Poster # 307
John E. McLaughlin 1 and Nilgun E. Tumer 1
1. Rutgers University, Department of Plant Biology, New Brunswick, NJ
Corresponding Author: John McLaughlin, mclaughj@sebs.rutgers.edu
McLaughlin, John
Non-specific lipid transfer proteins (nsLTPs) are a diverse superfamily of small, cysteine-rich proteins that play key roles in plant defense, including direct inhibition of fungal pathogens such as Fusarium graminearum. Beyond their roles in cuticle formation and stress signaling, many nsLTPs act as antimicrobial peptides (AMPs), disrupting fungal membranes and limiting infection. Despite their importance, tens of thousands of nsLTP sequences in public databases remain uncharacterized, and the full extent of their structural diversity, particularly multi-domain variants, has not been systematically explored. We developed a robust bioinformatic workflow to identify, classify, and structurally model nsLTPs from large-scale sequence datasets. Starting with 12,880 proteins from the InterPro database (IPR000528), a custom Python script parsed the non-redundant dataset to detect all proteins containing one or more eight-cysteine motifs (8CMs). The analysis revealed 11,861 single-domain nsLTPs, 35 with two domains, three with three domains, and one protein with four domains. Among these, two multi-domain nsLTPs from African wild rice (Oryza barthii), A0A0D3HHF7 (two 8CMs) and A0A0D3HQM8 (four 8CMs), represent previously undescribed architectures within the family. Three-dimensional structures predicted using AlphaFold3 revealed compact yet highly variable multi-domain configurations, expanding the known structural landscape of nsLTPs. Because structure underpins antimicrobial function, these newly identified architectures may confer novel or enhanced activity against Fusarium and other fungal pathogens. This computational pipeline thus provides a foundation for discovering and prioritizing nsLTPs with superior antifungal potential for functional validation and, ultimately, deployment in breeding programs to reduce Fusarium head blight in wheat and barley.
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