College of Plant Protection, Nanjing Agricultural University, 210095, Nanjing, China.
Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, 1017, Denmark.
Nat Commun. 2018 Nov 16;9(1):4820. doi: 10.1038/s41467-018-07226-6.
The cotton bollworm Helicoverpa armigera, is one of the world's major pest of agriculture, feeding on over 300 hosts in 68 plant families. Resistance cases to most insecticide classes have been reported for this insect. Management of this pest in agroecosystems relies on a better understanding of how it copes with phytochemical or synthetic toxins. We have used genome editing to knock out a cluster of nine P450 genes and show that this significantly reduces the survival rate of the insect when exposed to two classes of host plant chemicals and two classes of insecticides. Functional expression of all members of this gene cluster identified the P450 enzymes capable of metabolism of these xenobiotics. The CRISPR-Cas9-based reverse genetics approach in conjunction with in vitro metabolism can rapidly identify the contributions of insect P450s in xenobiotic detoxification and serve to identify candidate genes for insecticide resistance.
棉铃虫(Helicoverpa armigera)是世界农业的主要害虫之一,以 68 科 300 多种植物为食。该昆虫对大多数杀虫剂类别都产生了抗药性。在农业生态系统中,对这种害虫的管理依赖于更好地了解它如何应对植物化学物质或合成毒素。我们利用基因组编辑敲除了一组 9 个 P450 基因,结果表明,当昆虫暴露于两类植物源化学物质和两类杀虫剂时,这一基因簇的存在显著降低了昆虫的存活率。对该基因簇所有成员的功能表达鉴定出了能够代谢这些外来化合物的 P450 酶。基于 CRISPR-Cas9 的反向遗传学方法与体外代谢相结合,可以快速鉴定出昆虫 P450 在异生物质解毒中的作用,并有助于鉴定候选的抗杀虫剂基因。