DOI: 10.6084/m9.figshare.31430149
title: <b>Integrated omics analyses reveal natamycin disrupts the amino acid and oxidative phosphorylation pathways in </b><b><i>Magnaporthe oryzae</i></b>

description: This research paper, titled <b>"Integrated omics analyses reveal natamycin disrupts the amino acid and oxidative phosphorylation pathways in</b><b> </b><b><i>Magnaporthe oryzae</i></b><b>,"</b> investigates the antifungal potential and mechanism of action of natamycin against the rice blast fungus.<b>The Problem:</b> Rice blast, caused by the fungus <i>Magnaporthe oryzae</i>, is a devastating disease threatening global rice production. Current control methods rely heavily on chemical pesticides, which have environmental and safety drawbacks, and on resistant crop varieties, which the pathogen can eventually overcome.<b>The Compound:</b> Natamycin is a natural antimicrobial compound produced by <i>Streptomyces</i> bacteria, widely used as a food preservative. However, its specific effects on <i>M. oryzae</i> were not well understood.<b>Objective:</b> The study aimed to evaluate natamycin's efficacy against <i>M. oryzae</i> and, more importantly, to uncover its precise mode of action using a combination of transcriptomic (gene expression) and metabolomic (metabolite profiling) analyses.Key Findings & Results<b>A. Strong Antifungal Activity</b>Natamycin effectively inhibited the growth of <i>M. oryzae</i> in the lab, with an EC₅₀ (half-maximal effective concentration) of <b>11.77 mg/L</b>.In greenhouse experiments on rice seedlings, natamycin treatment drastically reduced the severity of rice blast disease, lowering the disease index from 64.43% in untreated plants to just <b>4.1%</b> in treated plants.<b>B. Mechanism of Action Revealed by Multi-Omics</b><br>The core of the study reveals that natamycin works through a multi-target mechanism, primarily by disrupting two critical cellular processes:<b>Disruption of Oxidative Phosphorylation (Energy Production):</b><b>Transcriptomics:</b> Genes encoding key components of the oxidative phosphorylation pathway (the cell's energy-producing system in mitochondria) were significantly <b>downregulated</b>. This includes genes for ATP synthase, cytochrome c oxidase, and NADH dehydrogenase.<b>Metabolomics:</b> The levels of adenosine diphosphate (ADP), the molecule used to make ATP, were significantly reduced, confirming a blockage in energy production.<b>Functional Validation:</b> When the fungus was genetically engineered to overexpress two specific oxidative phosphorylation genes, <i>MGG_17901</i> (cytochrome c oxidase subunit) and <i>MGG_04969</i> (electron transfer protein), it became significantly more <b>tolerant</b> to natamycin. This proves these genes are key targets of the drug. Overexpressing these genes also made the fungus more virulent on rice plants.<b>Mechanistic Link:</b> Natamycin treatment was shown to disrupt the balance of reactive oxygen species (ROS), leading to oxidative stress, which is a direct consequence of a malfunctioning electron transport chain.<b>Disruption of Amino Acid Biosynthesis (Building Blocks for Growth):</b>Multiple amino acid biosynthesis pathways (e.g., valine, leucine, isoleucine, arginine, proline) were suppressed.This was evident at both the gene expression level (e.g., downregulation of enzymes like threonine dehydratase, acetolactate synthase) and the metabolite level (e.g., depletion of amino acids like proline and intermediates like spermidine).This disruption starves the fungus of essential building blocks for protein synthesis and growth.<b>Molecular Basis of Interaction</b><b>Molecular docking</b> simulations predicted that natamycin can physically bind to the proteins encoded by the target genes <i>MGG_17901</i> and <i>MGG_04969</i> via hydrogen bonds.Sequence analysis of 96 different <i>M. oryzae</i> strains showed that the binding sites for natamycin on these proteins are <b>highly conserved</b>, suggesting that resistance to this mechanism is unlikely to develop easily.