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Payne, N.

Publications and source records attributed to Payne, N..

9 recordsLinked to original sources

A Gnotobiotic Rice Pathosystem Reveals How the Microbiome Suppresses Rice Blast Disease

Rice blast disease caused by the hemibiotrophic fungal pathogen Magnaporthe oryzae poses a major threat to global rice production, creating an urgent need for sustainable disease management strategies. Because plant-associated microbiomes can contribute to pathogen suppression and host defense, their role in rice blast resistance is of considerable interest. However, interactions among M. oryzae, rice, and the rice microbiome remain poorly understood. Using a gnotobiotic rice pathosystem, we investigated how microbiome colonization influences host responses to infection and how bacterial community structure changes following pathogen challenge. Microbiome colonization significantly reduced blast disease severity, while differential abundance analyses identified infection-associated shifts in bacterial community composition, including enrichment of Brevibacillus-associated taxa. Functional characterization of cultured endophytes revealed plant growth-promoting traits and antagonistic activity against M. oryzae. Two synthetic communities assembled from healthy- and infected-derived isolates significantly reduced disease severity, demonstrating that disease suppression can be recapitulated using simplified microbial communities. Interestingly, axenically grown plants exhibited elevated basal expression of multiple defense-associated genes but greater disease severity and reduced chitin-induced ROS production compared with holoxenic plants. In contrast, microbiome-colonized plants displayed enhanced ROS responsiveness and reduced disease, suggesting that the rice microbiome promotes a more effective and regulated immune state rather than simply increasing defense activation. Together, these findings advance our understanding of rice-microbiome-M. oryzae interactions and establish a tractable experimental platform for mechanistic studies of microbiome-mediated disease suppression.

microbiology↗

Biochemical and Mechanistic Characterization of the DNA-dependent Poly(ADP-ribose) Polymerase, MoPARP1, in Magnaporthe oryzae

Poly(ADP-ribose) polymerases (PARPs) are important regulators of DNA repair and cellular stress responses in eukaryotes. Although mammalian PARPs have been extensively characterized, comparatively little is known about PARPs in pathogenic filamentous fungi. Here, we define biochemical features of MoPARP1 from the plant-pathogenic fungus Magnaporthe oryzae. Mutation of the conserved catalytic glutamate E714 abolished detectable PARylation activity, whereas PAR generated by MoPARP1 was hydrolyzed by human PARG, suggesting synthesis of polymeric ADP-ribose. Several mammalian PARP inhibitors were effective against MoPARP1 in vitro, consistent with conservation of the catalytic inhibitor-binding pocket. DNA substrates containing 5' phosphate enhanced MoPARP1 catalytic activity, while both wild-type MoPARP1 and the catalytically inactive MoPARP1 E714A bound diverse DNA substrates. Quantitative analyses revealed that structurally distinct DNA substrates exhibit similar apparent binding affinities yet yield markedly different catalytic outputs, suggesting that DNA engagement alone does not determine productive PAR synthesis. MoPARP1 lacks the canonical N-terminal zinc-finger domains of human PARP1 and instead depends on its WGR-containing region for DNA association. Domain truncation analyses showed the BRCT-WGR region supports high-affinity DNA binding, whereas the WGR-PARP region retains catalytic competence despite weaker DNA affinity. Together, these findings establish a mechanistic framework for DNA-dependent PARylation in a filamentous fungal plant pathogen.

biochemistry↗

Pseudomonas effector AvrC is a rhamnosyltransferase with broad substrate specificity

Plant cells can directly or indirectly detect bacterial effectors, triggering a hypersensitive response to defend against pathogen infection. One extensively studied effector, AvrB, is a Fido (Fic, Doc, AvrB) domain-containing protein that acts as a glycosyltransferase. AvrC is an elusive Pseudomonas syringae avirulence effector protein with significant sequence and structural similarity to AvrB. Combining biochemistry, mass spectrometry, and AlphaFold prediction tools, we show that AvrC is a glycosyltransferase with auto-rhamnosylation activity. Like AvrB, AvrC can rhamnosylate a threonine residue (T166) on the A. thaliana guardee protein RIN4. In vitro assays revealed rhamnosylation substrates for AvrC also include plant coatomer subunits COPE1 and COPZ1. Collectively, our findings indicate that AvrC is a rhamnosyltransferase with broad substrate specificity. Our experimental strategies and findings provide valuable insights into future studies on the characterization of other Fido proteins.

