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| 題 名 | 感染沙漠玫瑰的緬梔嵌紋病毒(Plumeria mosaic virus)之鑑定與檢測=Identification and Detection of Plumeria mosaic virus Infecting Desert Rose (Adenium obesum) |
|---|---|
| 作 者 | 林羿廷; 蔡錦慧; 林玫珠; 鄧汀欽; | 書刊名 | 臺灣農業研究 |
| 卷 期 | 69:1 2020.03[民109.03] |
| 頁 次 | 頁77-89 |
| 分類號 | 435.415 |
| 關鍵詞 | 臺灣; 分離株; 菸草嵌紋病毒屬; 雞蛋花嵌紋病毒; Taiwan; Isolate; Tobamovirus; Frangipani mosaic virus; |
| 語 文 | 中文(Chinese) |
| 中文摘要 | 2013年高雄「大紅花」沙漠玫瑰,疑似病毒感染,經單斑接種於奎藜(Chenopodium quinoa Willd.),分離得到一個病毒純系的分離株DR-TW。經免疫分析顯示此分離株與雞蛋花嵌紋病毒(Frangipani mosaic virus; FrMV)及番茄嵌紋病毒 (Tomato mosaic virus; ToMV)等菸草嵌紋病毒屬(Tobamovirus)的病毒抗體有交互反應。汁液接種番杏、紅藜(Chenopodium amaranticolor Coste et Reyn)、奎藜、綠藜(Chenopodium murale L.)、千日紅(Gomphrena globosa L.)及曼陀羅後都呈局部病斑,菸草、雞蛋花及沙漠玫瑰植株則形成系統性嵌紋,但對辣椒、番茄等茄科植物的病原性及致病力均弱於ToMV。以菸草嵌紋病毒屬廣效性引子對TobRT up1/TobRT do2進行反轉錄-聚合酶連鎖反應(reverse transcription-polymerase chain reaction; RT-PCR),增幅出568bp的核酸片段。經定序後與GenBank中的核苷酸序列進行比對,結果顯示沙漠玫瑰台灣分離株(DR-TW)與緬梔嵌紋病毒(Plumeria mosaic virus; PluMV)的相同度高達98%。進行DR-TW基因體定序,得核苷酸全長6,683 bp(GenBank accession number KX881422),共轉錄出188.3 kDa及130.4 kDa的複製酶、28.7 kDa的移動蛋白及19.0 kDa的鞘蛋白。設計PluMV專一性引子對(PLuMVcp-up/PLuMVcp-dw),經RT-PCR可增幅出PluMV的核酸片段643 bp,其他8個供試菸草嵌紋病毒屬病毒則無。配合FrMV專一性引子對FrMVcp-up/FrMVcp-dw,以RT-PCR即可鑑別FrMV及PluMV。 |
| 英文摘要 | A diseased sample of desert rose (Adenium obesum) cv. Dahuahua showing virus-like symptom was found in Kaohsiung in 2013. The pure virus isolate (DR-TW) was obtained by inoculation to Chenopodium quinoa through single lesion transfer. Immunoassay showed that the isolate interacted crossly with the other tobamoviruses, including Frangipani mosaic virus (FrMV), Tomato mosaic virus (ToMV), etc. After sap inoculation, local lesions were developed on leaves of Tetragonia tetragonioides, Chenopodium amaranticolor, Chenopodium quinoa, Chenopodium murale, Gomphrena globosa and Datura stramonium; and systematic mosaic was showed on plants of Nicotiana benthamiana, frangipani and desert rose. However, the pathogenicity and virulence of DR-TW was milder than ToMV to tomatoes, peppers and eggplants. A pair of tobamovirus broad-spectrum primers (TobRT up1/TobRT do2) were applied in RT-PCR to amplify a nucleic acid fragment with 568 bp. Sequence alignment revealed that the RT-PCR fragment shared more than 98% identity with Plumeria mosaic virus (PluMV) but only 73% with FrMV. Another primers were designed and used for analysis of complete genome. The full length of genomic nucleotides with 6,683 bp was determined and deposited in GenBank under accession number KX881422. A total of 188.3 kDa and 130.4 kDa of replicates, 28.7 kDa of movement protein and 19.0 kDa of coat protein, were transcribed. A pair of PluMV-specific primers (PLuMVcp-up/PLuMVcp-dw) were designed to amplify a fragment of 643 bp nucleotides by RT-PCR, by which PluMV was detectable and could be distinguished from other tobamoviruses. The FrMV specific primers (FrMVcp-up/FrMVcp-dw) were also applied to differentiate the FrMV from PluMV by RT-PCR. |
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