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

• Published in: Pest management science Vol. 80; no. 3; pp. 1219 - 1227
• Main Authors: Bai, Xue, Yu, Kaili, Xiong, Shijiao, Chen, Jin, Yang, Yi, Ye, Xinhai, Yao, Hongwei, Wang, Fang, Fang, Qi, Song, Qisheng, Ye, Gongyin
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01.03.2024; Wiley Subscription Services, Inc
• Subjects: Animals; Biological control; cocoon; Cocoons; CRISPR; CRISPR-Cas Systems; CRISPR/Cas9; Deoxyribonucleic acid; DNA; Ectoparasites; Editing; Embryos; Female; Genetic modification; Genome editing; Genomes; Genomics; Genotypes; Habrobracon hebetor; Mutagenesis; Mutants; non‐invasive; Offspring; Parameters; parasitic wasps; Pest control; Pests; Phenotypes; RNA, Guide, CRISPR-Cas Systems; Wasps - metabolism; White gene; genome editing; non-invasive; CRISPR/Cas9; parasitic wasps; cocoon
• ISSN: 1526-498X; 1526-4998
• DOI: 10.1002/ps.7851

BACKGROUND The 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. RESULTS To 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. CONCLUSION Our 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. Habrobracon hebetor white in the parasitoid wasp has been disrupted through CRISPR/Cas9, exhibiting reduced female rate and extended longevity in mutants. The cocoons have been utilized as a non‐invasive source for extracting genomic DNA, enabling non‐destructive genotyping.