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Abstract：
Viruses are factors that can fluctuate insect populations, including honey bees. Most honey bee infecting viruses are single positive-stranded RNA viruses that may not specifically infect honey bees and can be hazardous to other pollinator insects. In addition, these viruses could synergize with other stressors to worsen the honey bee population decline. To identify the underlying detailed mechanisms, reversed genetic studies with infectious cDNA clones of the viruses are necessary. Moreover, an infectious cDNA clone can be applied to studies as an ideal virus isolate that consists of a single virus species with a uniform genotype. However, only a few infectious cDNA clones have been reported in honey bee studies since the first infectious cDNA clone was published four decades ago. This article discusses steps, rationales, and potential issues in bee-infecting RNA virus cloning. In addition, failed experiences of cloning a Deformed wing virus isolate that was phylogenetically identical to Kakugo virus were addressed. We hope the information provided in this article can facilitate further developments of reverse-genetic studies of bee-infecting viruses to clarify the roles of virus diseases in the current pollinator declines.
摘要：
病毒是昆虫种群波动的因素,包括蜜蜂。大多数蜜蜂感染病毒是单链RNA病毒，可能不会特异性感染蜜蜂，并且可能对其他传粉昆虫有害。此外,这些病毒可能与其他压力因素协同作用，加剧蜜蜂种群的减少。为了确定潜在的详细机制，用病毒的感染性cDNA克隆进行反向遗传研究是必要的。此外，感染性cDNA克隆可以作为理想的病毒分离株用于研究，该分离株由具有统一基因型的单一病毒种组成。然而,自从40年前第一个感染性cDNA克隆发表以来，在蜜蜂研究中只报道了少数感染性cDNA克隆。本文讨论了步骤，理据,以及蜜蜂感染RNA病毒克隆中的潜在问题。此外,解决了克隆与Kakugo病毒在系统发育上相同的变形机翼病毒分离株的失败经验。我们希望本文提供的信息可以促进蜜蜂感染病毒的反向遗传研究的进一步发展，以阐明病毒疾病在当前传粉媒介下降中的作用。
