{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:29:03Z","timestamp":1785403743642,"version":"3.56.0"},"reference-count":19,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,7,15]],"date-time":"2026-07-15T00:00:00Z","timestamp":1784073600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"JSPS KAKENHI","award":["15K07805"],"award-info":[{"award-number":["15K07805"]}]},{"name":"JSPS KAKENHI","award":["15K01814"],"award-info":[{"award-number":["15K01814"]}]},{"name":"Joint Usage\/Research Center for Developmental Medicine, Institute of Molecular Embryology and Genetics, Kumamoto University"},{"id":[{"id":"https:\/\/ror.org\/02cgss904","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Digger wasps are solitary apoid wasps that excavate ground nests and use venom to paralyze insect prey. Recent phylogenetic analyses suggest that digger wasps belong to an ancient lineage of aculeate Hymenoptera and share an evolutionary ancestry with other venomous hymenopterans, including bees and ants. Although the venoms of ants, bees, and social wasps have been extensively studied, those of digger wasps remain poorly characterized in terms of their composition, molecular diversity, and biological activity. In this study, we investigated the venom of the digger wasp Cerceris japonica using integrated transcriptomic and proteomic analyses. We identified 19 toxin-like proteins and peptides, 14 non-toxin-associated components, and 11 novel peptides and proteins with no detectable similarity to known peptides and proteins. Among these, peptide Cj 2 exhibited insecticidal activity but showed no antimicrobial, hemolytic, or nicotinic acetylcholine receptor-modulating activities. These findings provide new insights into the molecular diversity and biological activities of digger wasp venom, expand our understanding of venom evolution in venomous hymenopterans, and serve as a framework for elucidating the conserved and lineage-specific features of hymenopteran venoms.<\/jats:p>","DOI":"10.3390\/toxins18070307","type":"journal-article","created":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T03:34:23Z","timestamp":1784172863000},"page":"307","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Integrated Venom Gland Transcriptomic and Venom Proteomic Analyses of the Digger Wasp Cerceris japonica"],"prefix":"10.3390","volume":"18","author":[{"given":"Kohei","family":"Kazuma","sequence":"first","affiliation":[{"name":"General Research Institute, Okinawa Churashima Foundation, 888 Aza Ishikawa, Motobu-cho, Kunigami-gun, Okinawa 905-0206, Okinawa, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naoki","family":"Tani","sequence":"additional","affiliation":[{"name":"Liaison Laboratory Research Promotion Center, Institute of Molecular Embryology and Genetics, Kumamoto University, 2-2-1 Honjo, Chuo-ku, Kumamoto 860-0811, Kumamoto, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katsuhiro","family":"Konno","sequence":"additional","affiliation":[{"name":"Graduate School of Agricultural Science, Tohoku University, 468-1 Aramaki Aza Aoba, Aoba-ku, Sendai 980-8572, Miyagi, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Migaku","family":"Kawaguchi","sequence":"additional","affiliation":[{"name":"Research Institute for Material and Chemical Measurement, National Metrology Institute of Japan (NMIJ), National Institute of Advanced Science and Technology (AIST), 1-1-1 Umezono, Tsukuba 305-8563, Ibaraki, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hidetoshi","family":"Inagaki","sequence":"additional","affiliation":[{"name":"Molecular Biosystems Research Institute, National Institute of Advanced Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566, Ibaraki, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Konno, K., Kazuma, K., and Nihei, K. (2016). Peptide toxins in solitary wasp venoms. Toxins, 8.","DOI":"10.3390\/toxins8040114"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Yoon, K.A., Kim, K., Kim, W.-J., Bang, W.Y., Ahn, N.-H., Bae, C.-H., Yeo, J.-H., and Lee, S.H. (2020). Characterization of venom components and their phylogenetic properties in some aculeate bumblebees and wasps. Toxins, 12.","DOI":"10.3390\/toxins12010047"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.toxicon.2019.07.012","article-title":"Analysis of venom sac constituents from the solitary aculeate wasp Cerceris rybyensis","volume":"169","author":"Kote","year":"2019","journal-title":"Toxicon"},{"key":"ref_4","first-page":"146","article-title":"Descriptions of new Hymenoptera from Japan. II","volume":"12","author":"Ashmead","year":"1904","journal-title":"J. N. Y. Entomol. Soc."