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頁籤選單縮合
題 名 | 番椒斑駁病毒之鑑定與發生調查=Identification and Occurrence of Pepper mottle virus Isolated from Peppers in Taiwan |
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作 者 | 鄭櫻慧; 鄧汀欽; 陳金枝; 關政平; 張清安; | 書刊名 | 臺灣農業研究 |
卷 期 | 62:4 2013.12[民102.12] |
頁 次 | 頁360-371 |
分類號 | 435.27 |
關鍵詞 | 番椒斑駁病毒; 鞘蛋白; 抗血清; Pepper mottle virus; Coat protein; Antiserum; |
語 文 | 中文(Chinese) |
中文摘要 | 甜椒(Capsicum annuum)葉片出現嵌紋及扭曲變形疑似病毒感染的病徵,經直接法ELISA檢測,這些樣本可與辣椒葉脈斑駁病毒(Chilli veinal mottle viru;ChiVMV)的抗體反應,且汁液接種可感染Nicotiana benthamiana,但N. benthamiana中的病毒卻不與ChiVMV的抗體反應,也不感染紅藜、奎藜。經汁液系列稀釋後接種,可分離得一病毒純系,將其接種保存於N. benthamiana上,經寄主範圍試驗,結果只感染茄科植物,回接甜椒即在葉片出現嵌紋及扭曲變形病徵。抽取接種的N. benthamiana葉片之總量RNA,利用potyvirus的簡併式引子(HRP5/dT)進行反轉錄聚合酵素連鎖反應,可增幅得到1.3 kb的DNA片段。進行選殖及定序分析並與GenBank之核苷酸序列比對,與番椒斑駁病毒(Pepper mottle virus;PepMoV)VB分離株的鞘蛋白基因(Access No. AB126033)相同度達100%,因此鑑定本病毒是一株PepMoV。純化上述接種繁殖於N. benthamiana 的病毒分離株(PepMoV-TW1)製備多元抗血清,利用其抗體以ELISA進行病毒偵測,自2007至2011年採自台灣各地969個甜椒樣本中有20%可測到PepMoV,且多數與ChiVMV或番椒葉脈斑駁病毒(Pepper veinal mottle virus;PVMV)複合感染。因PepMoV與馬鈴薯病毒Y(Potato virus Y;PVY)、PVMV及ChiVMV等病毒有血清關係且彼此產生交互反應,使以往ELISA的結果之判別及病毒種類鑑定受到干擾,且甜椒中病毒複合感染的情況極為普遍,因此推測PepMoV可能之前就已存在於台灣。 |
英文摘要 | Bell pepper (Capsicum annuum L.) plants exhibiting systemic leaf mosaic and malformation were collected. By DAS-ELISA, these samples were positively reacted with antibody against Chilli veinal mottle virus (ChiVMV). After mechanical inoculation with sap extracted from these samples, unexpected mosaic symptoms were developed on upper leaves of Nicotiana benthamiana. The virus infecting N. benthamiana was non-reacted with antibody to ChiVMV when tested by DAS-ELISA. And the virus was non-infectious to Chenopodium amaranticolor and Chenopodium quinoa tested by sap-inoculation. Pure virus isolates were obtained from infected plants of N. benthamiana by mechanical inoculation with serially diluted saps of infected plants. Not only pepper plants were developed the mosaic and leaf distortion symptoms, but also some species of Solanaceae plants were found to be infected by this virus in the host range test. The total RNA extracted from leaves of inoculated N. benthamiana was subjected to RT-PCR. The potyvirus degenerate primers (Hrp5/Pot1) were used to amplify a RT-PCR product of 1.3 kb DNA fragment, which was then cloned and sequenced. Results of nucleotide sequencing revealed that the CP gene of the isolate was 100% identical to Pepper mottle virus (PepMoV)-VB (Access No. AB126033) and was concluded as an isolate of PepMoV. Polyclonal antiserum against PepMoV Taiwan isolate (PepMoV-TW1) was prepared by immunizing rabbit with purified virus that has been propagated in N. benthamiana. Use of its antibody in ELISA for virus detection, a total of 969 sweet pepper samples were collected from different areas of Taiwan and tested in 2007-2011. About 20% of all the survey samples were identified as PepMoV infection and most of them were complexly infected with ChiVMV, or Pepper veinal mottle virus (PVMV). It is speculated that PepMoV might already exist in Taiwan earlier, but it was ignored because the closely serological relationships among PepMoV, Potato virus Y (PVY), PVMV and ChiVMV could form cross-reactions with each other and that interfered with the judgment of ELISA. Besides, the virus complex infections were very common in plants of sweet pepper. |
本系統中英文摘要資訊取自各篇刊載內容。