Recombinant Pseudomonas syringae pv. tomato Urease accessory protein UreE 1 (ureE1)

What is the role of UreE1 in Pseudomonas syringae pv. tomato?

UreE1 functions as an accessory protein that assists in the assembly of the urease enzyme complex in Pseudomonas syringae pv. tomato. As a nickel-binding chaperone, it facilitates the incorporation of nickel ions into the urease active site, which is essential for the enzyme's catalytic activity. In the context of plant pathogenicity, urease activity enables the bacterium to utilize urea as a nitrogen source during colonization of the host plant's apoplast, potentially contributing to its virulence by modifying the local pH and creating favorable conditions for bacterial growth . The protein is part of the broader metabolic machinery that allows P. syringae pv. tomato to establish infection and cause bacterial speck disease in tomato plants.

How does UreE1 contribute to P. syringae pv. tomato pathogenicity?

While UreE1 is not a direct virulence factor like the Type III Secretion System (T3SS) effectors such as HopM1, AvrE1, AvrPto1, and AvrPtoB that are critical for P. syringae pv. tomato pathogenicity , it plays a supportive role in bacterial fitness during infection. By ensuring functional urease activity, UreE1 indirectly contributes to:

• Nitrogen metabolism during the apoplastic colonization phase

• Potential alkalization of the infection site through ammonia production from urea hydrolysis

• Enhanced bacterial survival under acidic stress conditions within the plant tissue

• Possible modulation of host defense responses through altered pH conditions

This metabolic functionality complements the direct virulence mechanisms that P. syringae pv. tomato employs to suppress pattern-triggered immunity (PTI) and establish successful infections in tomato plants .

What methods are commonly used to purify recombinant UreE1 from P. syringae pv. tomato?

Recombinant UreE1 purification typically follows these methodological steps:

• Cloning: The ureE1 gene is amplified from P. syringae pv. tomato genomic DNA using PCR with specific primers designed based on the published sequence.

• Expression vector construction: The gene is inserted into an appropriate expression vector (e.g., pET system) containing a histidine tag for purification.

• Transformation: The construct is transformed into an E. coli expression strain such as BL21(DE3).

• Protein expression: IPTG induction at optimized temperature (typically 16-28°C) to minimize inclusion body formation.

• Cell lysis: Bacterial cells are disrupted by sonication or French press in a buffer containing protease inhibitors.

• Affinity chromatography: His-tagged UreE1 is purified using Ni-NTA or IMAC columns.

• Size exclusion chromatography: Further purification to remove aggregates and ensure homogeneity.

• Quality assessment: SDS-PAGE and Western blot analysis to confirm purity and identity.

For functional studies, it's essential to ensure the recombinant protein retains nickel-binding ability, which can be verified through isothermal titration calorimetry (ITC) or metal-binding assays using colorimetric reagents.

What recombineering approaches are effective for modifying the ureE1 gene in P. syringae pv. tomato?

The most effective recombineering approaches for modifying the ureE1 gene in P. syringae pv. tomato utilize the RecTE system, which has been specifically optimized for this bacterium. The RecT protein from P. syringae promotes recombination of single-stranded DNA oligonucleotides, while efficient recombination of double-stranded DNA requires the expression of both RecT and RecE homologs . This system enables targeted gene modifications including:

• Allelic replacement: For creating precise point mutations in ureE1

• Gene deletions: For functional studies of UreE1's role in urease activity

• Reporter fusions: For studying ureE1 expression patterns

• Epitope tagging: For localization and interaction studies

The methodology involves:

• Constructing a recombineering plasmid expressing RecTE from P. syringae (like pUCP24/RecTE)

• Designing homology arms (50-100 bp) flanking the target modification site

• Introducing linear DNA by electroporation into P. syringae cells expressing RecTE

• Selection of recombinants using appropriate markers

• Elimination of the RecTE expression vector using the sacB counterselection system

This approach has been shown to achieve recombination frequencies sufficient for direct selection of modified cells without the need for complex selection schemes.

How can researchers optimize heterologous expression of UreE1 to maintain its structural integrity?

Optimizing heterologous expression of UreE1 to maintain structural integrity requires addressing several critical factors:

Protein production should be monitored by SDS-PAGE analysis of soluble and insoluble fractions, with optimization of conditions to maximize the proportion in the soluble fraction. Once purified, circular dichroism (CD) spectroscopy can confirm proper secondary structure, while thermal shift assays can assess stability under different buffer conditions. For functional validation, nickel-binding assays and urease activation assays using the recombinant protein with urease apoenzyme should be performed to ensure the expressed protein retains its native chaperoning function.