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Tani, N., Kazuma, K., Ohtsuka, Y., Shigeri, Y., Masuko, K., Konno, K., and Inagaki, H. (2019). Mass spectrometry analysis and biological characterization of the predatory ant Odontomachus monticola venom and venom sac components. Toxins, 11.","DOI":"10.3390\/toxins11010050"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"945","DOI":"10.1515\/hsz-2019-0397","article-title":"Comprehensive analysis of peptides and low molecular weight components of the giant ant Dinoponera quadriceps venom","volume":"401","author":"Dodou","year":"2020","journal-title":"Biol. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Kim, G., and Hwang, U.W. (2023). The population genetic structure and phylogeographic dispersal of Nodularia breviconcha in the Korean Peninsula based on COI and 16S rRNA genes. PLoS ONE, 18.","DOI":"10.1371\/journal.pone.0288518"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Peiren, N., de Graaf, D.C., Vanrobaeys, F., Danneels, E.L., Devreese, B., Van Beeumen, J., and Jacobs, F.J. (2013). Proteome and phosphoproteome analysis of honeybee (Apis mellifera) venom collected from electrical stimulation and manual extraction of the venom gland. BMC Genom., 14.","DOI":"10.1186\/1471-2164-14-766"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"61","DOI":"10.7601\/mez.34.61","article-title":"Pattern analysis of biologically active amines in some hymenopteran venoms by high-performance liquid chromatography","volume":"34","author":"Nakajima","year":"1983","journal-title":"Jpn. J. Sanit. Zool."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1048","DOI":"10.1016\/j.bbrc.2005.03.087","article-title":"Molecular components and toxicity of the venom of the solitary wasp Anoplius samariensis","volume":"330","author":"Hisada","year":"2005","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Alberto-Silva, C., Vieira Portaro, F.C., Kodama, R.T., Pantale\u00e3o, H.Q., Inagaki, H., Nihei, K.I., and Konno, K. (2021). Comprehensive analysis and biological characterization of venom components from solitary scoliid wasp Campsomeriella annulata annulata. Toxins, 13.","DOI":"10.3390\/toxins13120885"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/S0022-3565(25)23227-X","article-title":"Allosteric inhibition of nicotinic acetylcholine receptors of vertebrates and insects by philanthotoxin","volume":"249","author":"Rozental","year":"1989","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1007\/s13659-015-0069-z","article-title":"A Novel Trypsin Inhibitor-Like Cysteine-Rich Peptide from the Frog Lepidobatrachus laevis Containing Proteinase-Inhibiting Activity","volume":"5","author":"Wang","year":"2015","journal-title":"Nat. Prod. Bioprospect."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Chen, Z., Wang, B., Hu, J., Yang, W., Cao, Z., Zhuo, R., Li, W., and Wu, Y. (2013). SjAPI, the First Functionally Characterized Ascaris-Type Protease Inhibitor from Animal Venoms. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0057529"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.jinsphys.2019.04.009","article-title":"A trypsin inhibitor-like protein secreted by Cotesia vestalis teratocytes inhibits hemolymph prophenoloxidase activation of Plutella xylostella","volume":"116","author":"Gu","year":"2019","journal-title":"J. Insect Physiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"msae263","DOI":"10.1093\/molbev\/msae263","article-title":"MEGA12: Molecular Evolutionary Genetic Analysis version 12 for adaptive and green computing","volume":"41","author":"Kumar","year":"2024","journal-title":"Mol. Biol. Evol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2114","DOI":"10.1093\/bioinformatics\/btu170","article-title":"Trimmomatic: A flexible trimmer for Illumina sequence data","volume":"30","author":"Bolger","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"644","DOI":"10.1038\/nbt.1883","article-title":"Full-length transcriptome assembly from RNA-Seq data without a reference genome","volume":"29","author":"Grabherr","year":"2011","journal-title":"Nat. Biotechnol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1080\/09168451.2015.1065169","article-title":"Pr-SNTX, a short-chain three-finger toxin from Papuan pygmy mulga snake, is an antagonist of muscle-type nicotinic acetylcholine receptor (\u03b12\u03b2\u03b4\u03b5)","volume":"80","author":"Yamauchi","year":"2016","journal-title":"Biosci. Biotechnol. Biochem."}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/18\/7\/307\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T03:46:21Z","timestamp":1784173581000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/18\/7\/307"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,15]]},"references-count":19,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["toxins18070307"],"URL":"https:\/\/doi.org\/10.3390\/toxins18070307","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,15]]}}}