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To address this, we used mRNA-seq to isolate genes differentially expressed between ovary-active and ovary-inactive workers in the intermediately eusocial bumblebee Bombus terrestris. By comparisons with data from the advanced eusocial honeybee Apis mellifera, which shows reduced worker reproductivity, we characterized gene expression differences associated with change in worker reproductivity as a function of eusocial complexity. By comparisons with genes associated with queen-worker caste development in B. terrestris larvae, we tested the behavioral\u2013morphological caste homology hypothesis, which proposes co-option of genes influencing reproductive division of labor in adults in morphological caste evolution. We conducted comparisons having isolated genes expressed in B. terrestris worker-laid eggs to remove the potential confound caused by gene expression in eggs. Gene expression differences between the B. terrestris worker phenotypes were mainly in fat body and ovary, not brain. Many genes (86%) more highly expressed in ovary of ovary-active workers were also expressed in worker-laid eggs, confirming egg-expressed genes were potentially confounding. Comparisons across B. terrestris and A. mellifera, and with B. terrestris larvae, returned significant percentage overlaps in differentially expressed genes and\/or enriched Gene Ontology terms, suggesting conserved gene functions underpin worker reproductivity as it declines with increasing eusocial complexity and providing support for the behavioral\u2013morphological caste homology hypothesis. Therefore, within bees, both a degree of conserved gene use and gene co-option appear to underlie the molecular basis of worker reproductivity and morphological caste evolution.<\/jats:p>","DOI":"10.1093\/gbe\/evae269","type":"journal-article","created":{"date-parts":[[2024,12,12]],"date-time":"2024-12-12T08:05:22Z","timestamp":1733990722000},"source":"Crossref","is-referenced-by-count":3,"title":["Molecular Basis of Eusocial Complexity: The Case of Worker Reproductivity in Bees"],"prefix":"10.1093","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2214-9733","authenticated-orcid":false,"given":"David C","family":"Prince","sequence":"first","affiliation":[{"name":"School of Biological Sciences, University of East Anglia, Norwich Research Park , Norwich NR4 7TJ ,","place":["UK"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6416-052X","authenticated-orcid":false,"given":"Anders","family":"Wir\u00e9n","sequence":"additional","affiliation":[{"name":"School of Biological Sciences, University of East Anglia, Norwich Research Park , Norwich NR4 7TJ ,","place":["UK"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Timothy J","family":"Huggins","sequence":"additional","affiliation":[{"name":"School of Biological Sciences, University of East Anglia, Norwich Research Park , Norwich NR4 7TJ ,","place":["UK"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8619-8158","authenticated-orcid":false,"given":"David H","family":"Collins","sequence":"additional","affiliation":[{"name":"School of Biological Sciences, University of East Anglia, Norwich Research Park , Norwich NR4 7TJ ,","place":["UK"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1492-5429","authenticated-orcid":false,"given":"Tamas","family":"Dalmay","sequence":"additional","affiliation":[{"name":"School of Biological Sciences, University of East Anglia, Norwich Research Park , Norwich NR4 7TJ ,","place":["UK"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5891-8816","authenticated-orcid":false,"given":"Andrew F G","family":"Bourke","sequence":"additional","affiliation":[{"name":"School of Biological Sciences, University of East Anglia, Norwich Research Park , Norwich NR4 7TJ ,","place":["UK"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2024,12,12]]},"reference":[{"issue":"8","key":"2024122616224292400_evae269-B1","doi-asserted-by":"publisher","first-page":"400","DOI":"10.1007\/s00114-004-0547-3","article-title":"Does the queen win it all? 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