Additional file 1 of The phytopathogen Xanthomonas campestris scavenges hydroxycinnamic acids in planta via the hca cluster to increase virulence on its host plant
Creators: name: Chen, Bo
name: Li, Rui-Fang
name: Zhou, Lian
name: Song, Kai
name: Poplawsky, Alan R.
name: He, Ya-Wen
Publisher: name: figshare
Year: 2022
Subjects: subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: Environmental Sciences not elsewhere classified
subject: Biological Sciences not elsewhere classified
subject: Mental Health
subject: Infectious Diseases
subject: FOS: Health sciences
subject: Plant Biology
subject: Virology

description: Additional file 1. Fig. S1. HPLC-Triple Quadrupole Mass Spectrometry analysis of 4-HCA and FA. a Parameters used for HPLC-Triple Quadrupole Mass analysis of 4-HCA and FA. b The mass spectrometry spectrum of 4-HCA and FA. c, d The plotted standard curve between peak area and the concentration of 4-HCA and FA. Fig. S2. CA degradation in strain XC1 and several hca gene cluster mutant strains. a Time course of HCA levels in NYG medium. b Growth time course of XC1 and mutant strains in NYG medium supplemented with 50 μM CA. c Time course of CA degradation in XC1 and XC1-derived strains grown in NYG medium supplemented with 50 μM CA. Three independent experiments were conducted and data are shown as averages with standard deviations. Fig. S3. Genomic analysis of the hca gene cluster in Xcc strains. a The hca gene cluster in Xcc strains b100, ATCC33913 and 8004. b The hcaD DNA sequence in ATCC33913 and 8004. c hcaD in Xcc strain XC1. Fig. S4. FA and SiA degradation by the hca gene cluster. a, c Growth time course of XC1 and XC1-derived strains in NYG medium supplemented with 50 μM FA or SiA. b, d Time course of FA and SiA degradation by XC1 and XC1-derived strains in NYG liquid culture supplemented with 50 μM FA or SiA. e The proposed FA and SiA degradation pathways in Xcc. HMPHP-CoA, (3-methoxy-4-hydroxyphenyl)-propionyl-CoA; PCA, protocatechuic acid; HMMPHP-CoA, (3, 5-dimethoxy-4-hydroxyphenyl)-propionyl-CoA. For a–d, three independent experiments were conducted for each and the averages with standard deviations are shown. Fig. S5. Construction of gusA transcriptional reporter strains. a The cloned putative promoter regions for detection of transcriptional activity. b Schematic representation of the promoter-gusA reporter strain. Fig. S6. The evidence for HcaR dimer formation. a The standard relationship between protein size and elution volume on a Superdex 200 column in Tris buffer (10 mM Tris, 150 mM NaCl, 5% glycerol, pH 7.5). b, c Size exclusion chromatography analysis of HcaR in Tris buffer. Fig. S7. Conservation analysis of the HcaR binding site in Xanthomonas strains. Xcc: X. campestris pv. campestris XC1; Xoc: X. oryzae pv. oryzicola BLS256; Xcci: X. citri pv. citri 306; Xpp: X. phaseoli pv. phaseoli CFBP6164; Xff: X. fuscans subsp. fuscans 4834R; Xcv: X. campestris pv. vesicatoria 85–10; Xcci: X. citri subsp. citri Aw12879; Xoo: X. oryzae pv. oryzae PXO99A; Xcr: X. campestris pv. raphani 756C. Fig. S8. Conservation analysis of the 4-HCA gene cluster in Xanthomonas strains. Xcc: X. campestris pv. campestris XC1; Xac: X. citri pv. citri 306; Xcr: X. campestris pv. raphani 756C; Xcv: X. campestris pv. vesicatoria 85–10; Xcci: X. citri subsp. citri Aw12879; Xoc: X. oryzae pv. oryzicola BLS256; Xoo: X. oryzae pv. oryzae PXO99A; Xff: X. fuscans subsp. fuscans 4834R; Xpp: X. phaseoli pv. phaseoli CFBP6164; Xa: X. albilineans GPE PC73.