Abstract
The sterile insect technique (SIT) is a highly effective biologically-based method for the population suppression of highly invasive insect pests of medical and agricultural importance. The efficacy of SIT could be significantly enhanced, however, by improved methods of male sterilization that avoid the fitness costs of irradiation. An alternative sterilization method is possible by gene-editing that targets genes essential for sperm maturation and motility, rendering them nonfunctional, similar to the CRISPR-Cas9 targeting of β2-tubulin in the genetic model system, Drosophila melanogaster. However, since genetic strategies for sterility are susceptible to breakdown or resistance in mass-reared populations, alternative targets for sterility are important for redundancy or strain replacement. Here we have identified and characterized the sequence and transcriptional expression of two genes in a Florida strain of Drosophila suzukii, that are cognates of the D. melanogaster spermatocyte-specific genes wampa and Prosalpha6T. Wampa encodes a coiled-coil dynein subunit required for axonemal assembly, and the proteasome subunit gene, Prosalpha6T, is required for spermatid individualization and nuclear maturation. The reading frames of these genes differed from their NCBI database entries derived from a D. suzukii California strain by 44 and 8 nucleotide substitutions/polymorphisms, respectively, though all substitutions were synonymous resulting in identical peptide sequences. Expression of both genes is predominant in the male testis, and they share similar transcriptional profiles in adult males with β2-tubulin. Their amino acid sequences are highly conserved in dipteran species, including pest species subject to SIT control, supporting their potential use in targeted male sterilization strategies.
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Data availability
The datasets generated by the current study are available from NCBI GenBank as noted in the text and the corresponding author on reasonable request.
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Acknowledgements
Grateful appreciation is extended to Richard Furlong and Shelley Olson for technical assistance with insect rearing and experimental procedures.
Funding
This project was supported by the Biotechnology Risk Assessment Research Grants Program Grant No. 2020-33522-3227 from the USDA National Institute of Food and Agriculture to A.M.H., and USDA-ARS Agreement 59-6036-1-002 and USDA National Institute of Food and Agriculture Hatch project FLA-ENY-005943 to D.A.H.
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QX, KT and AMH conceptualized the experimental approach and methodology; QX and KT performed experiments; QX, KT, DAH and AMH validated data and results; and AMH and DAH acquired funding and supervised research. All authors reviewed the manuscript.
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This is a biological, genetic and bioinformatic study of insect gene sequences and their expression using biochemical and in silico tools and resources on insect specimens that does not require ethical approval.
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Xia, Q., Tariq, K., Hahn, D.A. et al. Sequence and expression analysis of the spermatogenesis-specific gene cognates, wampa and Prosα6T, in Drosophila suzukii. Genetica 151, 215–223 (2023). https://doi.org/10.1007/s10709-023-00189-7
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DOI: https://doi.org/10.1007/s10709-023-00189-7