNA-B。DNA-A內編碼著鞘蛋白、複製與基因表現相關的蛋白;而DNA-B編碼的蛋白主要與病毒移動相關。於2007年四月,本實驗室從台灣的東方型甜瓜銀輝栽培種 (Cucumis melo var. makuwa cv. Sliver Light) 上分離出一種能機械接種的ToLCNDV分離株,將其命名為新德里番茄捲葉病毒東方甜瓜分離株 (ToLCNDV oriental melon isolate, ToLCNDV-OM),ToLCNDV-OM可感染菸草及番茄兩種寄主;另一分離株則是從泰國的胡瓜 (C. sativus L.) 上所分離出來,將其命名為新德里番茄捲葉病毒黃瓜分離株 (ToLCNDV cucumber isolate, ToLCNDV-CB),而此分離株無法感染番茄。因此本實驗目的是利用基因重組的策略,來探討影響此兩種分離株對於番茄寄主適應差異之關鍵因子為何。將各種不同基因置換的具感染力病毒構築,分別以農桿菌注射接種於圓葉菸草 (Nicotiana benthamiana) 以及番茄農友301栽培種 (Lycopersicon esculentum Mill var. Farmers 301),其接種結果發現ToLCNDV-OM的移動蛋白 (movement protein, MP) 是影響病毒感染番茄的關鍵蛋白。而又藉由胺基酸點突變的方式進一步發現,移動蛋白的第19個胺基酸 (谷胺酸,E19) 是影響ToLCNDV-OM對於番茄寄主適應性的關鍵因子。而ToLCNDV- CB除了移動蛋白的第19個胺基酸,第3個胺基酸 (異白胺酸,I3) 也會影響其對番茄的寄主適應性。本研究藉由同一種病毒的兩種分離株,利用基因重組和點突變的策略,精確地指出移動蛋白為影響ToLCNDV寄主適應性的關鍵因子。
Members in the Geminiviridae have wide host range including economically important crops and are cosmopolitan in distribution. Tomato leaf curl New Delhi virus (ToLCNDV), a member of genus Begomovirus, infects a wide range of plants in tropical and subtropical regions, especially cucurbitaceous and solanaceous crops. It was isolated from tomato in India and Pakistan and from cucurbitaceous plants in Thailand. ToLCNDV contains bipartite genome, designated as DNA-A and DNA-B and are approximately 2.7 kb each in size. DNA-A encodes coat protein, replication-associated proteins and proteins involved in gene expression. Similarly, DNA-B encodes proteins responsible for viral movement. In April 2007, a mechanically transmissible isolate of ToLCNDV which was later designated as ToLCNDV oriental melon (ToLCNDV-OM) was isolated from oriental melon (C. melo var. makuwa cv. Sliver Light) in Taiwan. This ToLCNDV-OM isolate can also infect tomato (L. esculentum Mill. var. Farmers 301). However, another isolate designated as ToLCNDV cucumber (ToLCNDV-CB) and was originally isolated from cucumber can not infect tomato. In order to investigate the factors that affect its host adaptability, gene recombination between ToLCNDV-OM and ToLCNDV-CB was done. Tobacco (N. benthamiana) and tomato plants grown in green house were challenged with the recombinant viruses via agroinfiltration. The inoculation results showed that the exchange of the gene coding for movement protein (MP) affected the virus infectivity on tomato. Furthermore, we have also identified that the 19th amino acid residue (glutamate, E19) of ToLCNDV-OM MP gene determines the tomato adaptation via the protein point mutation protocol. It was also shown that not only19th amino acid residue of ToLCNDV-CB MP gene, but the 3rd amino acid residue (isoleucine, I3) also has influence in its adaptability in the tomato. In this study, we used two isolates of one virus to prove that the MP of ToLCNDV is the critical factor for its host adaptability via gene recombination and point mutation.
文章公開時間: 10000-01-01
Appears in Collections:植物病理學系



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