Using a Viral Vector to Reveal the Role of MicroRNA159 in Disease Symptom Induction by a Severe Strain of Cucumber mosaic virus
Using a Viral Vector to Reveal the Role of MicroRNA159 in Disease Symptom Induction by a Severe Strain of Cucumber mosaic virus
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使用病毒载体揭示 MicroRNA159 在黄瓜花叶病毒重毒株诱导疾病症状中的作用
DOI：
10.1104/pp.113.232090
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发表时间：
2014-03-01
期刊：
影响因子：
7.4
通讯作者：
Carr, John P.
中科院分区：
文献类型：
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作者：
Du, Zhiyou;Chen, Aizhong;Carr, John P.
在转基因拟南芥（Arabidopsis thaliana）中，来自严重IA亚群菌株Fny的黄瓜花叶病毒（CMV） 2b沉默抑制蛋白的表达破坏了microRNA （miRNA）的发育，但来自轻度II亚群菌株（Q和LS）的同源物却没有，这解释了症状严重程度的菌株特异性差异。然而，目前尚不清楚哪些受Fny2b影响的mirna会严重影响病毒症状。Fny2b转基因植物表型microRNA159ab （mir159ab）突变植物以及Fny2b改变mir159ab调控的转录物水平的观察表明，mir159ab在Fny-CMV引发严重症状中的作用。利用在表达人工miRNA的转基因植物中恢复正常表型作为概念的证明，我们开发了一种基于ls - cmv的载体来表达模仿miRNA目标的序列。使用LS-CMV耗尽的miR159表达miR159靶模拟序列，诱导类似Fny-CMV的症状。在含有miR159ab靶点MYB DOMAIN PROTEIN33 （MYB33）和MYB65突变等位基因的植物中抑制fny - cmv诱导的症状证实了该miRNA在发病机制中的重要性。本研究证明了利用病毒载体表达miRNA靶模拟物，促进植物miRNA功能研究的实用性。
In transgenic Arabidopsis (Arabidopsis thaliana), expression of the Cucumber mosaic virus (CMV) 2b silencing suppressor protein from the severe subgroup IA strain Fny disrupted microRNA (miRNA)-regulated development but orthologs from mild subgroup II strains (Q and LS) did not, explaining strain-specific differences in symptom severity. However, it is unknown which miRNAs affected by Fny2b critically affect viral symptoms. Observations that Fny2b-transgenic plants phenocopy microRNA159ab (mir159ab) mutant plants and that Fny2b altered miR159ab-regulated transcript levels suggested a role for miR159ab in elicitation of severe symptoms by Fny-CMV. Using restoration of the normal phenotype in transgenic plants expressing an artificial miRNA as a proof of concept, we developed a LS-CMV-based vector to express sequences mimicking miRNA targets. Expressing a miR159 target mimic sequence using LS-CMV depleted miR159 and induced symptoms resembling those of Fny-CMV. Suppression of Fny-CMV-induced symptoms in plants harboring mutant alleles for the miR159ab targets MYB DOMAIN PROTEIN33 (MYB33) and MYB65 confirmed the importance of this miRNA in pathogenesis. This study demonstrates the utility of a viral vector to express miRNA target mimics to facilitate functional studies of miRNAs in plants.