Temperature dependent impacts of RNA virus population evolution

摘要 节肢动物传播病毒（虫媒病毒）的出现是一个持续的公共卫生问题。条件 导致新病毒基因型出现的原因还知之甚少。在蚊子中， 通过复制率和病毒从病毒传播的可能性， 中肠并最终传播。然而，在关于外在孵化的影响的文献中存在空白， (EI)温度对蚊子体内病毒的多样性有影响。因此，我们计划研究 温度在虫媒病毒的进化中起作用。首先，我们将确定EI在多大程度上 温度影响伊蚊和库蚊中寨卡病毒（ZIKV）和西尼罗河病毒（WNV）的载体能力 蚊子，并在此之后使用RNA-seq来确定共有基因组如何改变， 对EI温度的响应。接下来，我们将评估温度驱动的单核苷酸变体， 进行群体遗传学分析，以确定EI温度对病毒群体的影响程度 多样性这可以提供病毒进化的温度特异性模型。最后，我们将验证 温度驱动的病毒进化模型，将其与当地传播的WNV田间分离株进行比较。的 这些研究的结果将为虫媒病毒的进化提供见解，并可能用于预测 环境条件可能导致新病毒基因型的进化和选择， 爆发。 假设：我们假设，当虫媒病毒如WNV和ZIKV经历外源性孵育时， 它们各自的载体，高温孵育驱动病毒群体多样性，导致突变 haplotypes正在传播。这一假设将在三个具体目标进行检验 目标1。确定外源孵育温度影响ZIKV和WNV载体的程度 在蚊子中的竞争力，并确定任何温度驱动的一致和可预测的共识基因组 变化 目标2.确定外源孵育温度如何影响ZIKV和WNV群体多样性 在蚊子体内。 目标3：描述2013-2017年柯林斯堡温度对西尼罗河病毒演变的影响。

Abstract The emergence of arthropod-borne viruses (arboviruses) is an ongoing public health concern. The conditions that lead to the emergence of new virus genotypes are poorly understood. In mosquitoes, temperature profoundly impacts virus transmission via replication rate and the likelihood that virus will disseminate from the midgut and ultimately be transmitted. Yet, there is a gap in the literature on the effect that extrinsic incubation (EI) temperature has on virus diversification in the mosquito. Therefore, we plan to study the role that temperature plays in arbovirus evolution during the EI period. First, we will determine the extent to which EI temperature impacts Zika virus (ZIKV) and West Nile virus (WNV) vector competence in Aedes and Culex mosquitoes respectively, and following this use RNA-seq to identify how the consensus genome changes in response to the temperature of EI. Next, we will assess temperature driven single-nucleotide variants and conduct population genetics analysis to determine the extent of the EI temperature’s effect on viral population diversity. This may provide a temperature specific model of viral evolution. Last, we will validate our temperature driven, viral evolution model by comparing it to locally transmitted WNV field isolates. The outcome of these studies will provide insight to arbovirus evolution and potentially be used to predict environmental conditions that may lead to evolution and selection of novel virus genotypes that cause severe outbreaks. Hypothesis: We hypothesis that when arboviruses such as WNV and ZIKV undergo extrinsic incubation in their respective vector, a high temperature of incubation drives viral population diversity, leading to mutant haplotypes being transmitted. This hypothesis will be tested in three specific aims Aim 1. Determine the extent in which the temperature of extrinsic incubation impacts ZIKV and WNV vector competence in mosquitoes and identify any temperature driven consistent and predictable consensus genome changes. Aim 2. Determine how the temperature of extrinsic incubation influences ZIKV and WNV population diversity within mosquitoes. Aim 3. Characterize the effects of temperature on WNV evolution in Fort Collins from 2013-2017.