Department of Entomology, Max Planck Institute for Chemical Ecology, Jena, Germany.
Insect Mol Biol. 2014 Feb;23(1):98-112. doi: 10.1111/imb.12067. Epub 2013 Nov 20.
The pollen beetle (Meligethes aeneus) is a major pest of oilseed rape (Brassica napus) and other cruciferous crops in Europe. Pesticide-resistant pollen beetle populations are emerging, increasing the economic impact of this species. We isolated total RNA from the larval and adult stages, the latter either naïve or immunized by injection with bacteria and yeast. High-throughput RNA sequencing (RNA-Seq) was carried out to establish a comprehensive transcriptome catalogue and to screen for developmental stage-specific and immunity-related transcripts. We assembled the transcriptome de novo by combining sequence tags from all developmental stages and treatments. Gene expression data based on normalized read counts revealed several functional gene categories that were differentially expressed between larvae and adults, particularly genes associated with digestion and detoxification that were induced in larvae, and genes associated with reproduction and environmental signalling that were induced in adults. We also identified many genes associated with microbe recognition, immunity-related signalling and defence effectors, such as antimicrobial peptides (AMPs) and lysozymes. Digital gene expression analysis revealed significant differences in the profile of AMPs expressed in larvae, naïve adults and immune-challenged adults, providing insight into the steady-state differences between developmental stages and the complex transcriptional remodelling that occurs following the induction of immunity. Our data provide insight into the adaptive mechanisms used by phytophagous insects and could lead to the development of more effective control strategies for insect pests.
花粉象甲（Meligethes aeneus）是欧洲油菜（Brassica napus）和其他十字花科作物的主要害虫。抗药性花粉象甲种群正在出现，这增加了该物种的经济影响。我们从幼虫和成虫阶段分离了总 RNA，后者要么是未免疫的，要么是通过注射细菌和酵母免疫的。进行了高通量 RNA 测序（RNA-Seq），以建立一个全面的转录组目录，并筛选发育阶段特异性和免疫相关的转录本。我们通过组合来自所有发育阶段和处理的序列标签从头组装转录组。基于归一化读取计数的基因表达数据揭示了幼虫和成虫之间差异表达的几个功能基因类别，特别是在幼虫中诱导的与消化和解毒相关的基因，以及在成虫中诱导的与繁殖和环境信号相关的基因。我们还鉴定了许多与微生物识别、免疫相关信号和防御效应子相关的基因，如抗菌肽 (AMP) 和溶菌酶。数字基因表达分析显示幼虫、未免疫成虫和免疫挑战成虫中表达的 AMPs 谱存在显著差异，这为发育阶段之间的稳定状态差异以及免疫诱导后发生的复杂转录重编程提供了深入了解。我们的数据深入了解了植食性昆虫所使用的适应机制，并可能导致开发出更有效的昆虫防治策略。
