Interspecies/Intergroup Complementation of Orthotospovirus Replication and Movement through Reverse Genetics Systems
Interspecies/Intergroup Complementation of Orthotospovirus Replication and Movement through Reverse Genetics Systems
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DOI：
10.1128/jvi.01809-22
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发表时间：
2023-04-06
影响因子：
5.4
通讯作者：
Tao，Xiaorong
中科院分区：
文献类型：
--
作者：
Feng，Mingfeng;Chen，Minglong;Tao，Xiaorong
正形体病毒，即感染植物的布尼亚病毒，在农作物中造成严重疾病，并对全球粮食安全构成重大威胁。正畸病毒家族包含30多个成员，可分为两个地理组，美洲型和欧洲/亚洲型正畸病毒。然而，不同物种之间的遗传相互作用以及在混合感染期间，来自不同地理群体的正孢子病毒基因功能的易位互补的可能性仍未得到充分探讨。本研究建立了美国凤仙花坏死斑病毒（INSV）和马蹄莲绿斑病毒（CCSV）、番茄带状斑点病毒（TZSV）（两种具有代表性的欧洲/亚洲正形孢子病毒）的微型复制型反向遗传（RG）系统。结合早前建立的番茄斑点枯萎病毒（TSWV） RG系统，进行了病毒复制酶/运动蛋白的交换，并对其进行了种间转互补分析。同源RNA依赖RNA聚合酶（RdRp）和核衣壳(N)蛋白支持两个地理组的正孢子病毒复制，而来自一组的RdRp和来自另一组的N的异源组合不能支持两个地理组的病毒复制。此外，NSm运动蛋白（MP），来自两个地理群的正形孢子病毒，能够在其运动中对异源正形孢子病毒或正链黄瓜花叶病毒（CMV）进行转补体，尽管效率不同。来自稻条细纹病毒（RSV）的MP，一种不同于正形孢子病毒的感染植物的布雅病毒，或来自巨细胞病毒的MP也移动正形孢子病毒。我们的研究结果对植物正畸病毒的遗传相互作用/重组潜力有了深入的了解。正形孢子病毒是农业上重要的负链RNA病毒，在世界范围内对许多作物造成严重的产量损失。尽管新的感染动物的布尼亚病毒的出现通常与基因重组有关，但这一问题在感染植物的正形体病毒中仍然没有得到充分的关注。随着不同地理区域正形孢子病毒反向遗传系统的建立，研究了美洲型和欧洲/亚洲型正形孢子病毒的种间/群间复制/运动互补。美洲正形孢子病毒基因组rna可被欧洲/亚洲正形孢子病毒基因组rna的RdRp和N复制，反之亦然。然而，它们的基因组rna不能通过来自一个地理群体的RdRp和来自另一个地理群体的N的异源组合来复制。病毒实体在细胞间的运动得到来自两个地理群的NSm的支持，其中来自同一群病毒的NSm效率最高。我们的发现为了解不同种类的正形体病毒之间的基因相互作用和病毒基因功能的交换能力提供了重要的见解。
Orthotospoviruses, the plant-infecting bunyaviruses, cause serious diseases in agronomic crops and pose major threats to global food security. The family ofTospoviridaecontains more than 30 members that are classified into two geographic groups, American-type and Euro/Asian-type orthotospovirus. However, the genetic interaction between different species and the possibility, during mixed infections, for transcomplementation of gene functions by orthotospoviruses from different geographic groups remains underexplored. In this study, minireplicon-based reverse genetics (RG) systems have been established forImpatiens necrotic spot virus(INSV) (an American-type orthotospovirus) and forCalla lily chlorotic spot virusandTomato zonate spot virus(CCSV and TZSV) (two representative Euro/Asian orthotospoviruses). Together with the earlier established RG system forTomato spotted wilt virus(TSWV), a type species of theOrthotospovirusAmerican-clade, viral replicase/movement proteins were exchanged and analyzed on interspecies transcomplementation. Whereas the homologous RNA-dependent RNA polymerase (RdRp) and nucleocapsid (N) protein supported the replication of orthotospoviruses from both geographic groups, heterologous combinations of RdRp from one group and N from the other group were unable to support the replication of viruses from both groups. Furthermore, the NSm movement protein (MP), from both geographic groups of orthotospoviruses, was able to transcomplement heterologous orthotospoviruses or a positive-strandCucumber mosaic virus(CMV) in their movement, albeit with varying efficiency. MP fromRice stripe tenuivirus(RSV), a plant-infecting bunyavirus that is distinct from orthotospoviruses, or MP from CMV also moves orthotospoviruses. Our findings gain insights into the genetic interaction/reassortant potentials for the segmented plant orthotospoviruses.IMPORTANCEOrthotospoviruses are agriculturally important negative-strand RNA viruses and cause severe yield-losses on many crops worldwide. Whereas the emergence of new animal-infecting bunyaviruses is frequently associated with genetic reassortants, this issue remains underexposed with the plant-infecting orthotospovirus. With the development of reverse genetics systems for orthotospoviruses from different geographic regions, the interspecies/intergroup replication/movement complementation between American- and Euro/Asian-type orthotospoviruses were investigated. Genomic RNAs from American orthotospoviruses can be replicated by the RdRp and N from those of Euro/Asia-group orthotospoviruses, and vice versa. However, their genomic RNAs cannot be replicated by a heterologous combination of RdRp from one geographic group and N from another geographic group. Cell-to-cell movement of viral entity is supported by NSm from both geographic groups, with highest efficiency by NSm from viruses belonging to the same group. Our findings provide important insights into the genetic interaction and exchange ability of viral gene functions between different species of orthotospovirus.