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To obtain effective disease resistance genes against cotton Verticillium wilt, we constructed several fusion genes, in which binding domains targeting chitin, sphingolipid or ergosterol in the fungal cell wall or cell membrane were individually fused to the antifungal peptide BbAFP1 from entomopathogenic fungus <jats:italic>Beauveria bassiana<\/jats:italic>. Transient expression of fusion genes in cotton cotyledons indicated that the BbAFP1::ErBD fusion peptide with an ergosterol binding domain exhibited better disease resistance against <jats:italic>V.\u00a0dahliae<\/jats:italic> than wild\u2010type <jats:italic>BbAFP1<\/jats:italic> and other fusion genes. <jats:italic>BbAFP1::ErBD<\/jats:italic> and <jats:italic>BbAFP1<\/jats:italic> transgenic cotton were obtained and verified by Southern and Western blotting. Compared with BbAFP1\u2010expressing cotton, BbAFP1::ErBD\u2010expressing cotton showed higher disease resistance against <jats:italic>V.\u00a0dahliae<\/jats:italic>, with smaller lesion areas (0.07\u00a0cm<jats:sup>2<\/jats:sup> vs. 0.16\u00a0cm<jats:sup>2<\/jats:sup>) on the leaves and a lower disease index (23.9 vs. 34.5). Overexpression of <jats:italic>BbAFP1::ErBD<\/jats:italic> by transgenic tobacco also showed enhanced disease resistance against <jats:italic>V.\u00a0dahliae<\/jats:italic> compared with that of the wild\u2010type gene. These results indicated that construction of fusion antifungal peptides that target fungal cells is a powerful strategy to obtain new anti\u2010disease genes, and the obtained fusion gene <jats:italic>BbAFP1::ErBD<\/jats:italic> has the potential to defend against plant fungal diseases.<\/jats:p>","DOI":"10.1111\/pbi.13517","type":"journal-article","created":{"date-parts":[[2020,11,21]],"date-time":"2020-11-21T06:51:11Z","timestamp":1605941471000},"page":"926-936","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Ergosterol\u2010targeting fusion antifungal peptide significantly increases the Verticillium wilt resistance of cotton"],"prefix":"10.1111","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1178-3998","authenticated-orcid":false,"given":"Sheng","family":"Tong","sequence":"first","affiliation":[{"name":"Biotechnology Research Center Chongqing Key Laboratory of Plant Resource Conservation and 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