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Carabrone, a botanical bicyclic sesquiterpenic lactone, is a promising fungicidal agent that can effectively control <jats:italic>G.\u00a0tritici<\/jats:italic>. However, the mechanism of action of carabrone against <jats:italic>G.\u00a0tritici<\/jats:italic> remains largely unclear. Here, we used immunogold for subcellular localization of carabrone and the results showed that carabrone is subcellularly localized in the mitochondria of <jats:italic>G.\u00a0tritici<\/jats:italic>. We then explored the functional analysis of genes <jats:italic>GtCytc<\/jats:italic><jats:sub>1<\/jats:sub>, <jats:italic>GtCytb<\/jats:italic>, and <jats:italic>GtIsp<\/jats:italic> of the mitochondrial respiratory chain cytochrome <jats:italic>bc<\/jats:italic><jats:sub>1<\/jats:sub> complex in <jats:italic>G.\u00a0tritici<\/jats:italic> by RNA silencing as a possible target of carabrone. The results showed that the silenced mutant \u2206GtIsp is less sensitive to carabrone compared to \u2206GtCytc<jats:sub>1<\/jats:sub> and \u2206GtCytb. Compared with the control, the activities of complex III in all the strains, except \u2206GtIsp and carabrone\u2010resistant isolate 24\u2010HN\u20101, were significantly decreased following treatment with carabrone at EC<jats:sub>20<\/jats:sub> and EC<jats:sub>80<\/jats:sub> in vitro (40%\u201350% and 70%\u201380%, respectively). The activities of mitochondrial respiratory chain complex III and the mitochondrial respiration oxygen consumption rates in all the strains, except \u2206GtIsp and 24\u2010HN\u20101, were higher with respect to the control when treated with carabrone at EC<jats:sub>20<\/jats:sub> in vivo. The rates of mitochondrial respiration of all strains, except \u2206GtIsp, were significantly inhibited following treatment with carabrone at EC<jats:sub>80<\/jats:sub> (ranging from 57% to 81%). This study reveals that the targeting of the iron\u2013sulphur protein encoded by <jats:italic>GtIsp<\/jats:italic> is highly sensitive to carabrone and provides a direction for the research of carabrone's target.<\/jats:p>","DOI":"10.1111\/mpp.12993","type":"journal-article","created":{"date-parts":[[2020,9,30]],"date-time":"2020-09-30T12:51:11Z","timestamp":1601470271000},"page":"1529-1544","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["A functional analysis of mitochondrial respiratory chain cytochrome <i>bc<\/i><sub>1<\/sub> complex in <i>Gaeumannomyces tritici<\/i> by RNA silencing as a possible target of carabrone"],"prefix":"10.1111","volume":"21","author":[{"given":"Mei","family":"Wang","sequence":"first","affiliation":[{"name":"Engineering and Research Center of Biological Pesticide of Shaanxi Province Northwest A&amp;F University Yangling China"},{"name":"College of Life Sciences Yulin University Yulin 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