{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T03:55:42Z","timestamp":1788407742666,"version":"build-2803163510"},"reference-count":48,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2025,5,15]],"date-time":"2025-05-15T00:00:00Z","timestamp":1747267200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Tianshan Talents Cultivation Program","award":["2023TSYCCX0015"],"award-info":[{"award-number":["2023TSYCCX0015"]}]},{"name":"Tianshan Talents Cultivation Program","award":["CARS17"],"award-info":[{"award-number":["CARS17"]}]},{"name":"China Agriculture Research System","award":["2023TSYCCX0015"],"award-info":[{"award-number":["2023TSYCCX0015"]}]},{"name":"China Agriculture Research System","award":["CARS17"],"award-info":[{"award-number":["CARS17"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Insects"],"abstract":"<jats:p>The clover cutworm, Scotogramma trifolii Rottemberg (Lepidoptera: Noctuidae), is a globally distributed polyphagous pest causing significant economic losses to agricultural crops. RT-qPCR is a gold-standard technique for gene expression analysis, yet its accuracy depends critically on stable reference genes for data normalization. To address the lack of validated reference genes in S. trifolii, we evaluated six candidate genes (\u03b2-actin, RPL9, GAPDH, RPL10, EF1-\u03b1, and TUB) across four developmental stages (egg, larva, pupa, and adult) and six adult tissues (head, thorax, abdomen, wings, legs, and antennae) using geNorm, NormFinder, BestKeeper, and RefFinder algorithms. Stability analysis identified \u03b2-actin, RPL9, and GAPDH as the most reliable reference genes for developmental stage normalization, while RPL10, GAPDH, and TUB were validated for adult tissues. Functional validation using the odorant receptor gene StriOR20 revealed significant discrepancies in relative expression levels when normalized with unstable reference genes (TUB and RPL9), emphasizing the necessity of rigorous reference gene selection. This study establishes the first comprehensive reference gene panel for S. trifolii, providing a robust foundation for gene expression studies in this agriculturally important pest.<\/jats:p>","DOI":"10.3390\/insects16050527","type":"journal-article","created":{"date-parts":[[2025,5,15]],"date-time":"2025-05-15T07:46:49Z","timestamp":1747295209000},"page":"527","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Selection and Validation of Stable Reference Genes for RT-qPCR in Scotogramma trifolii (Lepidoptera: Noctuidae)"],"prefix":"10.3390","volume":"16","author":[{"given":"Anpei","family":"Yang","sequence":"first","affiliation":[{"name":"Institute of Plant Protection, Xinjiang Uygur Autonomous Region Academy of Agricultural Sciences, Urumqi 830091, China"},{"name":"Key Laboratory of Integrated Pest Management on Crops in Northwestern Oasis, Ministry of Agriculture and Rural Affairs, National Plant Protection Scientific Observation and Experiment Station of Korla, Urumqi 830091, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Institute of Plant Protection, Xinjiang Uygur Autonomous Region Academy of Agricultural Sciences, Urumqi 830091, China"},{"name":"Key Laboratory of Integrated Pest Management on Crops in Northwestern Oasis, Ministry of Agriculture and Rural Affairs, National Plant Protection Scientific Observation and Experiment Station of Korla, Urumqi 830091, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weiwei","family":"Bai","sequence":"additional","affiliation":[{"name":"Institute of Plant Protection, Xinjiang Uygur Autonomous Region Academy of Agricultural Sciences, Urumqi 830091, China"},{"name":"Key Laboratory of Integrated Pest Management on Crops in Northwestern Oasis, Ministry of Agriculture and Rural Affairs, National Plant Protection Scientific Observation and Experiment Station of Korla, Urumqi 830091, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruifeng","family":"Ding","sequence":"additional","affiliation":[{"name":"Institute of Plant Protection, Xinjiang Uygur Autonomous Region Academy of Agricultural Sciences, Urumqi 830091, China"},{"name":"Key Laboratory of Integrated Pest Management on Crops in Northwestern Oasis, Ministry of Agriculture and Rural Affairs, National Plant Protection Scientific Observation and Experiment Station of Korla, Urumqi 830091, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weipeng","family":"Li","sequence":"additional","affiliation":[{"name":"Institute of Plant Protection, Xinjiang Uygur Autonomous Region Academy of Agricultural Sciences, Urumqi 830091, China"},{"name":"Key Laboratory of Integrated Pest Management on Crops in Northwestern Oasis, Ministry of Agriculture and Rural Affairs, National Plant Protection Scientific Observation and Experiment Station of Korla, Urumqi 830091, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guangkuo","family":"Li","sequence":"additional","affiliation":[{"name":"Institute of Plant Protection, Xinjiang Uygur Autonomous Region Academy of Agricultural Sciences, Urumqi 830091, China"},{"name":"Key Laboratory of Integrated Pest Management on Crops in Northwestern Oasis, Ministry of Agriculture and Rural Affairs, National Plant Protection Scientific Observation and Experiment Station of Korla, Urumqi 830091, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/0022-2011(82)90035-0","article-title":"A new type of insect pathogen in larvae of the clover cutworm, Scotogramma trifolii","volume":"40","author":"Federici","year":"1982","journal-title":"J. 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