College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.
Department of Entomology, Kansas State University, Manhattan, KS 66506, USA.
Pestic Biochem Physiol. 2018 Jul;149:143-148. doi: 10.1016/j.pestbp.2018.06.005. Epub 2018 Jun 15.
There has been a considerable growth in interest to use RNA interference (RNAi) as a novel insect pest management strategy in the past 10 years. However, there has been virtually no information on insect population variations in response to double-stranded RNA (dsRNA) molecules. The objective of this study was to generate baseline susceptibilities of the red flour beetle (Tribolium castaneum) to an insecticidal dsRNA targeting vacuolar H-ATPase subunit E gene (dsvATPaseE), and correlate the susceptibility data with sequence and expression variations of the target gene (vATPaseE), expression variations of the RNAi core genes, and overall genetic differences among a laboratory strain and seven geographical field populations of T. castaneum collected in China. Our results showed limited variations in the LD values of dsvATPaseE, which ranged from 0.10 to 0.29 ng/larva among the laboratory strain and the seven field populations. Considering the overlapping of the 95% confidence intervals of their LD values, there were no significant differences among the laboratory strain and field populations. We also found limited sequence polymorphisms and low frequencies of the polymorphisms of vATPaseE, and limited variations (<2-fold) of the endogenous expression of vATPaseE among the laboratory strain and field populations. However, we found considerable genetic variations among the individuals within each field population for most of eight loci and moderate to large genetic variations among the field populations. These results demonstrated that although the genetic variabilities were considerable among these field populations, the efficiency of RNAi targeting vATPaseE was highly consistent in T. castaneum. Our study provides work frames of resistance risk assessment for RNAi-based insect pest management programs.
在过去的 10 年中，人们对利用 RNA 干扰 (RNAi) 作为一种新的昆虫防治策略产生了浓厚的兴趣。然而，几乎没有关于昆虫种群对双链 RNA (dsRNA) 分子反应的变化的信息。本研究的目的是确定红粉甲虫 (Tribolium castaneum) 对靶向液泡 H+-ATP 酶亚基 E 基因 (dsvATPaseE) 的杀虫 dsRNA 的基本敏感性，并将敏感性数据与目标基因 (vATPaseE) 的序列和表达变化、RNAi 核心基因的表达变化以及在中国收集的实验室品系和七个地理野外种群之间的整体遗传差异相关联。我们的结果表明，dsvATPaseE 的 LD 值变化有限，实验室品系和七个野外种群之间的 LD 值范围为 0.10 至 0.29ng/幼虫。考虑到它们 LD 值的 95%置信区间重叠，实验室品系和野外种群之间没有显著差异。我们还发现 vATPaseE 的序列多态性有限，多态性频率低，实验室品系和野外种群中 vATPaseE 的内源性表达变化有限（<2 倍）。然而，我们发现，在大多数 8 个基因座内，每个野外种群中的个体之间存在相当大的遗传变异，而野外种群之间存在中等至较大的遗传变异。这些结果表明，尽管这些野外种群之间存在相当大的遗传变异性，但 vATPaseE 靶向 RNAi 的效率在红粉甲虫中高度一致。我们的研究为基于 RNAi 的昆虫防治计划的抗风险评估提供了工作框架。