What are the challenges in expressing UreE1 in plant systems for functional studies?

Expressing functional UreE1 in plant systems presents several significant challenges:

• Codon optimization requirements: The GC-rich bacterial codons need adjustment for efficient expression in plant systems.

• Subcellular localization issues: Bacterial UreE1 lacks plant-specific targeting sequences, necessitating the addition of appropriate transit peptides for targeting to relevant compartments.

• Potential cytotoxicity: Disruption of metal homeostasis in plant cells may occur due to the nickel-binding properties of UreE1.

• Host defense responses: Plants may recognize bacterial proteins and trigger immunity responses that could interfere with experimental outcomes, particularly as P. syringae is a known plant pathogen .

• Functional partners absence: UreE1 functions in a complex with other urease assembly proteins that may not be present in plants.

To address these challenges, researchers can:

• Use inducible expression systems to control protein accumulation levels

• Employ tissue-specific promoters to restrict expression to relevant tissues

• Co-express other components of the urease activation complex

• Consider fusion tags that enhance stability in planta

• Use confocal microscopy with fluorescent tags to monitor localization and potential aggregation

Additionally, assessing whether plant expression affects host defense pathways is crucial, as immune responses could confound functional studies by altering plant physiology independently of UreE1's direct effects.

What techniques are most reliable for analyzing UreE1 metal-binding properties?

The most reliable techniques for analyzing UreE1 metal-binding properties include a combination of biophysical and biochemical approaches:

• Isothermal Titration Calorimetry (ITC) provides direct measurement of binding thermodynamics, including:

  • Binding stoichiometry (number of Ni²⁺ ions bound per UreE1 dimer)

  • Binding affinity (Kd values)

  • Enthalpy (ΔH) and entropy (ΔS) changes

• Differential Scanning Calorimetry (DSC) assesses thermal stability changes upon metal binding, revealing how nickel coordination affects protein structure.

• Circular Dichroism (CD) Spectroscopy with and without nickel ions detects secondary structure changes induced by metal binding.

• X-ray Absorption Spectroscopy (XAS), specifically X-ray absorption near-edge structure (XANES) and extended X-ray absorption fine structure (EXAFS), provides detailed information about the coordination chemistry of the nickel-binding site.

• Equilibrium Dialysis coupled with inductively coupled plasma mass spectrometry (ICP-MS) for precise quantification of bound metal.

• Fluorescence Spectroscopy using intrinsic tryptophan fluorescence or fluorescent metal probes can track conformational changes associated with metal binding.

• Size Exclusion Chromatography with Multi-Angle Light Scattering (SEC-MALS) to assess whether metal binding affects oligomerization state.

Researchers should employ multiple complementary methods, as each provides different insights into the metal-binding mechanism. For example, ITC data on UreE1-nickel interactions should be validated with structural studies to confirm binding site architecture and coordination geometry.

How does UreE1 interact with other urease accessory proteins in the urease maturation pathway?

UreE1 functions within a complex network of protein-protein interactions in the urease maturation pathway. The interaction hierarchy follows this general pattern:

• Initial complex formation: UreE1 binds nickel ions through its histidine-rich C-terminal domain.

• UreE1-UreG interaction: UreE1 transfers nickel to the GTPase UreG, which acts as an energy-providing component.

• Ternary complex assembly: UreE1-UreG associates with UreF-UreH/UreD to form the UreE-UreF-UreG-UreH complex.

• Nickel transfer to urease: This complex ultimately delivers nickel to the urease apoprotein (UreABC).

These interactions can be characterized using techniques such as:

• Surface Plasmon Resonance (SPR) to determine binding kinetics

• Co-immunoprecipitation to confirm interactions in cell lysates

• Bacterial Two-Hybrid (B2H) assays for in vivo interaction verification

• Crosslinking studies coupled with mass spectrometry to identify interaction interfaces

• Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS) to map conformational changes during complex assembly

Mutations in the metal-binding domain of UreE1 disrupt the interaction cascade and prevent proper urease activation, highlighting the critical role of UreE1 in the nickel delivery pathway. Understanding these interaction networks is essential for developing potential inhibitors that could target bacterial urease activity as an antimicrobial strategy against P. syringae pv. tomato.

