CRISPR/Cas9‐mediated mutagenesis of the white gene in an ectoparasitic wasp, Habrobracon hebetor

Author:

Bai Xue1ORCID,Yu Kaili1,Xiong Shijiao1,Chen Jin1,Yang Yi1,Ye Xinhai1,Yao Hongwei1,Wang Fang1,Fang Qi1ORCID,Song Qisheng2,Ye Gongyin1ORCID

Affiliation:

1. State Key Laboratory of Rice Biology and Breeding, Ministry of Agricultural and Rural Affairs Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang Province, Institute of Insect Sciences, College of Agriculture and Biotechnology Zhejiang University Hangzhou China

2. Division of Plant Science and Technology College of Agriculture, Food and Natural Resources, University of Missouri Columbia MO USA

Abstract

AbstractBACKGROUNDThe ectoparasitic wasp Habrobracon hebetor (Hymenoptera, Braconidae) can parasitize various species of lepidopteran pests. To maximize its potential for biological control, it is necessary to investigate its gene function through genome engineering.RESULTSTo test the effectiveness of genome engineering system in H. hebetor, we injected the mixture of clustered regularly interspaced short palindromic repeats (CRISPR) ‐associated (Cas) 9 protein and single guide RNA(s) targeting gene white into embryos. The resulting mutants display a phenotype of eye pigment loss. The phenotype was caused by small indel and is heritable. Then, we compared some biological parameters between wildtype and mutant, and found there were no significant differences in other parameters except for the offspring female rate and adult longevity. In addition, cocoons could be used to extract genomic DNA for genotype during the gene editing process without causing unnecessary harm to H. hebetor.CONCLUSIONOur results demonstrate that the CRISPR/Cas9 system can be used for H. hebetor genome editing and it does not adversely affect biological parameters of the parasitoid wasps. We also provide a feasible non‐invasive genotype detection method using genomic DNA extracted from cocoons. Our study introduces a novel tool and method for studying gene function in H. hebetor, and may contribute to better application of H. hebetor in biocontrol. © 2023 Society of Chemical Industry.

Publisher

Wiley

Subject

Insect Science,Agronomy and Crop Science,General Medicine

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