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The P\u2010element somatic inhibitor (PSI) protein, an exon splicing silencer that promotes male\u2010specific splicing of <jats:italic>dsx<\/jats:italic>, plays a critical role in male sexual differentiation and development. The functions of <jats:italic>PSI<\/jats:italic> have been characterized in the lepidopteran model species <jats:italic>Bombyx mori<\/jats:italic>. However, the molecular mechanism and functions of <jats:italic>PSI<\/jats:italic> in <jats:italic>Plutella xylostella<\/jats:italic>, a worldwide agricultural pest and taxonomically basal species, are still unknown.<\/jats:p><\/jats:sec><jats:sec><jats:title>RESULTS<\/jats:title><jats:p>Here we identified <jats:italic>PxPSI<\/jats:italic> transcripts and analyzed their spatiotemporal expression pattern in <jats:italic>P. xylostella<\/jats:italic>. Multiple sequence alignment revealed that PxPSI contains four KH domains and is highly conserved in lepidopterans. We used the CRISPR\u2013Cas9 system to generate mutations of the <jats:italic>PxPSI<\/jats:italic> genomic locus. Disruptions of <jats:italic>PxPSI<\/jats:italic> caused male\u2010specific defects in internal and external genitals. In addition, we detected female\u2010specific <jats:italic>Pxdsx<\/jats:italic> transcripts in <jats:italic>PxPSI<\/jats:italic> male mutants. Mutations also caused changes in expression of several sex\u2010biased genes and induced male sterility.<\/jats:p><\/jats:sec><jats:sec><jats:title>CONCLUSION<\/jats:title><jats:p>Our study demonstrates that <jats:italic>PxPSI<\/jats:italic> plays a key role in male sex determination in <jats:italic>P. xylostella<\/jats:italic> and suggests a potential molecular target for genetic\u2010based pest management in lepidopteran pests. \u00a9 2021 Society of Chemical Industry<\/jats:p><\/jats:sec>","DOI":"10.1002\/ps.6413","type":"journal-article","created":{"date-parts":[[2021,4,11]],"date-time":"2021-04-11T22:44:36Z","timestamp":1618181076000},"page":"3588-3596","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Mutation of <scp>P<\/scp>\u2010element somatic inhibitor induces male sterility in the diamondback moth, <i>Plutella xylostella<\/i>"],"prefix":"10.1002","volume":"77","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1903-1088","authenticated-orcid":false,"given":"Yaohui","family":"Wang","sequence":"first","affiliation":[{"name":"Hubei Key Laboratory of Insect Resource Application and Sustainable Pest Control, College of Plant Science &amp; Technology Huazhong Agricultural University  Wuhan China"},{"name":"CAS Key Laboratory of Insect Developmental and Evolutionary Biology CAS Center for Excellence in Molecular Plant Sciences\/Institute of Plant Physiology and Ecology  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xia","family":"Xu","sequence":"additional","affiliation":[{"name":"CAS Key Laboratory of Insect Developmental and Evolutionary Biology CAS Center for Excellence in Molecular Plant Sciences\/Institute of Plant Physiology and Ecology  Shanghai 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