MetaboLightsapplication/xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14437/m_MTBLS14437_LC-MS_positive_hilic_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14437/m_MTBLS14437_LC-MS_negative_hilic_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14437/a_MTBLS14437_LC-MS_negative_hilic.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14437/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14437/a_MTBLS14437_LC-MS_positive_hilic.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14437/s_MTBLS14437.txtprimary200OKftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14437<p>Subsequently, metabolite identification was performed using a collaboratively developed R package, with reference to the DB and BT-Plant (version 1.1, a plant-specific database) libraries. Finally, visualization analysis was conducted using a self-developed R package.</p>MetaboLightsPublicLiquid Chromatography MS - positive - hilicLiquid Chromatography MS - negative - hilic<p>For the analysis of non-polar metabolites, chromatographic separation was performed using a Vanquish ultra-high-performance liquid chromatography (UHPLC) system (Thermo Fisher Scientific) equipped with a Phenomenex Kinetex C18 column (2.1 mm × 50 mm, 2.6 μm). The mobile phase consisted of phase A (aqueous phase containing 0.01% acetic acid) and phase B (isopropanol:acetonitrile = 1:1, v/v). The autosampler temperature was maintained at 4 °C, and the injection volume was 2 μL.</p>Heat drives tomato bacterial wilt through enhanced pathogen virulence and rhizosphere microbiome and metabolic shifts.Hainan Normal UniversityMingzhao Hansoilmass spectrometry assay<p>The plant samples (20 mg±1 mg) were taken and lyophilized, mixed with beads and 1000 μL of extraction solution (MeOH:ACN:H2O, 2:2:1 (v/v)). The extraction solution contain deuterated internal standards. The mixed solution were vortexed for 30 s.</p><p>Then the mixed samples were homogenized (35 Hz,4 min) and sonicated for 5 min in 4&nbsp;°C&nbsp;water bath, the step repeat for three times.</p><p>The samples were incubated for 1 h at -40&nbsp;°C&nbsp;to precipitate proteins. Then the samples ware centrifuged at 12000 rpm (RCF=13800(×g),R= 8.6cm) for 15 min at 4&nbsp;°C. Transfer 400 μl of liquid to the well of a protein precipitation plate. Place the plate on the manifold. Apply vacuum, 6 psi, 120 seconds. Take plate from the positive pressure device for analysis. The quality control (QC) sample was prepared by mixing an equal aliquot of the supernatant of samples.</p>Solanum lycopersicumhttps://www.ebi.ac.uk/metabolights/MTBLS14437Li Peng. Hainan Normal University. bio_lip@hainnu.edu.cn.Mingzhao Han. Hainan Normal University. hmzh95@outlook.com.<p>The raw data were converted to mzXML format using ProteoWizard software (version 3.0.24054).&nbsp;</p>Treatmenthmzh95@outlook.com<p>For the operation of rhizosphere soil samples, tomato plants in each treatment group were carefully uprooted at the designated sampling time to avoid damaging the root system. The bulk soil attached to the root surface was gently shaken off, and the soil tightly adhering to the root surface (within 2 mm of the root) was collected as rhizosphere soil using a sterile brush. Each sample was placed into a sterile centrifuge tube, labeled with the corresponding group (RT, RTI, HT, HTI) and repetition number, and immediately stored at -80℃ for subsequent microbial and molecular biology analysis.&nbsp;</p>MetabolomicsThermo Scientific Vanquish UHPLC SystemMetabolomicsuntargeted analysisThermo Scientific Orbitrap Exploris 120Solanum lycopersicumsynthetic microbial communitysoilexperimental blankbean bacterial wilt diseaseabiotic plant stress response traituntargeted metabolite profilingThermo Scientific Vanquish UHPLC SystemMetabolomicsuntargeted analysisThermo Scientific Orbitrap Exploris 120Solanum lycopersicumsynthetic microbial communitysoilexperimental blankbean bacterial wilt diseaseabiotic plant stress response traituntargeted metabolite profiling<p>Mass spectrometry analysis was performed using an Orbitrap Exploris 120 mass spectrometer, controlled by Xcalibur software (version 4.4, Thermo Fisher Scientific), which enabled data acquisition in both full MS and MS/MS modes. The detailed parameters were set as follows: sheath gas flow rate, 50 Arb; auxiliary gas flow rate, 15 Arb; capillary temperature, 320 °C; full MS resolution, 60,000; MS/MS resolution, 15,000; normalized collision energy (NCE), stepped 20/30/40; and spray voltage, 3.8 kV in positive ion mode or −3.4 kV in negative ion mode.</p>falseHeat drives tomato bacterial wilt through enhanced pathogen virulence and rhizosphere microbiome and metabolic shiftsBacterial wilt caused by Ralstonia solanacearum is globally widespread and strongly associated with elevated temperature, yet the ecological mechanisms underlying temperature-driven disease outbreaks remains unclear. Here, we show that high temperature promotes bacterial wilt by reconfiguring the tomato rhizosphere system across multiple biological scales. High temperature enhanced pathogen growth and virulence, accompanied by transcriptional activation of motility, quorum sensing, and environmental sensing pathways. Concurrently, heat stress suppressed plant performance and destabilized the rhizosphere microbiome, leading to reduced diversity and depletion of beneficial taxa. Metabolomic analyses further revealed a temperature-driven shift from defense-associated metabolites to amino acids and nucleosides, creating a metabolically permissive niche for pathogen establishment. Guided by these insights, a heat-adapted synthetic microbial community effectively suppressed disease and restored plant growth under high-temperature conditions. Together, our findings reveal temperature as a central driver of rhizosphere reprogramming and highlight microbiome engineering as a promising strategy for climate-resilient disease control.2026-05-312026-05-06MTBLS14437