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題名 | 臺灣山藥上分離的蠶豆萎凋病毒2兩個株系之比較鑑定=Comparative Characterization of Two Isolates of Broad Bean Wilt Virus 2 Infecting Yams (Dioscorea spp.) in Taiwan |
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作者 | 鄧汀欽; 林羿廷; 蔡錦慧; 廖吉彥; 高瑞隆; | 書刊名 | 臺灣農業研究 |
卷期 | 63:4 2014.12[民103.12] |
頁次 | 頁282-290 |
分類號 | 434.3 |
關鍵詞 | 豆類病毒屬; 引子對; 反轉錄聚合酶鏈式反應; 核苷酸序列; 親緣分析; Fabavirus; Primer; RT-PCR; Nucleotide sequence; Phylogenetic analysis; |
語文 | 中文(Chinese) |
中文摘要 | 採集畸形嵌紋的「台農1號」山藥病株及黃化嵌紋病徵的日本山藥病株,接種奎藜經單斑分離後分別得到病毒分離株“YA3"與“Yam-7"。陰染法電子顯微鏡觀察,均為球形病毒。兩者都可被蠶豆萎凋病毒2(Broadbean wilt virus 2; BBWV-2)抗體檢出。接種後潛伏感染山藥品種(系),翌年才在山藥新葉出現病徵且可檢測到病原。以豆類病毒屬(Fabavirus)廣效性引子對Fab5'R1F/Fab5'R1R進行反轉錄–聚合酶鏈鎖反應(RT-PCR),兩者都增幅得到一390 bp的DNA產物,此段核苷酸序列YA3與Yam-7相同度為94.1%,與中國北柴胡BBWV-2分離株相同度為93.1%。再以BBWV-2專一性與引子對BBWVSSP/BBWVKMRM進行RT-PCR,YA3與Yam-7各增幅得一320 bp的DNA產物,此段核苷酸序列YA3與Yam-7的相同度為88.1%,與日本的長形山藥BBWV-2分離株相同度為86.3%,但Yam-7與此日本分離株相同度為96.5%。親緣分析圖也顯示YA3與Yam-7兩株病毒都屬於一種BBWV-2,而異於其他同屬的病毒種。這是BBWV-2在台灣感染山藥的首發報告。 |
英文摘要 | Leaf samples with mosaic and malformation on "Dioscorea alata" and that with yellowing and mosaic symptoms on "Diosocoreab atatas" were collected for inoculation tests. Both of them developed local lesions on "Chenopodium quinoa", and two virus isolates 'YA3' and 'Yam-7' were obtained through single lesion isolation. YA3 and Yam-7 both with spherical visions were observed by electron microscope and they reacted with antibody to "Broad bean wilt virus" 2 (BBWV-2) were detected by ELISA. When back-inoculated to "Dioscorea" spp., tested plants of some cultigens with latent infections showed symptoms only in the following years and the virus was detected by ELISA. With "Fabavirus" common primers Fab5'R1F/Fab5'R1R, reverse transcription-polymerase chain reaction (RTPCR) was conducted to amplify a 390 bp DNA product from both of YA3 and Yam-7. The nucleotide sequence of this fragment sharing 94.1% identities between YA3 and Yam-7. Comparing with other isolates, YA3 shared 93.1% identities with "Bupleurum chinense" isolate of BBWV-2. Moreover, with BBWV-2 specific primers BBWVSSP/BBWVKMRM to conduct RT-PCR, both YA3 and Yam-7 obtained a 320 bp product. The identities between YA3 and Yam-7 of this nucleotide sequence fragment were only 88.1%. Comparing with other isolates, YA3 was 86.3% identical to the Japanese isolate of BBWV-2 infecting yams. However, this Japanese isolate shared 96.5% identities with yam-7. Results of phylogenetic analysis also show that YA3 and Yam-7 are closely related and belonged to a BBWV- 2, which is different from other species of fabaviruses. This is the first report of BBWV-2 infecting yams in Taiwan. |
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