Analysis of transcriptome sequencing data in F. graminearum PH-1 treated with quinofumelin.

a Bar chart illustrating the comparison of differentially expressed genes between EC90 treatments and the control.

b Volcano plot depicting up-regulated DEGs as red scattered dots and down-regulated DEGs as blue scattered dots.

c Cluster heat maps displaying gene relative expression values using RNA-seq data scale-standardized values.

d Bubble map presenting the results of GO enrichment analysis for DEGs.

e Bubble map showcasing the findings of KEGG enrichment analysis for DEGs.

Difference analysis and enrichment analysis of metabolome data in F. graminearum PH-1 treated with quinofumelin.

a Bar chart illustrating the number of differential accumulation metabolites (DAMs) quantity in experimental groups (EC90) and control groups (CK) in positive ion mode.

b Volcanic map of all metabolites in positive ion mode. The red scatter points represent up-regulated DAMs, while the blue scatter points represent down-regulated DAMs.

c Classification and proportion of DAMs.

d Bubble diagram of differential accumulation metabolite KEGG enrichment pathway.

e Pyrimidine de novo biosynthesis pathway in Fusarium graminearum. The left box plots represent the relative content of 8 metabolite samples, while the right box represents the relative content of 6 gene samples. The color gradient from navy blue to firebrick red indicates a progression from low to high content.

Phylogenic tree of DHODHII proteins.

The phylogenetic tree was constructed based on the amino acid sequences of DHODHII homologous protein with Mega X using the neighbor-joining method. The bootstrap values from 1000 replications are indicated on the branches. Motif pattern information was generated using the MEME suite12, while functional domain information and protein coordinates were obtained through CD-search. The final phylogenetic tree was visualized using TBtools. FgDHODHII was highlighted in red. The amino acid sequence of DHODHII from Alternaria alternata (AA0117_g542), Aspergillus nidulans (ANIA_05909), Aspergillus oryzae (AO1008_05965), Blumeria graminis (BLGH_00861), Botrytis cinerea (Bcin15g04150), Botrytis elliptica (BELL_0162g00150), Colletotrichum asianum (GQ607_007053), Colletotrichum fioriniae (CFIO01_03831), Colletotrichum higginsianum (CH063_10143), Corynespora cassiicola (BS50DRAFT_576390), Erysiphe necator (EV44_g2092), Erysiphe pulchra (EPUL_004094), Fusarium euwallaceae (BHE90_002825), Fusarium floridanum (CEP51_012005), Fusarium fujikuroi (FFMR_05040), Fusarium graminearum (FGSG_09678), Fusarium nygamai (FNYG_10990), Fusarium odoratissimum (FOIG_08270), Fusarium oxysporum (FOXG_08390), Fusarium pseudograminearum (FPSE_04283), Fusarium solani (NechaG15121), Fusarium verticillioides (FVEG_06288), Magnaporthe oryzae (MGG_08814), Neurospora crassa (NCU06532), Oidium neolycopersici (OnM2_089032), Pestalotiopsis fici (PFICI_04984), Podospora anserina (PODANS_6_7920), Puccinia graminis (PGTG_16739), Puccinia triticina (PTTG_02070), Rhizoctonia solani (RSOLAG22IIIB_08327), Schizosaccharomyces pombe (SPAC57A10.12c), Ustilaginoidea virens (UVI_02052880), Verticillium alfalfae (VDBG_01322), Verticillium dahliae (VDAG_07935), and Zymoseptoria tritici (Mycgr3G72548) were accessed in EnsemblFungi database.

Recovery test of mycelial growth suppressed by quinofumelin.

All strains were incubated on CZA plates at 25°C for 3 days. F. graminearum strains PH-1, 512E-8, SX2117, XY-4, and R10. The left image shows the colony morphology, while the right image is a bar chart of colony diameters.

Mycelial growth of FgDHODHII deletion mutants on CZA plates.

a Mycelial growth of the FgDHODHII deletion mutants and the parental strain was observed in the presence or absence of 50 μg/mL uridine.

b Recovery of mycelial growth of the FgDHODHII deletion mutants was observed in the presence of 50 μg/mL dihydroorotate, UMP, uridine or uracil. All strains were incubated at 25°C for 3 days.

Validation and analysis of quinofumelin”s binding affinity to FgDHODHII.

a Three-dimensional model of the wild-type FgDHODHII are depicted, with an enlarged view highlighting the binding site for quinofumelin on the corresponding FgDHODHII models.

b The SPR response (RU) analysis and fitting results demonstrate the interaction between quinofumelin and FgDHODHII.

c Microscale thermophoresis (MST) analysis to detect the binding of quinofumelin and FgDHODHII.

Primers used in the study