Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Competitive Sperm-marked Beetles for Monitoring Approaches in Genetic Biocontrol and Studies in Reproductive Biology

Version 1 : Received: 31 August 2022 / Approved: 16 September 2022 / Online: 16 September 2022 (03:02:52 CEST)

A peer-reviewed article of this Preprint also exists.

Isah, M.D.; Atika, B.; Dippel, S.; Ahmed, H.M.M.; Wimmer, E.A. Competitive Sperm-Marked Beetles for Monitoring Approaches in Genetic Biocontrol and Studies in Reproductive Biology. Int. J. Mol. Sci. 2022, 23, 12594. Isah, M.D.; Atika, B.; Dippel, S.; Ahmed, H.M.M.; Wimmer, E.A. Competitive Sperm-Marked Beetles for Monitoring Approaches in Genetic Biocontrol and Studies in Reproductive Biology. Int. J. Mol. Sci. 2022, 23, 12594.

Abstract

Sperm marking represents a valuable monitoring tool for genetic pest control strategies such as the Sterile Insect Technique, but also provides a key tool for reproductive biology studies. Sperm-marked lines can be generated by introducing transgenes that mediate the expression of fluorescent proteins during spermatogenesis. Homozygous lines established by transgenesis approaches are going through a genetic bottleneck that can lead to reduced fitness. Transgenic SIT approaches have mostly focused on Dipteran and Lepidopteran pests so far. With this study, we provide sperm-marked lines for the Coleopteran pest model organism, the red flour beetle Tribolium castaneum, based on the β2-tubulin promoter/enhancer driving red (DsRed) or green (EGFP) fluorescence. The obtained lines are reasonably competitive and were thus used for studies on reproductive biology confriming the phenomenon of ‘last male sperm precedence’ and that the spermathecae are deployed for long term sperm storage enabling the use of sperm from first matings even after secondary matings for a long period of time. The homozygosity and competiveness of the lines will enable future studies to analyze the controlled process of sperm movement into the long time storage organ as part of a post-mating cryptic female choice mechanism of this extremely promiscuous species.

Keywords

Insect Biotechnology; molecular entomology; pest management; Sterile Insect Technique; sperm storage; transgenesis; Tribolium castaneum

Subject

Biology and Life Sciences, Insect Science

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