UGT76E1 forms 12-O-Glc-JA in wounded plants.
A, detection of the UGT76E1 transcript in WT Columbia 0 plants (ctrl. (Col)), a loss-of-function UGT76E1-CRISPR/Cas-mutant allele (ugt76e1), a T-DNA insertion mutant of UGT76E1 (OE-UGT76E1) as well as UGT76E2 transcript in a parent (ctrl. (No)) and the transposon-tagged ugt76e2 mutant allele, both of the Nossen ecotype. RT-PCR was performed on RNA from wounded leaf samples (2 hpw). UGT76E1 was amplified as full-length transcript, UGT76E2 was amplified using an internal reverse primer (as listed in Table S5). Actin 8 was used as a positive control. B, relative levels of 12-O-Glc-JA (% of 12-OH-JA + 12-SO4-JA + 12-O-Glc-JA) in wounded A. thaliana leaves (5 hpw). Metabolite levels of ctrl. (Col), ugt76e1, OE-UGT76E1 plants as well as ctrl. (No) and the respective ugt76e2 plants were determined by targeted LC-MS/MS. Signals were identified and quantified to internal standards. Data show the 3–6 biological replicates, mean ± S.E. and one-sided ANOVA test (p = 0.05, Turkey test).