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This study aimed to investigate the expression and function of the nuclear hormone receptor 3 (<jats:italic>HR3<\/jats:italic>) in the 28\u2010spotted ladybeetle, <jats:italic>Henosepilachna vigintioctopunctata<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>RESULTS<\/jats:title><jats:p>Our findings revealed that <jats:italic>HvHR3<\/jats:italic> was expressed at all developmental stages, with the highest expression levels in the 1st instar larvae, 3rd instar larvae, and pupa. <jats:italic>HvHR3<\/jats:italic> silencing in both the 1st and 3rd instar larvae resulted in high mortality rates. Reduced <jats:italic>HvHR3<\/jats:italic> expression in the 3rd instar larvae led to developmental arrest and failure to molt, accompanied by a down\u2010regulation of 20E\u2010related genes. Furthermore, <jats:italic>HvHR3<\/jats:italic> silencing in the 3rd instar larvae resulted in a decrease in the 20E titer. Silencing <jats:italic>HvHR3<\/jats:italic> in adult females significantly impaired ovarian development, resulting in reduced ovary size and absence of mature oocytes. Additionally, the eggs produced by these females had partially ruptured shells and failed to maintain normal spindle shape.<\/jats:p><\/jats:sec><jats:sec><jats:title>CONCLUSION<\/jats:title><jats:p>These findings indicate that <jats:italic>HvHR3<\/jats:italic> is a critical regulator of development, molting, and reproduction, underscoring its potential as a molecular target for RNA interference\u2010based biopesticides aimed at controlling this pest. \u00a9 2025 Society of Chemical Industry.<\/jats:p><\/jats:sec>","DOI":"10.1002\/ps.8810","type":"journal-article","created":{"date-parts":[[2025,4,5]],"date-time":"2025-04-05T05:32:12Z","timestamp":1743831132000},"page":"4530-4539","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["<scp>RNAi<\/scp> assay in the 28\u2010spotted ladybeetle <i>Henosepilachna vigintioctopunctata<\/i> suggests nuclear receptor <scp><i>HR3<\/i><\/scp> as a potential molecular target for pest 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