Nep1-like proteins as a target for plant pathogen control

• DOI: 10.1371/journal.ppat.1009477
• Publication Date: 2021-04-16T11:32:52Z
• Authors: Katja Pirc; Vesna Hodnik; Tina Snoj; Tea Lenarčič; Simon Caserman; Marjetka Podobnik; Hannah Böhm; Isabell Albert; Anita Kotar; Janez Plavec; Jure Borišek; Martina Damuzzo; Alessandra Magist...; Boris Brus; Izidor Sosič; Stanislav Gobec; Thorsten Nürnberger; Gregor Anderluh
• Keywords: Phytophthora; Nicotiana; 2. Zero hunger; 0303 health sciences; QH301-705.5; NLP, pathogens, plants infections, inhibitors; Pythium; RC581-607; Molecular Dynamics Simulation; Surface Plasmon Resonance; Plant Leaves; Necrosis; 03 medical and health sciences; Tobacco; Immunologic diseases. Allergy; Biology (General); Research Article; Plant Diseases; Solanum tuberosum

The lack of efficient methods to control the major diseases of crops most important to agriculture leads to huge economic losses and seriously threatens global food security. Many of the most important microbial plant pathogens, including bacteria, fungi, and oomycetes, secrete necrosis- and ethylene-inducing peptide 1 (Nep1)-like proteins (NLPs), which critically contribute to the virulence and spread of the disease. NLPs are cytotoxic to eudicot plants, as they disturb the plant plasma membrane by binding to specific plant membrane sphingolipid receptors. Their pivotal role in plant infection and broad taxonomic distribution makes NLPs a promising target for the development of novel phytopharmaceutical compounds. To identify compounds that bind to NLPs from the oomycetesPythium aphanidermatumandPhytophthora parasitica, a library of 587 small molecules, most of which are commercially unavailable, was screened by surface plasmon resonance. Importantly, compounds that exhibited the highest affinity to NLPs were also found to inhibit NLP-mediated necrosis in tobacco leaves andPhytophthora infestansgrowth on potato leaves. Saturation transfer difference-nuclear magnetic resonance and molecular modelling of the most promising compound, anthranilic acid derivative, confirmed stable binding to the NLP protein, which resulted in decreased necrotic activity and reduced ion leakage from tobacco leaves. We, therefore, confirmed that NLPs are an appealing target for the development of novel phytopharmaceutical agents and strategies, which aim to directly interfere with the function of these major microbial virulence factors. The compounds identified in this study represent lead structures for further optimization and antimicrobial product development.