Science Advances

Supplementary Materials

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  • Fig. S1. Sequence alignment of 15 raxX alleles.
  • Fig. S2. raxX is required for activation of XA21-mediated immunity in the Ubi::XA21 Kitaake (Ubi::XA21) genetic background.
  • Fig. S3. Alignment of the PAPS binding motif of RaxST from PXO99, TPST1 from human, and TPST2 from human.
  • Fig. S4. RaxX Y41 is required for activation of XA21-mediated immunity.
  • Fig. S5. RaxX Y41F does not trigger the XA21-mediated immune response.
  • Fig. S6. Purification of the full-length RaxST protein carrying an N-terminal His-tag.
  • Fig. S7. Expanded MS1 spectrum of HVGGGDsYPPPGANPK from trypsin digestion of in vitro sulfated RaxX39.
  • Fig. S8. Extracted ion chromatograms (XIC) showing the difference in ion abundances for the sulfated and nonsulfated peptide HVGGGDYPPPGANPK (2− charge state).
  • Fig. S9. Positive UVPD mass spectrum of HVGGGDsYPPPGANPK from trypsin digestion of in vitro sulfated RaxX39.
  • Fig. S10. RaxX-His proteins were purified from PXO99(praxX-His) and PXO99ΔraxST(praxX-His).
  • Fig. S11. SRM-MS analysis of the tryptic RaxX peptide HVGGGDYPPPGANPK from PXO99(praxX-His) and PXO99ΔraxST(praxX-His).
  • Fig. S12. Expression and sulfation of RaxX60-Y and RaxX60-sY.
  • Fig. S13. Heterologously expressed, full-length RaxX60-sY activates XA21-mediated defense gene expression.
  • Fig. S14. Chemically synthesized RaxX39-sY activates XA21-mediated defense gene expression.
  • Fig. S15. Digestion of RaxX39-sY by the four site-specific proteases GluC, trypsin, ArgC, and ApsN.
  • Fig. S16. A 21–amino acid sulfated peptide derived from RaxX is sufficient to activate XA21-mediated defense gene expression and the production of ROS.
  • Fig. S17. Chemically synthesized RaxX21-sY is sufficient to activate XA21-mediated defense gene expression.
  • Fig. S18. Chemically synthesized RaxX21-sY activates gene expression in Kitaake lines expressing XA21 from its native promoter (XA21:XA21).
  • Fig. S19. Chemically synthesized RaxX21-sY activates the production of ROS in Kitaake lines expressing XA21 from its native promoter (XA21:XA21).
  • Fig. S20. ROS production dose-response curves in Ubi::XA21 leaves treated with chemically synthesized RaxX sulfated peptides.
  • Fig. S21. The tyrosine-sulfated peptide axY(S)22, derived from Ax21, does not trigger XA21-mediated immune responses in rice.
  • Fig. S22. Expression and purification of different RaxX alleles.
  • Fig. S23. Complementation of Xoo isolates IXO651, IXO685, and IXO1221 with praxX confers the ability to activate XA21-mediated immunity.
  • Fig. S24. RaxX is similar to the Arabidopsis thaliana peptide signaling factors PSY1 and to rice PSY1 homologs.
  • Table S1. Marker genes and primers used for qPCR.
  • Table S2. Plasmids used in this study.
  • Table S3. Bacterial strains used in this study.
  • References (62–67)

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