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Deep orange gene editing triggers temperature-sensitive lethal phenotypes in Ceratitis capitata.

Germano Sollazzo^1,2,3, Katerina Nikolouli¹, Georgia Gouvi^1,4,3

¹Insect Pest Control Laboratory, Joint FAO/IAEA Centre of Nuclear Techniques in Food and Agriculture, Friedensstrasse 1, Seibersdorf, 2444, Austria.

BMC Biotechnology

|February 2, 2024

Summary

Researchers identified the deep orange gene in the Mediterranean fruit fly as a candidate for developing temperature-sensitive lethal (tsl) traits. Gene editing successfully created tsl mutant strains, offering a new tool for creating genetic sexing strains (GSS) in pest control.

Keywords:

Mediterranean fruit fly Sterile insect technique Temperature sensitivity Tephritidae White pupae

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Area of Science:

Entomology
Genetics
Pest Management

Background:

The Mediterranean fruit fly (Ceratitis capitata) is a major agricultural pest.
Area-wide integrated pest management (AW-IPM) and sterile insect technique (SIT) are key control strategies.
Genetic sexing strains (GSS) improve SIT efficiency by enabling male-only releases.

Purpose of the Study:

To investigate the deep orange (Ccdor) gene as a candidate for temperature-sensitive lethal (tsl) phenotypes.
To develop a generic approach for constructing GSS using selectable markers.
To identify and characterize mutations in Ccdor for GSS development.

Main Methods:

Cytogenetics, genomics, and bioinformatics were used to locate and characterize the Ccdor gene.
CRISPR/Cas9 gene editing (NHEJ and HDR) was employed to induce mutations in Ccdor.
Mutant strains were evaluated for tsl phenotypes and fitness costs at different temperatures.

Main Results:

The Ccdor gene was localized to chromosome 5 in the putative tsl region.
CRISPR/Cas9 gene editing resulted in Ccdor mutant strains exhibiting tsl phenotypes.
The developed strains showed minimal fitness costs at standard rearing temperatures (25°C) and complete lethality at elevated temperatures (36°C).

Conclusions:

Gene editing of the deep orange gene successfully generated temperature-sensitive lethal mutant strains in Ceratitis capitata.
These findings suggest that the conserved deep orange gene is a viable target for developing novel GSS.
This approach holds potential for creating GSS in other insect pests and disease vectors.