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Abstract
Recently, a novel sex‐determination system was identified in the silkworm (Bombyx mori) in which a piwi‐interacting RNA (piRNA) encoded on the female‐specific W chromosome silences a Z‐linked gene (Masculinizer) that would otherwise initiate male sex‐determination and dosage compensation. Masculinizer provides various opportunities for developing improved genetic pest management tools. A pest lepidopteran in which a genetic pest management system has been developed, but which would benefit greatly from such improved designs, is the diamondback moth, Plutella xylostella. However, Masculinizer has not yet been identified in this species. Here, focusing on the previously described ‘masculinizing’ domain of B. mori Masculinizer, we identify P. xylostella Masculinizer (PxyMasc). We show that PxyMasc is Z‐linked, regulates sex‐specific alternative splicing of doublesex and is necessary for male survival. Similar results in B. mori suggest this survival effect is possibly through failure to initiate male dosage compensation. The highly conserved function and location of this gene between these two distantly related lepidopterans suggests a deep role for Masculinizer in the sex‐determination systems of the Lepidoptera.
Highlights
Sex-determination systems are of fundamental biological interest and provide targets and components for genetic pest management systems, including gene drives (Burt, 2003; KaramiNejadRanjbar et al, 2018)
Masc is further involved in regulating dosage compensation of Z-linked genes in B. mori as short-interfering RNA-based disruption of Masc messenger RNA in eggs led to male-specific embryonic lethality
Unlike in B. mori, we are unable to sex diamondback moth (DBM) individuals at the egg stage but could test for this phenotype by assessing whether hatch rates of eggs injected with double-stranded RNA targeting the two putative Masc homologues diverged significantly from control injections (dsRNA against Anemonia majano Cyan (AmCyan) coding sequence (CDS)) – if all male embryos died owing to the treatment, the hatch rate would be half that of control injections
Summary
Sex-determination systems are of fundamental biological interest and provide targets and components for genetic pest management systems, including gene drives (Burt, 2003; KaramiNejadRanjbar et al, 2018). We reported a novel genetic pest management system that utilized a dsx splicing cassette from pink bollworm (Pectinophora gossypiella, PBW) to engineer female-specific lethality into DBM (Jin et al, 2013). This system was shown to work effectively in B. mori (Tan et al, 2013). Previous research on B. mori Masc identified the amino acid region between 304 and 310 aa and the two cysteine residues (cysteinecysteine domain) as being necessary for promoting male-specific splicing of dsx (Katsuma et al, 2015) This region shows a relatively high degree of homology across the rather divergent putative Masc homologues identified, albeit from a restricted phylogenetic range within the Macroheterocera and Papilionoidea. Identification of PxyMasc extends the conservation of this unique sexdetermination system across the Lepidoptera and provides a target both for further research into upstream DBM sex-determination components, as well as for building gene drive population suppression systems
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