What computational approaches are most effective for predicting UreE1 structure-function relationships?

Computational approaches for predicting UreE1 structure-function relationships should integrate multiple modeling techniques and validation strategies:

• Homology Modeling/AlphaFold2 Prediction:

  • Based on structural homology with characterized UreE proteins from related bacteria

  • Validation through Ramachandran plot analysis, QMEAN, and ProSA-web scores

• Molecular Dynamics (MD) Simulations:

  • Assess conformational stability and flexibility

  • Identify metal-binding site dynamics in aqueous environments

  • Reveal allosteric effects of nickel binding on distant protein regions

• Quantum Mechanics/Molecular Mechanics (QM/MM):

  • Provide detailed characterization of electron distribution around the metal-binding site

  • Calculate energetics of nickel coordination

• Protein-Protein Docking:

  • Predict interaction interfaces with UreG, UreF, and other partners

  • Evaluate binding energy landscapes and identify critical residues

• Evolutionary Coupling Analysis:

  • Identify co-evolving residues suggesting functional interactions

  • Direct Coupling Analysis (DCA) can predict contacts within protein structure

• Machine Learning Integration:

  • Combine sequence features, predicted structures, and experimental data

  • Classify critical functional residues based on conservation patterns

These computational approaches should be integrated into a workflow that proceeds from sequence analysis to structural modeling, followed by dynamic simulations and interaction predictions. The results should guide experimental design for site-directed mutagenesis to validate computationally predicted functional residues.

How does UreE1 expression vary across different strains of P. syringae pv. tomato and during different infection stages?

UreE1 expression patterns show significant variation across P. syringae pv. tomato strains and infection stages, reflecting its role in bacterial adaptation to the host environment:

Expression analysis through qRT-PCR and RNA-seq studies reveals that UreE1 transcription is typically coordinated with other urease components but follows distinct patterns from the primary virulence factors like T3SS effectors . While the T3SS genes are highly expressed during the early stages of infection to suppress host immunity, UreE1 expression increases gradually as the bacteria establish in the apoplast where nitrogen metabolism becomes critical for sustained growth.

Factors affecting UreE1 expression include:

• pH changes in the infection microenvironment

• Nitrogen source availability in the apoplast

• Stress conditions encountered during colonization

• Plant defense responses that alter the infection site chemistry

The expression patterns also differ between strains with different virulence profiles. For example, highly aggressive strains tend to show earlier upregulation of UreE1, potentially reflecting more rapid adaptation to the host environment. These expression differences may contribute to the variable virulence observed between race 0 and race 1 strains of P. syringae pv. tomato in tomato plants carrying the Pto resistance gene .

What methods are most effective for studying UreE1 function during P. syringae infection of tomato plants?

The most effective methods for studying UreE1 function during P. syringae infection of tomato plants combine molecular genetics, imaging techniques, and physiological assays:

• Gene Knockout and Complementation Studies:

  • Generate ureE1 deletion mutants using RecTE-based recombineering

  • Complement with wild-type or mutant versions under native or inducible promoters

  • Compare infection progression and bacterial proliferation in planta

• Reporter Systems:

  • Construct transcriptional fusions (ureE1 promoter-GFP/LUX)

  • Create translational fusions (UreE1-fluorescent protein) that maintain functionality

  • Monitor expression timing and localization during infection

• In Planta Bacterial Transcriptomics:

  • RNA-seq from infected plant tissue with bacterial RNA enrichment

  • Compare ureE1 expression with other virulence and metabolism genes

  • Correlate expression with infection stages

• Metabolic Profiling:

  • Measure urease activity in planta during infection

  • Quantify urea and ammonia levels in infected tissues

  • Monitor pH changes in the apoplastic fluid

• Confocal Microscopy with pH Indicators:

  • Visualize pH changes around bacterial colonies in leaf tissue

  • Correlate with UreE1 expression and urease activity

• Plant Response Assessment:

  • Compare defense gene expression against wild-type and ureE1 mutants

  • Evaluate ion leakage and reactive oxygen species production

  • Assess callose deposition and other immunity markers

These methodologies should be applied in a coordinated experimental design that follows the infection timeline, from initial bacterial entry through symptom development. Comparing wild-type bacteria with ureE1 mutants across various tomato genotypes (susceptible and resistant) provides comprehensive insights into UreE1's contribution to pathogenesis.

