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Mutants that are impaired in glutathione biosynthesis exhibited a stronger growth inhibition than WT seedlings when exposed to AITC, independent of whether the mutation was present in the first or second enzyme of glutathione biosynthesis (Figs 2 and 3)

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PCR was conducted using Q5 Hot Start High-Fidelity 2X Master Mix (New England Biolabs, Ipswich, MA) with an initial denaturation step at 98C for 30 seconds followed by 30 cycles at 98C for 10 seconds, annealing at 58C for 30 seconds and extension at 72 C for 30 seconds and a final extension at 72 C for 2 minutes