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鉴定高效靶向基因用于 RNAi 介导控制灰绿白蜡虫,Agrilus planipennis。

Identification of highly effective target genes for RNAi-mediated control of emerald ash borer, Agrilus planipennis.

机构信息

University of Kentucky, Department of Entomology, Lexington, 40546, USA.

USDA ARS Beneficial Insects Introduction Research Unit, Newark, Delaware, USA.

出版信息

Sci Rep. 2018 Mar 22;8(1):5020. doi: 10.1038/s41598-018-23216-6.

Abstract

Recent study has shown that RNA interference (RNAi) is efficient in emerald ash borer (EAB), Agrilus planipennis, and that ingestion of double-stranded RNA (dsRNA) targeting specific genes causes gene silencing and mortality in neonates. Here, we report on the identification of highly effective target genes for RNAi-mediated control of EAB. We screened 13 candidate genes in neonate larvae and selected the most effective target genes for further investigation, including their effect on EAB adults and on a non-target organism, Tribolium castaneum. The two most efficient target genes selected, hsp (heat shock 70-kDa protein cognate 3) and shi (shibire), caused up to 90% mortality of larvae and adults. In EAB eggs, larvae, and adults, the hsp is expressed at higher levels when compared to that of shi. Ingestion of dsHSP and dsSHI caused mortality in both neonate larvae and adults. Administration of a mixture of both dsRNAs worked better than either dsRNA by itself. In contrast, injection of EAB.dsHSP and EAB.dsSHI did not cause mortality in T. castaneum. Thus, the two genes identified cause high mortality in the EAB with no apparent phenotype effects in a non-target organism, the red flour beetle, and could be used in RNAi-mediated control of this invasive pest.

摘要

最近的研究表明,RNA 干扰(RNAi)在翡翠灰螟(EAB)、榆绿天牛(Agrilus planipennis)中非常有效,摄入靶向特定基因的双链 RNA(dsRNA)会导致幼虫基因沉默和死亡。在这里,我们报告了用于 EAB 的 RNAi 介导的控制的高度有效的靶基因的鉴定。我们在幼虫期幼虫中筛选了 13 个候选基因,并选择了最有效的靶基因进行进一步研究,包括它们对 EAB 成虫和非靶标生物赤拟谷盗(Tribolium castaneum)的影响。选择的两个最有效的靶基因 hsp(热休克 70kDa 蛋白同源物 3)和 shi(shibire)导致幼虫和成虫的死亡率高达 90%。在 EAB 卵、幼虫和成虫中,与 shi 相比,hsp 的表达水平更高。dsHSP 和 dsSHI 的摄入导致幼虫和成虫的死亡率。两种 dsRNA 的混合物的给药比单独的任何一种 dsRNA 效果更好。相比之下,EAB.dsHSP 和 EAB.dsSHI 的注射不会导致赤拟谷盗的死亡。因此,这两个基因在非靶标生物——红粉甲虫中导致 EAB 高死亡率,而没有明显的表型影响,并且可以用于 RNAi 介导的这种入侵性害虫的控制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5624/5864839/b64f52f878b1/41598_2018_23216_Fig1_HTML.jpg

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