Abstract
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.
Highlights
The cotton bollworm Helicoverpa armigera, is one of the world’s major pest of agriculture, feeding on over 300 hosts in 68 plant families
We recovered a homozygous line with the large deletion from one of the 18 single-pair families made between moths of G0 and the background SCD strain, suggesting that at least 2.8% (1/36) of the CYP6AE cluster deletion allele was induced and transmitted to the generation with the CRISPR-Cas[9] system in the current study
This demonstrates that this system can be useful in P450 monooxygenases (P450s) cluster knockout for H. armigera and for other insects in which the CRISPR-Cas[9] system is effective
Summary
The cotton bollworm Helicoverpa armigera, is one of the world’s major pest of agriculture, feeding on over 300 hosts in 68 plant families. 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. The CRISPR-Cas9based 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. The presence of multiple, closely related CYP genes in the genomes of pest insects presents a challenge to the functional identification of the genes that are important in adaptation to plant chemicals and detoxification of insecticides.
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