Cucumber RDR1s and cucumber mosaic virus suppressor protein 2b association directs host defence in cucumber plants,Molecular Plant Pathology

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Cucumber RDR1s and cucumber mosaic virus suppressor protein 2b association directs host defence in cucumber plants
Molecular Plant Pathology ( IF 4.8 ) Pub Date : 2021-08-06 , DOI: 10.1111/mpp.13112
Reenu Kumari _{1,

2} , Surender Kumar _{1,

3} , Diana Leibman ₁ , Bekele Abebie ₁ , Yulia Shnaider ₁ , Shou-Wei Ding ₄ , Amit Gal-On ₁

Affiliation

RNA-dependent RNA polymerases (RDRs) regulate important aspects of plant development and resistance to pathogens. The role of RDRs in virus resistance has been demonstrated using siRNA signal amplification and through the methylation of viral genomes. Cucumber (Cucumis sativus) has four RDR1 genes that are differentially induced during virus infection: CsRDR1a, CsRDR1b, and duplicated CsRDR1c1/c2. The mode of action of CsRDR1s during viral infection is unknown. Transient expression of the cucumber mosaic virus (CMV)-2b protein (the viral suppressor of RNA silencing) in cucumber protoplasts induced the expression of CsRDR1c, but not of CsRDR1a/1b. Results from the yeast two-hybrid system showed that CsRDR1 proteins interacted with CMV-2b and this was confirmed by bimolecular fluorescence complementation assays. In protoplasts, CsRDR1s localized in the cytoplasm as punctate spots. Colocalization experiments revealed that CsRDR1s and CMV-2b were uniformly dispersed throughout the cytoplasm, suggesting that CsRDR1s are redistributed as a result of interactions. Transient overexpression of individual CsRDR1a/1b genes in protoplasts reduced CMV accumulation, indicating their antiviral role. However, overexpression of CsRDR1c in protoplasts resulted in relatively higher accumulation of CMV and CMVΔ2b. In single cells, CsRDR1c enhances viral replication, leading to CMV accumulation and blocking secondary siRNA amplification of CsRDR1c by CMV-2b protein. This suggests that CMV-2b acts as both a transcription factor that induces CsRDR1c (controlling virus accumulation) and a suppressor of CsRDR1c activity.

中文翻译：

黄瓜 RDR1s 和黄瓜花叶病毒抑制蛋白 2b 联合指导黄瓜植物的宿主防御

RNA 依赖性 RNA 聚合酶 (RDR) 调节植物发育和病原体抗性的重要方面。RDRs 在病毒抗性中的作用已通过 siRNA 信号放大和病毒基因组的甲基化得到证实。黄瓜 ( Cucumis sativus ) 有四个RDR1基因，它们在病毒感染期间被差异诱导：CsRDR1a、CsRDR1b和重复的 CsRDR1c1 / c2。CsRDR1s 在病毒感染期间的作用方式尚不清楚。黄瓜原生质体中黄瓜花叶病毒 (CMV)-2b 蛋白（RNA 沉默的病毒抑制因子）的瞬时表达诱导CsRDR1c的表达，但不诱导 CsRDR1c 的表达。CsRDR1a/1b。酵母双杂交系统的结果表明，CsRDR1 蛋白与 CMV-2b 相互作用，这已通过双分子荧光互补测定得到证实。在原生质体中，CsRDR1s 定位在细胞质中作为点状斑点。共定位实验表明，CsRDR1s 和 CMV-2b 均匀分散在整个细胞质中，表明 CsRDR1s 由于相互作用而重新分布。个体CsRDR1a/1b的瞬时过表达原生质体中的基因减少了 CMV 的积累，表明它们具有抗病毒作用。然而，原生质体中 CsRDR1c 的过表达导致 CMV 和 CMVΔ2b 相对较高的积累。在单个细胞中，CsRDR1c 增强病毒复制，导致 CMV 积累并阻断 CsRDR1c 由 CMV-2b 蛋白对 CsRDR1c 的二级 siRNA 扩增。这表明 CMV-2b 既可作为诱导 CsRDR1c（控制病毒积累）的转录因子，又可作为 CsRDR1c 活性的抑制剂。

更新日期：2021-10-15

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