已通过宏基因组学/超转录组学方法发现了编码单链DNA病毒的环状复制相关蛋白的未表征的病毒基因组。这些新病毒中的一些被分类为新形成的基因组病毒科。这里,我们确定了一种新型基因病毒的宿主范围,SlaGemV-1,通过感染克隆转染菌核病菌。接种获救的病毒体,我们进一步转染了灰葡萄孢菌和果霉病,Screrotiniaceae家族的两个经济上重要的成员,和尖孢镰刀菌.SlaGemV-1在硬化链球菌中引起低毒力,B.cinerea,和M.Fructicola.SlaGemV-1也在斜纹夜蛾昆虫细胞中复制,但在秀丽隐杆线虫或植物中不复制。通过位点特异性整合分别表达病毒基因,仅复制蛋白就足以引起衰弱。我们的研究是第一个证明在没有已知宿主的情况下重建宏基因发现的基因组病毒,具有诱导低毒力的潜力,感染性克隆允许研究跨属保守的基因组病毒与宿主相互作用的机制。重要性关于广泛的基因病毒的确切宿主范围知之甚少。大豆叶相关的gemygorvirus-1(SlaGemV-1)的基因组最初是在没有已知宿主的情况下从宏基因组/转移基因组研究中组装而成的。这里,我们拯救了SlaGemV-1,发现它可以感染三种重要的植物病原真菌和秋季粘虫(S.frugiperdaSf9)昆虫细胞,但不是模型线虫,C.秀丽隐杆线虫,或模型植物物种。最重要的是,SlaGemV-1显示出有望诱导菌科中测试真菌物种的低毒力,包括菌核病,灰葡萄孢菌,和果糖莫尼氏菌。低毒力的病毒决定簇被进一步鉴定为复制起始蛋白。作为概念的证明,我们证明,当新病毒被拯救并表征其宿主范围时,在植物宏基因组中发现的病毒可以是有价值的遗传资源。
Uncharacterized viral genomes that encode circular replication-associated proteins of single-stranded DNA viruses have been discovered by metagenomics/metatranscriptomics approaches. Some of these novel viruses are classified in the newly formed family Genomoviridae. Here, we determined the host range of a novel
genomovirus, SlaGemV-1, through the transfection of Sclerotinia sclerotiorum with infectious clones. Inoculating with the rescued virions, we further transfected Botrytis cinerea and Monilinia fructicola, two economically important members of the family Sclerotiniaceae, and Fusarium oxysporum. SlaGemV-1 causes hypovirulence in S. sclerotiorum, B. cinerea, and M. fructicola. SlaGemV-1 also replicates in Spodoptera frugiperda insect cells but not in Caenorhabditis elegans or plants. By expressing viral genes separately through site-specific integration, the replication protein alone was sufficient to cause debilitation. Our study is the first to demonstrate the reconstruction of a metagenomically discovered
genomovirus without known hosts with the potential of inducing hypovirulence, and the infectious clone allows for studying mechanisms of
genomovirus-host interactions that are conserved across genera. IMPORTANCE Little is known about the exact host range of widespread genomoviruses. The genome of soybean leaf-associated gemygorvirus-1 (SlaGemV-1) was originally assembled from a metagenomic/metatranscriptomic study without known hosts. Here, we rescued SlaGemV-1 and found that it could infect three important plant-pathogenic fungi and fall armyworm (S. frugiperda Sf9) insect cells but not a model nematode, C. elegans, or model plant species. Most importantly, SlaGemV-1 shows promise for inducing hypovirulence of the tested fungal species in the family Sclerotiniaceae, including Sclerotinia sclerotiorum, Botrytis cinerea, and Monilinia fructicola. The viral determinant of hypovirulence was further identified as replication initiation protein. As a proof of concept, we demonstrate that viromes discovered in plant metagenomes can be a valuable genetic resource when novel viruses are rescued and characterized for their host range.