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Specialist herbivores that feed on brassicaceous plants have evolved various mechanisms aimed at preventing the formation of toxic isothiocyanates. In contrast, generalist herbivores typically detoxify isothiocyanates through glutathione conjugation upon exposure. Here, we examined the response of an extreme generalist herbivore, the two-spotted spider mite Tetranychus urticae (Koch), to indole glucosinolates. Tetranychus urticae is a composite generalist whose individual populations have a restricted host range but have an ability to rapidly adapt to initially unfavorable plant hosts. Through comparative transcriptomic analysis of mite populations that have differential susceptibilities to Arabidopsis defenses, we identified \u03b2-cyanoalanine synthase of T. urticae (TuCAS), which encodes an enzyme with dual cysteine and \u03b2-cyanoalanine synthase activities. We combined Arabidopsis genetics, chemical complementation and mite reverse genetics to show that TuCAS is required for mite adaptation to Arabidopsis through its \u03b2-cyanoalanine synthase activity. Consistent with the \u03b2-cyanoalanine synthase role in detoxification of hydrogen cyanide (HCN), we discovered that upon mite herbivory, Arabidopsis plants release HCN. We further demonstrated that indole glucosinolates are sufficient for cyanide formation. Overall, our study uncovered Arabidopsis defenses that rely on indole glucosinolate-dependent cyanide for protection against mite herbivory. In response, Arabidopsis-adapted mites utilize the \u03b2-cyanoalanine synthase activity of TuCAS to counter cyanide toxicity, highlighting the mite\u2019s ability to activate resistant traits that enable this extreme polyphagous herbivore to exploit cyanogenic host plants.<\/jats:p>","DOI":"10.1093\/plphys\/kiac147","type":"journal-article","created":{"date-parts":[[2022,3,25]],"date-time":"2022-03-25T12:22:58Z","timestamp":1648210978000},"page":"1961-1975","source":"Crossref","is-referenced-by-count":20,"title":["\u03b2-Cyanoalanine synthase protects mites against Arabidopsis defenses"],"prefix":"10.1093","volume":"189","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4492-8607","authenticated-orcid":false,"given":"Sameer","family":"Dixit","sequence":"first","affiliation":[{"name":"Department of Biology, The University of Western Ontario , London, Ontario, Canada N6A 5B7"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6732-5497","authenticated-orcid":false,"given":"Emilie","family":"Widemann","sequence":"additional","affiliation":[{"name":"Department of Biology, The University of Western Ontario , London, Ontario, Canada N6A 5B7"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicolas","family":"Bensoussan","sequence":"additional","affiliation":[{"name":"Department of Biology, The University of Western Ontario , London, Ontario, Canada N6A 5B7"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3383-834X","authenticated-orcid":false,"given":"Golnaz","family":"Salehipourshirazi","sequence":"additional","affiliation":[{"name":"Department of Biology, The University of Western Ontario , London, Ontario, Canada N6A 5B7"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7913-7920","authenticated-orcid":false,"given":"Kristie","family":"Bruinsma","sequence":"additional","affiliation":[{"name":"Department of Biology, The University of Western Ontario , London, Ontario, Canada N6A 5B7"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8248-8884","authenticated-orcid":false,"given":"Maja","family":"Milojevic","sequence":"additional","affiliation":[{"name":"Department of Biology, The University of Western Ontario , London, Ontario, Canada N6A 5B7"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7155-7381","authenticated-orcid":false,"given":"Akanchha","family":"Shukla","sequence":"additional","affiliation":[{"name":"Department of Biology, The University of Western Ontario , London, Ontario, Canada N6A 5B7"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2414-4813","authenticated-orcid":false,"given":"Luis C","family":"Romero","sequence":"additional","affiliation":[{"name":"Instituto de Bioqu\u00edmica Vegetal y Fotos\u00edntesis, Consejo Superior de Investigaciones Cient\u00edficas and Universidad de Sevilla , E-41092 Seville, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7120-4655","authenticated-orcid":false,"given":"Vladimir","family":"Zhurov","sequence":"additional","affiliation":[{"name":"Department of Biology, The University of Western Ontario , London, Ontario, Canada N6A 5B7"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1209-3200","authenticated-orcid":false,"given":"Mark A","family":"Bernards","sequence":"additional","affiliation":[{"name":"Department of Biology, The University of Western Ontario , London, Ontario, Canada N6A 5B7"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7521-5485","authenticated-orcid":false,"given":"Maksymilian","family":"Chruszcz","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry, University of South Carolina , Columbia, South Carolina, 29208, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4214-2336","authenticated-orcid":false,"given":"Miodrag","family":"Grbi\u0107","sequence":"additional","affiliation":[{"name":"Department of Biology, The University of Western Ontario , London, Ontario, Canada N6A 5B7"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6570-8796","authenticated-orcid":false,"given":"Vojislava","family":"Grbi\u0107","sequence":"additional","affiliation":[{"name":"Department of Biology, The University of Western Ontario , London, Ontario, Canada N6A 5B7"},{"name":"Instituto de 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