Massively parallel sequencing of the diapause transcriptome in Aedes albopictus

描述（申请人提供）：亚洲虎蚊白纹伊蚊（Aedes albopictus, Skuse）是登革热的重要媒介，也是黄热病、基孔肯雅病毒和几种北美原生虫媒病毒的有效传播媒介。在过去的20年里，这种蚊子从其原产于亚洲的范围迅速传播到全球，最近它参与了疾病暴发，这是一个突出的公共卫生问题。光周期性介导的卵滞育是白纹伊蚊和其他医学相关蚊子在温带环境中生存和传播的关键生态特征。本研究计划的长期目标是严格定义白纹伊蚊光周期滞育的分子基础，作为基于破坏这一关键生态适应开发新型媒介控制形式的基础。本提案的直接目标是确定白纹伊蚊光周期滞育反应的转录因子。在Specific Aim 1中，大规模平行焦磷酸测序和生物信息学分析将用于比较卵母细胞、发育中的胚胎和pharate幼虫的滞育与非滞育转录组。在Specific Aim 2中，定量RT- PCR将通过比较滞育（温带）和非滞育（热带）种群之间的基因表达来确定支撑滞育反应的转录事件。这些研究结果除了为开发新型病媒控制手段提供基础外，还将迅速推进蚊子基因发现和应激反应生理学的理解。光周期滞育是指昆虫测量白天长度（光周期）的能力，作为启动季节性发育停滞模式的线索。本研究将探讨白纹伊蚊（Aedes albopictus）光周期滞育的分子调控。这项研究的结果将为基于这种关键生态适应的破坏制定新的病媒控制策略提供基础。

DESCRIPTION (provided by applicant): The Asian tiger mosquito, Aedes albopictus (Skuse), is an important vector of dengue fever and also efficiently transmits yellow fever, chikungunya virus and several native North American arboviruses. The rapid spread of this mosquito from its native Asian range across the globe during the last 20 years and its recent involvement in disease outbreaks represents an outstanding public health concern. Photoperiodically mediated egg diapause is a pivotal ecological trait central to the ability of A. albopictus and other medically relevant mosquitoes to inhabit temperate environments and spread across broad geographic ranges. The long-term goal of this research program is to rigorously define the molecular basis of photoperiodic diapause in A. albopictus as a foundation for developing novel forms of vector control based on the disruption of this critical ecological adaptation. The immediate goal of this proposal is to identify transcriptional elements of the photoperiodic diapause response in A. albopictus. In Specific Aim One, massively parallel pyrosequencing and bioinformatics analyses will be used to compare diapause vs. non-diapause transcriptomes from oocytes, developing embryos, and pharate larvae. In Specific Aim 2, quantitative RT- PCR will be employed to identify transcriptional events underpinning the diapause response by comparing gene expression between a diapausing (temperate) and non- diapausing (tropical) population. In addition to providing a foundation for developing novel forms of vector control, results from these studies will rapidly advance gene discovery and understanding of stress response physiology in mosquitoes. PUBLIC HEALTH RELEVANCE Photoperiodic diapause refers to the ability of insects to measure day length (photoperiod) as a cue for initiating seasonally appropriate patterns of developmental arrest. This study will investigate the molecular control of photoperiodic diapause in the invasive and medically important mosquito, Aedes albopictus. Results of this study will provide a foundation for developing novel strategies for vector control based on the disruption of this critical ecological adaptation.