biochemistry↗

Soil-derived Bacillus pumilus strains demonstrate antagonistic activity against Magnaporthe oryzae and multiple plant growth-promoting traits

Biological control strategies are increasingly being explored as sustainable alternatives for managing rice blast disease caused by Magnaporthe oryzae. In this study, we characterized three Bacillus pumilus isolates (DC01, DC09, and DC13) and evaluated their antifungal and plant-beneficial properties against M. oryzae. Whole genome sequencing revealed multiple biosynthetic gene clusters associated with the production of antimicrobial metabolites. All three isolates inhibited fungal growth in dual-culture assays, whereas heat-stable diffusible antifungal activity was primarily associated with the cell-free supernatants of DC09 and DC13. Exposure to bacterial supernatants disrupted fungal development, inducing abnormal hyphal morphology characterized by bulbous swelling, altered polarity, and increased branching in M. oryzae. Volatile organic compound assays further revealed that the DC isolates suppress fungal growth in the absence of physical contact. The isolates additionally inhibited the growth of other phytopathogenic fungi and selected human bacterial pathogens. All strains exhibited plant growth-promoting traits, including indole-3-acetic acid production and osmotic stress tolerance, whereas DC09 also displayed phosphate-solubilizing activity. Importantly, root inoculation with the DC isolates significantly reduced rice blast disease severity and induced expression of defense-associated genes involved in jasmonic acid/ethylene signaling and immune priming. Collectively, these findings identify the DC isolates, particularly DC09 and DC13, as promising multi-mechanistic biological control agents for sustainable rice blast management.

microbiology↗

Leveraging Chlorination-Based Mechanism for Resolving Subcellular Hypochlorous Acid

Hypochlorous acid (HOCl) is crucial for pathogen defense, but an imbalance in HOCl levels can lead to tissue damage and inflammation. Existing HOCl indicators employ an oxidation approach, which may not truly reveal the chlorinative stress environment. We designed a suite of indicators with a new chlorination-based mechanism, termed HOClSense dyes, to resolve HOCl in sub-cellular compartments. HOClSense dyes allow the visualization of HOCl with both switch-on and switch-off detection modes with diverse emission colors, as well as a unique redshift in emission. HOClSense features a minimalistic design with impressive sensing performance in terms of HOCl selectivity, and our design also facilitates functionalization through click chemistry for resolving subcellular HOCl. As a proof of concept, we targeted plasma membrane and lysosomes with HOClSense for subcellular HOCl mapping. With utilizing HOClSense, we discovered the STING pathway-induced HOCl production and the abnormal HOCl production in Niemann-Pick diseases. To the best of our knowledge, this is the first chlorination-based HOCl indicator series for resolving subcellular HOCl.

cell biology↗

Interface-guided phenotyping of coding variants in the transcription factor RUNX1 with SEUSS

Understanding the consequences of single amino acid substitutions in cancer driver genes remains an unmet need. Perturb-seq provides a tool to investigate the effects of individual mutations on cellular programs. Here we deploy SEUSS, a Perturb-seq like approach, to generate and assay mutations at physical interfaces of the RUNX1 Runt domain. We measured the impact of 115 mutations on RNA profiles in single myelogenous leukemia cells and used the profiles to categorize mutations into three functionally distinct groups: wild-type (WT)-like, loss-of-function (LOF)-like and hypomorphic. Notably, the largest concentration of functional mutations (non-WT-like) clustered at the DNA binding site and contained many of the more frequently observed mutations in human cancers. Hypomorphic variants shared characteristics with loss of function variants but had gene expression profiles indicative of response to neural growth factor and cytokine recruitment of neutrophils. Additionally, DNA accessibility changes upon perturbations were enriched for RUNX1 binding motifs, particularly near differentially expressed genes. Overall, our work demonstrates the potential of targeting protein interaction interfaces to better define the landscape of prospective phenotypes reachable by amino acid substitutions.