How does environmental stress affect UreE1 function in P. syringae survival outside the host?

Environmental stress significantly modulates UreE1 function and contributes to P. syringae survival in non-host environments. The primary environmental factors affecting UreE1 function include:

• UV Radiation Exposure:
  P. syringae strains experience significant UV exposure in agricultural settings and during airborne dispersal. Studies show that UV-C exposure at 60 J/m² reduces bacterial populations to less than 1% of initial counts . UreE1 expression increases upon UV stress, potentially as part of a general stress response. The increased urease activity may help neutralize acid stress caused by UV-induced metabolic disruption.

• Desiccation Stress:
  Under low relative humidity conditions (RH <5% and 33%), P. syringae survival depends on metabolic adaptations . UreE1-mediated urease activity provides ammonia that acts as a compatible solute, helping to maintain cellular hydration. Additionally, ammonia production can neutralize acids that concentrate during cell dehydration.

• Temperature Fluctuations:
  During temperature cycling between freezing and thawing, which P. syringae commonly experiences in environmental reservoirs, urease activity provides metabolic flexibility. UreE1 function is particularly important during the recovery phase after freezing stress, when rapid nitrogen metabolism supports cellular repair.

• Nutrient Limitation:
  In soil and water environments with limited nitrogen, UreE1-activated urease allows P. syringae to utilize urea as an alternative nitrogen source, providing a competitive advantage over microorganisms lacking this metabolic capability.

The environmental fitness conferred by UreE1 may explain why urease genes are maintained in the core genome of P. syringae strains despite variability in other genetic elements like effector proteins and prophages that differ between race 0 and race 1 strains . This suggests UreE1's role extends beyond pathogenicity to general bacterial fitness in diverse environments.

How can researchers resolve contradictory findings about UreE1 function from different experimental approaches?

When faced with contradictory findings about UreE1 function from different experimental approaches, researchers should employ a systematic resolution strategy:

• Methodological Reconciliation Framework:

  Contradictions between qualitative and quantitative findings often stem from methodological differences rather than actual biological disparities . For UreE1 research, this frequently manifests when in vitro biochemical studies show different results than in vivo functional analyses.

• Critical Assessment of Assay Conditions:

  Approach	Potential Limitations	Verification Strategies
  In vitro nickel-binding assays	Non-physiological buffer conditions	Test under conditions mimicking apoplastic environment
  Heterologous expression studies	Missing bacterial cofactors	Co-express other urease accessory proteins
  Plant infection phenotyping	Compensatory mechanisms mask effects	Use multiple knockout combinations
  Structural predictions	Incomplete validation	Verify with experimental structure determination

• Integrative Experimental Design:

  Rather than viewing contradictory results as problematic, design experiments that intentionally employ both approaches to capture a more complete picture. For example, if biochemical assays suggest UreE1 binds two nickel ions but crystallography shows only one binding site, isothermal titration calorimetry combined with size exclusion chromatography could resolve whether dimerization affects binding stoichiometry.

• Contextual Factors Analysis:

  Different P. syringae strains may show varying UreE1 functions based on their genetic background. Comparative analysis across multiple strains, including both race 0 and race 1 variants , can help identify strain-specific factors that influence UreE1 behavior.

• Reproducibility Assessment:

  Ensure findings can be reproduced by independent researchers using identical methods. For contradictory results, collaborative cross-validation between research groups can identify subtle methodological differences affecting outcomes.

Remember that what appears contradictory may actually represent different facets of UreE1's complex biological role. As noted by research methodologists, "in any given case either [approach] may be correct or both may be correct—about somewhat different questions—even though the findings may seem contradictory" .

What are the most promising approaches for targeting UreE1 in antimicrobial development against P. syringae?

