Panno Stefano, Caruso Andrea Giovanni, Bertacca Sofia, Matić Slavica, Davino Salvatore, Parrella Giuseppe
Department of Agricultural, Food and Forest Sciences, University of Palermo, Palermo, Italy.
Department of Biological, Chemical and Pharmaceutical Science and Technologies, University of Palermo, Palermo, Italy.
Front Plant Sci. 2021 Sep 3;12:698573. doi: 10.3389/fpls.2021.698573. eCollection 2021.
Parietaria mottle virus (PMoV) is considered an emerging virus in many countries of the Mediterranean basin, especially on tomato and pepper crops. Symptoms on tomato leaves and fruits can be easily confused with those induced by cucumber mosaic virus (CMV) with necrogenic satellite RNA (CMV-satRNA), tomato spotted wilt virus (TSWV) or tomato mosaic virus (ToMV). Mixed infection of these viruses has been also reported in some tomato cultivars, with an increase in the complexity of the symptoms and severity of the disease. Although a specific serum and riboprobes have been produced, nowadays no sensitive diagnostic methods are available for the rapid PMoV detection. Here, we have developed a RT-qPCR assay with the aim to establish a more sensitive and specific method for PMoV detection. Specific primers and TaqMan probe were designed and tested with all PMoV isolates available in GenBank. Moreover, this method was evaluated on tomato naturally infected samples from Sicily region (Italy). Results obtained showed that the RT-qPCR assay developed in this work is extremely sensitive, in fact, it is able to detect as few as 10 PMoV RNA copies in tomato total RNA; moreover, it will be a particularly valuable tool for early detection of PMoV. Furthermore, the analyzes on field samples show how this pathogen is increasingly present in tomato crops in the last years, helping to undermine the Italian horticultural sector.
墙草斑驳病毒(PMoV)在地中海盆地的许多国家被视为一种新兴病毒,尤其是在番茄和辣椒作物上。番茄叶片和果实上的症状很容易与由黄瓜花叶病毒(CMV)及其坏死性卫星RNA(CMV-satRNA)、番茄斑萎病毒(TSWV)或番茄花叶病毒(ToMV)引起的症状相混淆。在一些番茄品种中也报道了这些病毒的混合感染,这增加了症状的复杂性和疾病的严重程度。尽管已经制备了特异性血清和核糖探针,但目前尚无用于快速检测PMoV的灵敏诊断方法。在此,我们开发了一种RT-qPCR检测方法,旨在建立一种更灵敏、特异的PMoV检测方法。设计了特异性引物和TaqMan探针,并对GenBank中所有可用的PMoV分离株进行了测试。此外,该方法还在来自意大利西西里岛地区的番茄自然感染样本上进行了评估。所获得的结果表明,本研究中开发的RT-qPCR检测方法极其灵敏,事实上,它能够在番茄总RNA中检测到低至10个PMoV RNA拷贝;此外,它将是早期检测PMoV的一个特别有价值的工具。此外,对田间样本的分析表明,近年来这种病原体在番茄作物中越来越普遍,这对意大利园艺业造成了损害。