Recombinant expression, purification, and antifungal activity of the novel antimicrobial peptide TaW662

Abstract

AbstractBlumeria graminisf. sp.tritici(Bgt) is a significant wheat fungal pathogen, posing threats to both yield and quality. Antimicrobial peptides, with their broad-spectrum activities, hold promise in combatingBgt-induced wheat fungal diseases.In this study, we identifiedTaW662, an antifungal peptide gene sourced from the wheat-Thinopyrum intermediumdisomic alien addition line SN6306. Through third-generation transcriptome sequencing, we obtained the full-length transcript ofTaW662. Notably,TaW662exhibited upregulated expression in response to powdery mildew infection in SN6306. Subcellular localization analysis revealed TaW662’s extracellular secretion, suggesting its role in defense mechanisms. Additionally, the TaW662 protein was expressed inEscherichia coli, and the purified protein could inhibit the growth ofBgt in vitro. Utilizing the online alphafold2 server, we predicted the three-dimensional structure of TaW662, aiding in understanding its fungicidal mechanisms. Analysis of TaW662’s physicochemical properties further supported its potential efficacy as a fungicide againstBgt. In conclusion, TaW662 emerges as a promising candidate for combatingBgt-induced wheat fungal diseases, warranting further exploration for agricultural disease management strategies.HighlightsTaW662, a secreted protein homologous toTaWIR1, is induced in wheat byBlumeria graminisf. sp.tritici(Bgt).The expression pattern ofTaW662in wheat under induced byBgtwas analysed using RNA-Seq technology.The three-dimensional structure of TaW662 was predicted using AlphaFold2.The growth ofBgtis inhibited by recombinant TaW662.