Targeting UreE1 for antimicrobial development against P. syringae represents a promising strategy that focuses on disrupting bacterial metabolism rather than directly attacking primary virulence mechanisms. The most promising approaches include:

• Structure-Based Inhibitor Design:

  • Using solved or predicted UreE1 structures to identify druggable pockets

  • Focusing on the nickel-binding site and protein-protein interaction interfaces

  • Virtual screening of compound libraries against these targets

  • Fragment-based drug discovery approaches to identify building blocks for novel inhibitors

• Metal-Chelating Compounds:

  • Development of nickel-specific chelators that compete with UreE1

  • Design of compounds that bind irreversibly to metal-binding residues

  • Incorporation of plant-compatible delivery systems for these chelators

• Peptide-Based Inhibitors:

  • Design of peptides that mimic interaction interfaces between UreE1 and other urease accessory proteins

  • Development of stapled peptides for enhanced stability and cellular penetration

  • Phage display screening to identify high-affinity binding peptides

• Bacterial Delivery Systems:

  • Engineering of beneficial bacteria to secrete UreE1 inhibitors in the phyllosphere

  • Development of competitive exclusion strategies using modified bacteria

• Plant-Based Expression of Inhibitory Molecules:

  • Transgenic expression of specialized antibody fragments (nanobodies) targeting UreE1

  • Engineering of plant defensins to specifically interact with UreE1

The advantage of targeting UreE1 rather than primary virulence factors is that it affects bacterial fitness without directly targeting survival mechanisms, potentially reducing selection pressure for resistance development. Additionally, since urease activity is important for environmental survival , UreE1 inhibitors could reduce bacterial reservoirs outside the host, providing integrated disease management.

What novel experimental techniques could advance our understanding of UreE1 localization and dynamics in live bacterial cells?

Novel experimental techniques that could significantly advance our understanding of UreE1 localization and dynamics in live bacterial cells include:

• Advanced Live-Cell Imaging Approaches:

  • Super-resolution microscopy (PALM/STORM) to visualize UreE1 distribution beyond the diffraction limit

  • Single-molecule tracking using photoactivatable fluorescent proteins to monitor UreE1 movement

  • Fluorescence correlation spectroscopy (FCS) to measure diffusion coefficients and concentration changes

  • Förster resonance energy transfer (FRET) to monitor protein-protein interactions in real time

• Innovative Protein Tagging Strategies:

  • Split fluorescent proteins for detecting protein interactions without size interference

  • HaloTag and SNAP-tag systems for pulse-chase experiments to track protein turnover

  • Proximity labeling techniques (BioID, APEX) to identify transient interaction partners

  • Nanobody-based detection for minimally intrusive tracking

• Time-Resolved Methods:

  • Microfluidic systems for rapid environmental changes while imaging

  • Optogenetic control of UreE1 expression or partner proteins

  • Chemical-genetic approaches with rapidly induced degradation tags

• In situ Structural Analysis:

  • Cryo-electron tomography of flash-frozen bacteria to visualize UreE1 complexes

  • Correlative light and electron microscopy (CLEM) to connect functional observations with ultrastructural details

  • In-cell NMR for structural dynamics in the native environment

• Innovative Genetic Approaches:

  • CRISPRi systems for rapid, titratable gene knockdown during imaging

  • SunTag amplification for visualization of low-abundance proteins

  • Synthetic genetic circuits to control expression dynamics

These techniques could be particularly valuable for answering fundamental questions about UreE1 behavior during P. syringae infection of tomato plants. For example, researchers could investigate whether UreE1 shows subcellular localization patterns that change during different phases of infection, or whether its interaction dynamics with other urease accessory proteins are affected by environmental conditions in the apoplast. Such information would provide insights into potential intervention points for disease management strategies.

What are the most significant knowledge gaps in our understanding of UreE1 function in P. syringae pv. tomato?

Despite advances in understanding P. syringae pv. tomato pathogenicity, several significant knowledge gaps remain regarding UreE1 function:

• Regulatory Networks: Limited understanding of how UreE1 expression is coordinated with other virulence factors. While we know that core virulence factors like T3SS effectors are regulated by the HrpL sigma factor , the regulatory mechanisms controlling UreE1 expression during different infection phases remain poorly characterized.

• Strain-Specific Variations: Incomplete knowledge of how UreE1 sequence and functional variations across different P. syringae pv. tomato strains affect virulence. Given the established differences between race 0 and race 1 strains in effector content , potential variations in urease accessory proteins merit investigation.

• Interaction with Host Targets: Unknown whether UreE1 or its metabolic products interact directly with host proteins or signaling pathways. While effector proteins like AvrPto1 and AvrPtoB have well-characterized plant targets , potential interactions between UreE1 and host factors remain unexplored.

• Environmental Adaptation Role: Limited data on how UreE1-mediated urease activity contributes to survival under specific environmental stresses like UV radiation and desiccation that P. syringae encounters .