bioinformatics↗

Identifying the limits where variation in consumer stable isotope values reflect variation in diet

O_LIAnalysis of stable isotopes in cosumbers is used commonly to study their ecological and/or environmental niche. There is, however, considerable debate regarding how isotopic values relate to diet and how other sources of variation confound this link, which can undermine the utility. C_LIO_LIFrom the analysis of a simple, but general, model of isotopic incorporation in consumer organisms, we examine the relationship between isotopic variance among individuals, and diet variability within a consumer population. C_LIO_LIWe show that variance in consumer isotope values is directly proportional to variation in diet (through Simpson indices), to the number of isotopically distinct food sources in the diet, and to the baseline variation within and among the isotope values of the food sources. Additionally, when considering temporal diet variation within a consumer we identify the interplay between diet turnover rates and tissue turnover rates that controls the sensitivity of stable isotopes to detect diet variation. C_LIO_LIOur work demonstrates that variation in the stable isotope values of consumers reflect variation in their diet. This relationship, however, can be confounded with other factors to the extent that they may mask the signal coming from diet. We show how simple quantitative corrections can recover a direct 1:1 correlation in some situations, and in others we can adjust our interpretation in light of the new understanding arising from our models. Our framework provides guidance for the design and analysis of empirical studies where the goal is to infer niche width from stable isotope data. C_LI

ecology↗

Body size as a magic trait in two plant-feeding insect species

When gene flow accompanies speciation, recombination can decouple divergently selected loci and loci conferring reproductive isolation. This barrier to sympatric divergence disappears when assortative mating and disruptive selection involve the same "magic" trait. Although magic traits could be widespread, the relative importance of different types of magic traits to speciation remains unclear. Because body size frequently contributes to host adaptation and assortative mating in plant-feeding insects, we evaluated several magic trait predictions for this trait in a pair of sympatric Neodiprion sawfly species adapted to different pine hosts. A large morphological dataset revealed that sawfly adults from populations and species that use thicker-needled pines are consistently larger than those that use thinner needled-pines. Fitness data from recombinant backcross females revealed that egg size is under divergent selection between the preferred pines. Lastly, mating assays revealed strong size-assortative mating within and between species in three different crosses, with the strongest prezygotic isolation between populations that have the greatest interspecific size differences. Together, our data support body size as a magic trait in pine sawflies and possibly many other plant-feeding insects. Our work also demonstrates how intraspecific variation in morphology and ecology can cause geographic variation in the strength of prezygotic isolation.

evolutionary biology↗

A miniaturized threshold-triggered acceleration data-logger for recording burst movements of aquatic animals

Animal-borne accelerometers are effective tools for quantifying the kinematics of animal behaviors, such as swimming, running, and flying, under natural conditions. However, quantifying burst movements of small and agile aquatic animals (e.g., small teleost fish), such as during predatory behavior, or while fleeing, remains challenging. To capture the details of burst movements, accelerometers need to sample at a very high frequency, which will inevitably shorten the duration of the recording or increase the size of the device. To overcome this problem, we developed a high-frequency acceleration data-logger that can be triggered by a manually-defined acceleration threshold, thus allowing the selective measurement of animal burst movements. We conducted experiments under laboratory and field conditions to examine the performance of the logger. The laboratory experiment using red seabream (Pagrus major) showed that the new logger could measure the kinematics of their escape behaviors (i.e., body beat cycles and maximum acceleration values). The field experiment using free-swimming yellowtail kingfish (Seriola lalandi) showed that the loggers trigger correctly (i.e., of the 18 burst movements, 17 were recorded by the loggers). We suggest that this new logger can be applied to measure the burst movements of various small and agile animals, whose movements may be otherwise difficult to measure.

animal behavior and cognition↗