Characterization of a Brome mosaic virus strain and its use as a vector for gene silencing in monocotyledonous hosts
Characterization of a Brome mosaic virus strain and its use as a vector for gene silencing in monocotyledonous hosts
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DOI：
10.1094/mpmi-19-1229
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发表时间：
2006-11-01
影响因子：
3.5
通讯作者：
Nelson, Richard S.
中科院分区：
文献类型：
--
作者：
Ding, Xin Shun;Schneider, William L.;Nelson, Richard S.
病毒诱导的基因沉默（VIGS）用于分析双子叶植物的基因功能，但在单子叶植物（特别是水稻和玉米）中的应用较少，部分原因是可用的病毒表达载体数量有限。在这里，我们报告了来自羊茅的病毒 VIGS 的克隆和修饰。 （高羊茅）在温室条件下引起大麦、水稻和特定玉米品种（Va35）的系统性花叶症状。经测序，确定该病毒为雀麦花叶病毒（BMV）毒株。该病毒被命名为 F-BMV（F 代表羊茅），控制水稻全身感染的遗传决定子被映射到三方基因组的 RNA I 和 2 上。来自 BMV 俄罗斯毒株 (R-BMV) 的 RNA 3 的 cDNA 被修改以接受来自外源基因的插入。将表达部分植物基因的 F-BMV 的 RNA I 和 2 以及 R-BMV 的 RNA 3 共接种到大麦、水稻和玉米植物的叶子上，导致视觉沉默样表型。视觉表型与相应叶子中靶宿主转录物水平降低相关。 VIGS 视觉表型从肌动蛋白 1 转录物表达沉默期间维持的变化到八氢番茄红素去饱和酶表达沉默期间不完全渗透受影响组织的短暂变化。 F-BMV RNA 3 经过修饰，可以增加病毒的积累，同时最大限度地减少病毒的致病性。修饰后的载体 C-BMVA/G（C 表示嵌合）被证明可用于 VIGS。这些 BMV 载体将可用于分析水稻和玉米中的基因功能，但尚未有 VIGS 系统的报道。
Virus-induced gene silencing (VIGS) is used to analyze gene function in dicotyledonous plants but less so in monocotyledonous plants (particularly rice and corn), partially due to the limited number of virus expression vectors available. Here, we report the cloning and modification for VIGS of a virus from Festuca arundinacea Schreb. (tall fescue) that caused systemic mosaic symptoms on barley, rice, and a specific cultivar of maize (Va35) under greenhouse conditions. Through sequencing, the virus was determined to be a strain of Brome mosaic virus (BMV). The virus was named F-BMV (F for Festuca), and genetic determinants that controlled the systemic infection of rice were mapped to RNAs I and 2 of the tripartite genome. cDNA from RNA 3 of the Russian strain of BMV (R-BMV) was modified to accept inserts from foreign genes. Coinoculation of RNAs I and 2 from F-BMV and RNA 3 from R-BMV expressing a portion of a plant gene to leaves of barley, rice, and maize plants resulted in visual silencing-like phenotypes. The visual phenotypes were correlated with decreased target host transcript levels in the corresponding leaves. The VIGS visual phenotype varied from maintained during silencing of actin 1 transcript expression to transient with incomplete penetration through affected tissue during silencing of phytoene desaturase expression. F-BMV RNA 3 was modified to allow greater accumulation of virus while minimizing virus pathogenicity. The modified vector C-BMVA/G (C for chimeric) was shown to be useful for VIGS. These BMV vectors will be useful for analysis of gene function in rice and maize for which no VIGS system is reported.