• Evolutionary History: Incomplete understanding of how UreE1 has evolved alongside other virulence factors. While horizontal gene transfer has shaped effector repertoires in P. syringae , the evolutionary history of metabolic accessory proteins like UreE1 is less clear.

• Metabolic Integration: Poor understanding of how UreE1-dependent urease activity integrates with other metabolic pathways during infection, particularly in relation to nitrogen utilization hierarchies in the apoplast.

Addressing these knowledge gaps will require integrative approaches that combine molecular genetics, structural biology, in planta studies, and systems biology perspectives.

How might new recombineering techniques advance functional studies of UreE1?

New recombineering techniques offer transformative potential for functional studies of UreE1 in P. syringae pv. tomato, building upon established RecTE-based methods :

• CRISPR-Cas-Mediated Recombineering:

  • Integration of CRISPR-Cas9/Cas12a with RecTE enhances precision and efficiency

  • Enables scarless editing without selection markers

  • Facilitates multiplex editing to study UreE1 in the context of other urease components

  • Allows for rapid generation of variant libraries for structure-function analysis

• Base and Prime Editing Systems:

  • Adaptation of cytidine and adenine base editors for precise nucleotide substitutions

  • Development of prime editing for small insertions/deletions without double-strand breaks

  • Enables fine-tuning of metal-binding residues with minimal disruption

• Advanced Counterselection Strategies:

  • Improvement upon sacB-based counterselection with dual negative selection systems

  • Development of P. syringae-specific toxin-antitoxin pairs for enhanced selection

  • Creation of inducible cell death circuits for scarless modification

• Single-Cell Recombineering Analysis:

  • Microfluidics-based systems for tracking recombination events in individual cells

  • Flow cytometry sorting of successful recombinants using fluorescent reporters

  • Real-time observation of phenotypic consequences of UreE1 modification

• In Planta Editing Systems:

  • Development of techniques to modify bacterial genes during plant infection

  • Creation of conditionally activated recombineering systems triggered by plant signals

  • Direct observation of UreE1 function in its native infection environment

These advanced techniques could transform UreE1 research by:

• Enabling rapid construction of comprehensive mutation libraries

• Facilitating domain-swapping experiments between different bacterial species

• Allowing simultaneous modification of multiple urease components

• Supporting evolution experiments to identify optimized UreE1 variants

• Enabling direct testing of UreE1 function during different infection stages

By building on the foundation of RecTE recombineering in P. syringae , these approaches would address current technical limitations and accelerate functional understanding of UreE1's role in bacterial physiology and pathogenesis.

What interdisciplinary approaches would most effectively advance the field of UreE1 research?

Advancing UreE1 research requires innovative interdisciplinary approaches that transcend traditional boundaries between scientific disciplines:

• Structural Biology-Computational Chemistry Integration:

  • Combining crystallography or cryo-EM with quantum mechanical calculations

  • Simulating metal transfer mechanisms between UreE1 and partner proteins

  • Predicting electronic effects of amino acid substitutions on nickel coordination

• Systems Biology-Plant Pathology Synthesis:

  • Network analysis connecting UreE1 function to global bacterial metabolism

  • Multi-omics integration (transcriptomics, proteomics, metabolomics) during infection

  • Agent-based modeling of bacterial populations with varying UreE1 functionality

• Synthetic Biology-Evolutionary Biology Convergence:

  • Engineering minimal urease systems to identify essential UreE1 features

  • Directed evolution experiments to identify adaptive mutations

  • Reconstruction of ancestral UreE1 sequences to track evolutionary trajectories

• Biophysics-Cell Biology Combination:

  • Single-molecule biophysics to track UreE1-metal interactions

  • Super-resolution microscopy combined with molecular dynamics simulations

  • Microfluidics approaches to study UreE1 function under controlled environmental gradients

• Agricultural Science-Molecular Biology Alliance:

  • Field-based experimental evolution of P. syringae under varying environmental conditions

  • Development of precision agriculture approaches targeting UreE1-dependent processes

  • Translation of molecular insights into practical disease management strategies

These interdisciplinary approaches would be particularly valuable for addressing complex questions about UreE1's role in the environmental adaptation and host-pathogen interactions of P. syringae pv. tomato. For example, combining evolutionary genomics with structural biology could reveal how UreE1 has adapted to different plant host environments across P. syringae pathovars, while integrating metabolomics with plant immunity studies could uncover how UreE1-dependent metabolic changes influence defense responses in tomato plants with different resistance profiles .