Biomedical Research Center, Institute of Virology , Slovak Academy of Sciences , Dubravska cesta 9 , 845 05 Bratislava , Slovak Republic.
Institute of Microbiology , The Czech Academy of Sciences , Videnska 1083 , 142 20 Prague , Czech Republic.
J Proteome Res. 2018 Sep 7;17(9):3114-3127. doi: 10.1021/acs.jproteome.8b00230. Epub 2018 Aug 22.
Plum pox virus (PPV, family Potyviridae) is one of the most important viral pathogens of Prunus spp. causing considerable damage to stone-fruit industry worldwide. Among the PPV strains identified so far, only PPV-C, PPV-CR, and PPV-CV are able to infect cherries under natural conditions. Herein, we evaluated the pathogenic potential of two viral isolates in herbaceous host Nicotiana benthamiana. Significantly higher accumulation of PPV capsid protein in tobacco leaves infected with PPV-CR (RU-30sc isolate) was detected in contrast to PPV-C (BY-101 isolate). This result correlated well with the symptoms observed in the infected plants. To further explore the host response upon viral infection at the molecular level, a comprehensive proteomic profiling was performed. Using reverse-phase ultra-high-performance liquid chromatography followed by label-free mass spectrometry quantification, we identified 38 unique plant proteins as significantly altered due to the infection. Notably, the abundances of photosynthesis-related proteins, mainly from the Calvin-Benson cycle, were found more aggressively affected in plants infected with PPV-CR isolate than those of PPV-C. This observation was accompanied by a significant reduction in the amount of photosynthetic pigments extracted from the leaves of PPV-CR infected plants. Shifts in the abundance of proteins that are involved in stimulation of photosynthetic capacity, modification of amino acid, and carbohydrate metabolism may affect plant growth and initiate energy formation via gluconeogenesis in PPV infected N. benthamiana. Furthermore, we suggest that the higher accumulation of HO in PPV-CR infected leaves plays a crucial role in plant defense and development by activating the glutathione synthesis.
李痘病毒(PPV,壶菌病毒科)是李属植物的最重要的病毒病原体之一,给全球核果产业造成了相当大的损失。到目前为止,在已鉴定的 PPV 株系中,只有 PPV-C、PPV-CR 和 PPV-CV 能够在自然条件下感染樱桃。在此,我们评估了两种病毒分离物在草本宿主烟草原生质体中的致病潜能。与 PPV-C(BY-101 分离物)相比,感染 PPV-CR(RU-30sc 分离物)的烟草叶片中 PPV 衣壳蛋白的积累明显更高。这一结果与感染植物中观察到的症状很好地相关。为了进一步在分子水平上探索病毒感染时宿主的反应,我们进行了全面的蛋白质组学分析。使用反相超高效液相色谱法结合无标记质谱定量法,我们鉴定出 38 种独特的植物蛋白因感染而显著改变。值得注意的是,感染 PPV-CR 分离物的植物中,与光合作用相关的蛋白质(主要来自卡尔文-本森循环)的丰度受到更强烈的影响,而感染 PPV-C 的植物则不然。这一观察结果伴随着从感染 PPV-CR 的植物叶片中提取的光合色素量的显著减少。参与刺激光合作用能力、氨基酸修饰和碳水化合物代谢的蛋白质丰度的变化可能会影响 PPV 感染的烟草原生质体的生长,并通过糖异生启动能量形成。此外,我们认为 PPV-CR 感染叶片中 HO 的更高积累通过激活谷胱甘肽合成在植物防御和发育中发挥关键作用。