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題 名 | 分蔥潛隱病毒親緣分析及應用巢式聚合酶鏈鎖反應作分群偵測=Phylogenetic Analysis and Grouping Detection of Species Shallot latent virus by Nested-PCR |
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作 者 | 林玫珠; 林羿廷; 蔡錦慧; 鄧汀欽; | 書刊名 | 臺灣農業研究 |
卷 期 | 67:3 2018.09[民107.09] |
頁 次 | 頁270-282 |
分類號 | 433.4 |
關鍵詞 | 分蔥潛隱病毒; 蔥科作物; 親緣分析; 巢式-聚合酶鏈鎖反應; Shallot latent virus; Allium species; Phylogenetic analysis; Nested-polymerase chain reaction; |
語 文 | 中文(Chinese) |
中文摘要 | 搜尋美國國家生物技術資訊中心(National Center for Biotechnology Information; NCBI) GenBank登錄之分蔥潛隱病毒種(species Shallot latent virus; SLV)分離株鞘蛋白基因核苷酸序列,進行親緣分析。本研究可將現有SLV分為3個分類群:第一群(G1)、第二群(G2)、第三群(G3)。2015-2016年間採集台灣的蔥屬植物樣品,進行酶聯抗體免疫分析(enzyme-linked immunosorbent assay; ELISA)及反轉錄-聚合酶鏈鎖反應(reverse transcription-polymerase chain reaction; RT-PCR),檢出10株SLV。將其鞘蛋白基因核苷酸序列定序,其中6株SLV鞘蛋白基因核苷酸長度為894 bp,屬於第一群(G1);另外4株鞘蛋白基因核苷酸長度為885 bp,屬於第三群(G3)。不同SLV之病毒株鞘蛋白基因核苷酸序列的相同度大於74.3%,胺基酸序列相同度大於84.4%。根據所分類的各群病毒鞘蛋白基因核苷酸序列之差異,設計個別的專一性引子,進行RT-PCR,並配合巢式-聚合酶鏈鎖反應(nested-polymerase chain reaction; nested-PCR),可針對第一群(G1)病毒株增幅出187 bp的條帶,第二群(G2)病毒株增幅出548 bp的條帶,第三群(G3)病毒株增幅出540 bp的條帶。將134個經RTPCR測得SLV的蔥科作物樣本進行上述的nested-PCR,其中52個蒜球與25個蒜苗的病毒株中,分別有51個與24個病毒株為第一群;11個韭菜的病毒株皆為第三群,其中有9個另有檢出第一群者;34個青蔥的SLV中,有5株為第一群,19株為第三群、15株均有檢出第一群與第三群者。由本研究所檢測的台灣蔥科作物中,並無發現第二群的SLV存在。 |
英文摘要 | The coat protein (CP) gene nucleotide sequences of species Shallot latent virus (SLV) collected from National Center for Biotechnology Information (NCBI)-GenBank database were tested for phylogenetic analysis. All the current SLV isolates tested were clustered into three groups: G1, G2 and G3. In 2015-2016, a total of 10 SLV-infected Allium materials were collected from a survey of SLV occurred in Taiwan by detecting the virus with enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). The CP genes of each isolates were cloned and sequenced, among 10 isolates tested, 6 isolates were clustered into G1 with 894 bp nucleotides. The others belong to G3 with 885 bp nucleotides of CP gene. Based on divergence of CP nucleotide sequences, the group specific primer pairs were designed to differentiate the groups. Following by a RT-PCR, the nested-polymerase chain reaction (nested-PCR) was performed, and the amplified DNA was 187 bp, 548 bp and 540 bp for G1, G2 and G3, respectively. Accordingly, 134 SLV-infected samples were collected and detected by RT-PCR and nested-PCR as described above. There were 51 out of 52 garlic bulbs and 24 out of 25 garlic plants were identified as G1. Moreover, a total of 11 samples from Chinese leeks were defined as G3, and 9 samples out of them were complex with G1 and G3. The other 34 samples tested from green onions were consisted of 5 belonged G1, 19 belonged G3 and 15 were complex with G1 and G3. In this study, no SLV isolates belonged G2 were found from Allium in Taiwan. |
本系統中英文摘要資訊取自各篇刊載內容。