Citrulline is an efficient hydroxyl radical scavenger that can accumulate at concentrations of up to 30 mmin the leaves of wild watermelon during drought in the presence of strong light; however, the mechanism of this accumulation remains unclear. In this study, we characterized wild water-melon glutamateN-acetyltransferase (CLGAT) that catalyses the trans-acetylation reaction between acetylornithine and glutamate to form acetylglutamate and ornithine, thereby functioning in the first and fifth steps in citrulline biosynthesis