Abstract
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 oomycetes Pythium aphanidermatum and Phytophthora 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 and Phytophthora infestans growth 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.
Highlights
Plant pathogens cause diverse diseases that affect crop yield and food quality, which leads to extensive annual financial losses worldwide
necrosis- and ethylene-inducing peptide 1 (Nep1)-like proteins (NLPs) constitute a large protein family of virulent agents that are prevalent in different microbial taxa such as bacteria, oomycetes, and fungi
We have identified three molecules that bind to NLPs in the micromolar range and reduce NLPinduced necrosis in tobacco leaves
Summary
Plant pathogens cause diverse diseases that affect crop yield and food quality, which leads to extensive annual financial losses worldwide. Approximately 50% of wheat and more than 80% of cotton production would be eliminated by different diseases in the absence of agrochemical interventions [1]. Controlling plant pathogens is of crucial importance for modern agriculture. Fungicides are widely used in developed agricultural production to control diseases and maintain sufficient crop yield and product quality. The mechanism of action is not known for most fungicides, and there may be possible side effects for the host plants. The use of nonspecific chemical strategies to control crop production can have deleterious effects on the environment and human health [2]. New compounds and strategies with better effectiveness, lower application dosage, higher selectivity, and fewer costs and lower environmental impact are highly desirable
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