Fig. 2: Biosynthesis of BA-CoA from trans-CA by the peroxisomal enzymes OSD1, AIM1 and OsKAT1/KAT2.

a, Relative expression of OSD1, AIM1, OsKAT1 and OsKAT2 in the root, stem, leaf and panicle of 70-day-old ZH11 plants. b, Relative expression of OSD1, AIM1, OsKAT1 and OsKAT2 in the leaves of the ZH11 plants at 12 h, 24 h, 48 h and 72 h after Xoo inoculation. Gene expression in the Xoo-infected group was normalized to the mock group. Data are mean ± s.d.; n = 3 (a,b) biologically independent samples. Statistical analysis by two-sided Student’s t-test (b). *P < 0.05, **P < 0.01, ***P < 0.001; NS, not significant. Exact P values are presented in Supplementary Table 2. c, Subcellular localization of OSD1, OsKAT1 and OsKAT2 in rice protoplast. AIM1, a peroxisome-localized enzyme, was used as a peroxisomal marker. Scale bars, 5 μm. The fraction of protoplasts showing the localization pattern over total co-transformed protoplasts analysed is shown at the top right of each merged image. d, Biosynthetic pathway for BA-CoA from trans-CA in rice. e, Diode array detector detection (DAD) chromographs of the reaction from CA to CA-CoA catalysed by the CA-CoA ligase OSD1. f, MS/MS fragmentation pattern of the OSD1 enzymatic product and CA-CoA standard. g, DAD chromographs of the reaction from CA-CoA to BAc-CoA catalysed by the CA-CoA hydrogenase-dehydrogenase AIM1. h, MS/MS fragmentation pattern of the AIM1 enzymatic product and BAc-CoA, an enzymatic product of PhCHD. i, DAD chromographs of the reaction from CA-CoA to BA-CoA catalysed by AIM1 and the 3-ketoacyl-CoA thiolases OsKAT1/KAT2. j, MS/MS fragmentation pattern of the enzymatic product of AIM1 coupled with OsKAT1 and BA-CoA standard. MS fragmentation patterns in f,h,j are characterized by liquid chromatography–tandem mass spectrometry (LC–MS/MS) with triple time of flight (TOF). All experiments were repeated at least twice with similar results.

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