Lu Yong, Chen Mengyao, Reding Katie, Pick Leslie
Department of Entomology, University of Maryland, College Park, MD 20742 USA.
Present Address: Department of Anesthesiology, Stony Brook Medicine, 101 Nicolls Rd, Stony Brook, NY 11794 USA.
Evodevo. 2017 Oct 18;8:15. doi: 10.1186/s13227-017-0078-6. eCollection 2017.
Hemiptera is a large clade of insects understudied in terms of developmental biology. , the Brown Marmorated Stink Bug (BMSB, referred to throughout as ), is an invasive hemipteran pest of the mid-Atlantic region of the USA that has rapidly spread to other regions in recent years, devastating a wide range of crops using a piercing and sucking mechanism. Its phylogenetic position, polyphagous habits, and rapid spread in the USA suggested that would be an ideal system to broaden our knowledge of developmental mechanisms in insects. We and others previously generated transcriptome sequences from different life stages of this insect. Here, we describe tools to examine gene expression patterns in whole-mount embryos and to test the response of to RNA interference (RNAi). We show that spatial and temporal patterns of gene expression in can be effectively monitored by both immunostaining and in situ hybridization. We also show that delivery of dsRNA to adult females knocks down gene function in offspring, using the homeotic gene (). Knockdown of - resulted in dramatic malformations of the mouthparts, demonstrating for the first time that RNAi is effective in this species. Our results suggest that, despite difficulties with long-term laboratory culture of , this species shows promise as a developmental system.
半翅目昆虫是发育生物学研究较少的一大类昆虫。褐边绿刺蛾（BMSB，以下简称“该昆虫”）是美国大西洋中部地区的一种入侵性半翅目害虫，近年来迅速蔓延至其他地区，通过刺吸机制对多种作物造成严重破坏。其系统发育位置、多食性习性以及在美国的快速传播表明，该昆虫将是一个理想的系统，有助于拓宽我们对昆虫发育机制的认识。我们和其他人之前已从该昆虫的不同生命阶段生成了转录组序列。在此，我们描述了用于检测整装胚胎中基因表达模式以及测试该昆虫对RNA干扰（RNAi）反应的工具。我们表明，通过免疫染色和原位杂交均可有效监测该昆虫基因表达的时空模式。我们还表明，将双链RNA（dsRNA）导入成年雌虫可敲低其后代的基因功能，以同源异型基因（）为例。敲低 - 会导致口器出现严重畸形，首次证明RNAi在该物种中有效。我们的结果表明，尽管该昆虫在长期实验室培养方面存在困难，但作为一个发育系统，该物种具有潜力。
