{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T12:58:30Z","timestamp":1790427510760,"version":"4.1.0"},"reference-count":24,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:00:00Z","timestamp":1788393600000},"content-version":"vor","delay-in-days":2,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100007565","name":"Citrus Research and Development Foundation","doi-asserted-by":"publisher","award":["CRDF23\u2010040"],"award-info":[{"award-number":["CRDF23\u2010040"]}],"id":[{"id":"10.13039\/100007565","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Arch Insect Biochem Physiol"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    The citrus root weevil,\n                    <jats:italic>Diaprepes abbreviatus<\/jats:italic>\n                    , is an economically important pest of citrus and ornamental crops whose subterranean larval feeding damages roots and predisposes plants to secondary pathogen infection. Development of efficient RNA interference (RNAi) delivery methods for early larval stages is essential for functional genomics studies and the evaluation of RNAi\u2010based pest management strategies. In this study, we developed a droplet\u2010based feeding assay for oral delivery of double\u2010stranded RNA (dsRNA) to neonates of\n                    <jats:italic>D. abbreviatus<\/jats:italic>\n                    using\n                    <jats:italic>chitin synthase 2<\/jats:italic>\n                    (\n                    <jats:italic>DaCHS2<\/jats:italic>\n                    ) as a model RNAi target to validate the assay. Feeding solutions containing dsRNA were supplemented with sucrose and bromophenol blue dye, with bromophenol blue used to visually confirm ingestion. Across three independent biological replicates, all neonates exposed to\n                    <jats:italic>DaCHS2<\/jats:italic>\n                    \u2010dsRNA,\n                    <jats:italic>GFP<\/jats:italic>\n                    \u2010dsRNA, and water control droplets were confirmed to have ingested the feeding solution (45\/45 neonates per treatment; 100% feeding success). Oral delivery of dsRNA targeting\n                    <jats:italic>DaCHS2<\/jats:italic>\n                    reduced transcript abundance and was associated with developmental abnormalities and mortality, including incomplete molting, abnormal pigmentation, cuticular deformities, defective pupation, and malformed adults. Regression analysis demonstrated moderate and significant relationship between dsRNA concentration and neonate mortality and developmental abnormalities. RT\u2010qPCR further confirmed reduced\n                    <jats:italic>DaCHS2<\/jats:italic>\n                    transcript abundance following oral dsRNA exposure. The developed assay provides a simple, reproducible, and minimally invasive proof\u2010of\u2010concept platform for oral dsRNA delivery to\n                    <jats:italic>D. abbreviatus<\/jats:italic>\n                    neonates. The assay requires only small dsRNA volumes, provides visual confirmation of ingestion, and may facilitate laboratory\u2010based screening of additional RNAi target genes in\n                    <jats:italic>D. abbreviatus<\/jats:italic>\n                    and other coleopteran pests.\n                  <\/jats:p>","DOI":"10.1002\/arch.70213","type":"journal-article","created":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T05:30:55Z","timestamp":1788499855000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of a Droplet\u2010Based RNA Interference Feeding Assay for Neonates of the Citrus Root Weevil\n                    <i>Diaprepes abbreviatus<\/i>"],"prefix":"10.1002","volume":"123","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5422-6695","authenticated-orcid":false,"given":"Tejbhan","family":"Saini","sequence":"first","affiliation":[{"name":"Department of Plant Pathology, Citrus Research and Education Center University of Florida Lake Alfred Florida USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3895-6943","authenticated-orcid":false,"given":"Nabil","family":"Killiny","sequence":"additional","affiliation":[{"name":"Department of Plant Pathology, Citrus Research and Education Center University of Florida Lake Alfred Florida USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,3]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1653\/024.092.0130"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pestbp.2015.04.004"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep38836"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10340-020-01197-8"},{"key":"e_1_2_10_6_1","doi-asserted-by":"crossref","unstructured":"Duncan L. W. L.Stelinski andL. M.Diepenbrock2025.2025\u20132026 Florida Citrus Production Guide: Citrus Root Weevils: CPG ch. 27 CG006\/ENY\u2010611 rev. 8\/2025. EDIS.https:\/\/doi.org\/10.32473\/edis-cg006-2023.","DOI":"10.32473\/edis-cg006-2023"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/1365-2664.12111"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pestbp.2021.104934"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0110536"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1006\/meth.2001.1262"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1186\/S12864-025-11880-8"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijerph8062265"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.2174\/1381612826666200523175409"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.3390\/insects13010040"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/arch.70153"},{"key":"e_1_2_10_16_1","volume-title":"Manual for Field Trials in Plant Protection","author":"Puntener W.","year":"1981"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-08813-1"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-07605-x"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/ps.71019"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.7150\/ijbs.14464"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11270-015-2686-x"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.est.0c04187"},{"key":"e_1_2_10_23_1","doi-asserted-by":"crossref","unstructured":"Weissling T. J. J. E.Pe\u00f1a R. M.Giblin\u2010Davis andJ. L.KnappJr.1998.Diaprepes Root Weevil Diaprepes abbreviatus (Linnaeus) (Insecta: Coleoptera: Curculionidae): EENY\u2010024\/IN151 2\/1998. EDIS.https:\/\/doi.org\/10.32473\/edis-in151-1998.","DOI":"10.32473\/edis-in151-1998"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13071-021-04741-2"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.3390\/f16060922"}],"container-title":["Archives of Insect Biochemistry and Physiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/arch.70213","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/arch.70213","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/arch.70213","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T12:12:38Z","timestamp":1790424758000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/arch.70213"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9]]},"references-count":24,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2026,9]]}},"alternative-id":["10.1002\/arch.70213"],"URL":"https:\/\/doi.org\/10.1002\/arch.70213","archive":["Portico"],"relation":{},"ISSN":["0739-4462","1520-6327"],"issn-type":[{"value":"0739-4462","type":"print"},{"value":"1520-6327","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9]]},"assertion":[{"value":"2026-07-17","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-08-22","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-03","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70213"}}