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The indole alkaloid gramine, 3‐(dimethylaminomethyl)indole, is a defensive specialized metabolite found in some barley cultivars. In its biosynthetic process, the tryptophan (Trp) side chain is shortened by two carbon atoms to produce 3‐(aminomethyl)indole (AMI), which is then methylated by N‐methyltransferase (HvNMT) to produce gramine. Although side chain shortening is one of the crucial scaffold...