中文摘要：从南韩感染病毒的辣椒'Manidda'（P0抗性品种）叶片中分离出黄瓜花叶病毒Ca-P1菌株，即一种P0抗性断裂病毒。自从2006年首次报道CMV-Ca-P1后，这种可对辣椒生产造成损害的病毒在南韩经常被检测到。本研究构建了三种基于抵抗病毒侵染的防御系统转录后基因沉默的CMV RNAi载体。设计这些独立的载体（hpCMV1、hpCMV2和 hpCMV2 + 1 RNAi载体）生产CMV-Ca-P1 RNA-1（复制酶基因）的dsRNAs（dsRNAs含有每个hpCMV1 (1270-1629 nt)、hpCMV2 (1-300 nt)和 hpCMV2 + 1 (hpCMV2和hpCMV1融合)片段），随后通过农杆菌介导的瞬时转化法加以确认。其中，通过hpCMV2 + 1载体表达的dsRNAs对CMV-Ca-P1和CMV-Fny（普通菌株）均表现出抗性。为了获得对CMV-Ca-P1和CMV-Fny均有较高的抗性，本研究开发含有hpCMV2 + 1载体的转基因烟草，并使其具有抵抗CMV-Ca-P1和CMV-Fny的抗性。本研究有助于可抑制CMV侵染靶序列的有效筛选。
外文摘要：Cucumber mosaic virus Ca-P1 strain, a P0 resistance-breaking virus, was isolated from the leaves of virus-infected Capsicum annuum 'Manidda' (a P0 resistant cultivar) in South Korea. Since CMV-Ca-P1 was first reported in 2006, this virus causing damage on pepper production has been constantly detected in South Korea. We constructed three CMV RNAi vectors based on the post-transcriptional gene silencing of defense system against virus infection. These independent vectors (hpCMV1, hpCMV2, and hpCMV2 + 1 RNAi vectors) were designed to produce dsRNAs containing each hpCMV1 (1270-1629 nt), hpCMV2 (1-300 nt), and hpCMV2 + 1 (fused hpCMV2 and hpCMV1) fragments, in RNA-1 (replicase gene) of CMV-Ca-P1, which were then confirmed by Agrobacterium-mediated transformation transient assay. Among these, dsRNAs expressed from the hpCMV2 + 1 vector showed resistance to both CMV-Ca-P1 and CMV-Fny (ordinary strain). To obtain high level of resistance to both CMV-Ca-P1 and CMV-Fny, transgenic Nicotiana benthamiana plants containing hpCMV2 + 1 vector were developed and conferred resistance to both CMV-Ca-P1 and CMV-Fny. This study contributes to the effective selection of target sequences that may inhibit CMV infection.
外文关键词：Cucumber mosaic virus; Transgenic plant; Agrobacterium-mediated transformation transient assay CMV RNAi vector; Post-transcriptional gene silencing; siRNA
作者：Song, Eun Gyeong; Ryu, Ki Hyun
作者单位：首尔女子大学
期刊名称：PLANT BIOTECHNOLOGY REPORTS
期刊影响因子：1.422
出版年份：2017
出版刊次：12
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