{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,9]],"date-time":"2026-04-09T02:48:00Z","timestamp":1775702880521,"version":"3.50.1"},"reference-count":70,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2024,1,15]],"date-time":"2024-01-15T00:00:00Z","timestamp":1705276800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Slovenian Research and Innovation Agency  (ARIS)","award":["J4-2543"],"award-info":[{"award-number":["J4-2543"]}]},{"name":"Slovenian Research and Innovation Agency  (ARIS)","award":["P4-0432"],"award-info":[{"award-number":["P4-0432"]}]},{"name":"Slovenian Research and Innovation Agency  (ARIS)","award":["P1-0184"],"award-info":[{"award-number":["P1-0184"]}]},{"name":"Slovenian Research and Innovation Agency  (ARIS)","award":["134356 SNN20"],"award-info":[{"award-number":["134356 SNN20"]}]},{"name":"National Research, Development and Innovation Office of Hungary (NKFIH)","award":["J4-2543"],"award-info":[{"award-number":["J4-2543"]}]},{"name":"National Research, Development and Innovation Office of Hungary (NKFIH)","award":["P4-0432"],"award-info":[{"award-number":["P4-0432"]}]},{"name":"National Research, Development and Innovation Office of Hungary (NKFIH)","award":["P1-0184"],"award-info":[{"award-number":["P1-0184"]}]},{"name":"National Research, Development and Innovation Office of Hungary (NKFIH)","award":["134356 SNN20"],"award-info":[{"award-number":["134356 SNN20"]}]},{"name":"core donors","award":["J4-2543"],"award-info":[{"award-number":["J4-2543"]}]},{"name":"core donors","award":["P4-0432"],"award-info":[{"award-number":["P4-0432"]}]},{"name":"core donors","award":["P1-0184"],"award-info":[{"award-number":["P1-0184"]}]},{"name":"core donors","award":["134356 SNN20"],"award-info":[{"award-number":["134356 SNN20"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Insects"],"abstract":"<jats:p>Certain soil insects, such as the root-damaging larvae of the maize pest Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae), are increasingly difficult to control because of recent bans of some insecticides. An alternative and safer approach may be the development of biopesticides based on entomotoxic defense proteins of higher fungi. Many of these potentially interesting proteins are protease inhibitors, and some have been shown to adversely affect insects. We examined the effects of the cysteine protease inhibitors macrocypin 1, 3, and 4 from Macrolepiota procera, clitocypin from Clitocybe nebularis, and cocaprin 1 and the serine protease inhibitor cospin 1 from Coprinopsis cinerea on D. v. virgifera. We confirmed the inhibition by mycocypins of the cysteine catalytic-type proteolytic activities in gut extracts of larvae and adults. The inhibition of pGlu-Phe-Leu-hydrolyzing activity was stronger than that of Z-Phe-Arg-hydrolyzing activity. Mycocypins and cospin resisted long-term proteolytic digestion, whereas cocaprin 1 was digested. Bioassays with overlaid artificial diet revealed no effects of proteins on neonatal mortality or stunting, and no effects on adult mortality. Immersion of eggs in protein solutions had little effect on egg hatching or mortality of hatching neonates. Microscopic analysis of the peritrophic matrix and apical surface of the midguts revealed the similarity between larvae of D. v. virgifera and the chrysomelid Leptinotarsa decemlineata, which are sensitive to these inhibitors. The resistance of D. v. virgifera to fungal protease inhibitors is likely due to effective adaptation of digestive enzyme expression to dietary protease inhibitors. We continue to study unique protein complexes of higher fungi for the development of new approaches to pest control.<\/jats:p>","DOI":"10.3390\/insects15010060","type":"journal-article","created":{"date-parts":[[2024,1,15]],"date-time":"2024-01-15T08:52:03Z","timestamp":1705308723000},"page":"60","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Diabrotica v. virgifera Seems Not Affected by Entomotoxic Protease Inhibitors from Higher Fungi"],"prefix":"10.3390","volume":"15","author":[{"given":"Stefan","family":"Toepfer","sequence":"first","affiliation":[{"name":"Department of Integrated Plant Protection, Plant Protection Institute, Hungarian University of Agriculture and Life Sciences (MATE), 2100 Godollo, Hungary"},{"name":"CABI, 2800 Delemont, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4238-2361","authenticated-orcid":false,"given":"Szabolcs","family":"Toth","sequence":"additional","affiliation":[{"name":"Department of Integrated Plant Protection, Plant Protection Institute, Hungarian University of Agriculture and Life Sciences (MATE), 2100 Godollo, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2336-0634","authenticated-orcid":false,"given":"Tanja","family":"Zupan","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Jo\u017eef Stefan Institute, 1000 Ljubljana, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Urban","family":"Bogataj","sequence":"additional","affiliation":[{"name":"Department of Biology, Biotechnical Faculty, University of Ljubljana, 1000 Ljubljana, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nada","family":"\u017dnidar\u0161i\u010d","sequence":"additional","affiliation":[{"name":"Department of Biology, Biotechnical Faculty, University of Ljubljana, 1000 Ljubljana, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3114-8289","authenticated-orcid":false,"given":"Marta","family":"Ladanyi","sequence":"additional","affiliation":[{"name":"Department of Applied Statistics, Institute of Mathematics and Basic Science, Hungarian University of Agriculture and Life Sciences (MATE), 1118 Budapest, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2404-0192","authenticated-orcid":false,"given":"Jerica","family":"Saboti\u010d","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Jo\u017eef Stefan Institute, 1000 Ljubljana, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,1,15]]},"reference":[{"key":"ref_1","first-page":"68","article-title":"Plant protease inhibitors: A defense strategy in plants","volume":"2","author":"Habib","year":"2007","journal-title":"Biotechnol. 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