{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,9]],"date-time":"2026-04-09T18:25:51Z","timestamp":1775759151115,"version":"3.50.1"},"reference-count":42,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2026,4,9]],"date-time":"2026-04-09T00:00:00Z","timestamp":1775692800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key R&D Program of China","doi-asserted-by":"crossref","award":["2022YFD1600205"],"award-info":[{"award-number":["2022YFD1600205"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100004607","name":"Natural Science Foundation of Guangxi Province","doi-asserted-by":"publisher","award":["2024GXNSFAA010181"],"award-info":[{"award-number":["2024GXNSFAA010181"]}],"id":[{"id":"10.13039\/501100004607","id-type":"DOI","asserted-by":"publisher"}]},{"name":"China Agriculture Research System-Bee","award":["CARS-44-KXJ5"],"award-info":[{"award-number":["CARS-44-KXJ5"]}]},{"name":"Agricultural Science and Technology Innovation Program, Chinese Academy of Agricultural Sciences","award":["CAAS-ASTIP-2025-IAR"],"award-info":[{"award-number":["CAAS-ASTIP-2025-IAR"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    Drones are the males in the honey bee population that provide high quantity and quality sperm to mate with the queen and ensure colony reproduction. To understand spermatogenesis in the drone testis at the molecular level, reverse transcription quantitative PCR (RT-qPCR) is a standard and useful means to investigate the target genes. However, a lack of appropriate reference genes hinders the accurate normalization of target genes. To identify stable reference genes in the drone testis during the meiosis stages of honey bee, we assessed eight candidate genes using BestKeeper, delta-CT, GeNorm, NormFinder, and RefFinder tools. Moreover, the expression pattern of\n                    <jats:italic>stat92e<\/jats:italic>\n                    and\n                    <jats:italic>dicer1<\/jats:italic>\n                    was used to validate the stability of the selected reference genes. The results showed that the most stable reference gene in the testis during meiosis stages varied according to different algorithms, and\n                    <jats:italic>gapdh<\/jats:italic>\n                    was the most stable reference gene in the drone testis during meiosis stages when integrating these different algorithms. In addition,\n                    <jats:italic>gapdh<\/jats:italic>\n                    ,\n                    <jats:italic>rps18<\/jats:italic>\n                    , and the combination of\n                    <jats:italic>gapdh<\/jats:italic>\n                    &amp;\n                    <jats:italic>rps18<\/jats:italic>\n                    were optimal reference genes to normalize the expression of the target genes in testis during meiosis stages, as validated by the expression of\n                    <jats:italic>stat92e<\/jats:italic>\n                    and\n                    <jats:italic>dicer1<\/jats:italic>\n                    . Our findings provide a conducive foundation for the accurate quantification of gene expression levels in the testis during the drone meiosis stages of the honey bee.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0347110","type":"journal-article","created":{"date-parts":[[2026,4,9]],"date-time":"2026-04-09T17:30:32Z","timestamp":1775755832000},"page":"e0347110","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Reference gene selection for RT-qPCR normalization in the drone testis of honey bee (Apis cerana) during meiosis stages"],"prefix":"10.1371","volume":"21","author":[{"given":"Lifenfang","family":"Tao","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xinran","family":"Fu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chunxiu","family":"Pang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2628-0624","authenticated-orcid":true,"given":"Yanan","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5373-6108","authenticated-orcid":true,"given":"Jiaxing","family":"Huang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2026,4,9]]},"reference":[{"issue":"1608","key":"pone.0347110.ref001","first-page":"303","article-title":"Importance of pollinators in changing landscapes for world crops","volume":"274","author":"A-M Klein","year":"2007","journal-title":"Proc Biol Sci"},{"issue":"6","key":"pone.0347110.ref002","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1016\/j.tree.2010.01.007","article-title":"Global pollinator declines: trends, impacts and drivers","volume":"25","author":"SG Potts","year":"2010","journal-title":"Trends Ecol Evol"},{"issue":"8","key":"pone.0347110.ref003","doi-asserted-by":"crossref","first-page":"2381","DOI":"10.1111\/j.1365-294X.2004.02262.x","article-title":"Extreme queen-mating frequency and colony fission in African army ants","volume":"13","author":"DJC Kronauer","year":"2004","journal-title":"Mol Ecol"},{"issue":"1","key":"pone.0347110.ref004","doi-asserted-by":"crossref","first-page":"11","DOI":"10.3390\/insects10010011","article-title":"Reproductive Senescence in Drones of the Honey Bee (Apis mellifera)","volume":"10","author":"BN Metz","year":"2019","journal-title":"Insects"},{"issue":"7","key":"pone.0347110.ref005","doi-asserted-by":"crossref","first-page":"1581","DOI":"10.1016\/j.sjbs.2018.10.012","article-title":"Reproductive biology and morphology of Apis mellifera jemenitica (Apidae) queens and drones","volume":"26","author":"R Al-Sarhan","year":"2019","journal-title":"Saudi J Biol Sci"},{"issue":"3","key":"pone.0347110.ref006","first-page":"124","article-title":"Histopathological Findings in Testes from Apparently Healthy Drones of Apis mellifera ligustica","volume":"7","author":"K Power","year":"2020","journal-title":"Vet Sci"},{"issue":"4","key":"pone.0347110.ref007","first-page":"105","article-title":"Development and secretory nature of seminal vesicle during sexual maturation in Indian honeybee, Apis cerana indica F. 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