Methylglyoxal (MG) is a highly reactive metabolite derived from glycolysis. In this study, we examined the effect of MG on seed germination, root elongation, chlorosis and stress-responsive gene expression in Arabidopsis using an abscisic acid (ABA)-deficient mutant, aba2-2. In the wild type, 0.1 mm MG did not affect germination but delayed root elongation, whereas 1.0 mm MG inhibited germination and root elongation and induced chlorosis. MG increased transcription levels of RD29B and RAB18 in a dose-dependent manner but did not affect RD29A transcription level. In contrast, in the aba2-2 mutant, MG inhibition of seed germination at 1.0 mm and 10.0 mm and a delay of root elongation at 0.1 mm MG were mitigated, although there was no significant difference in chlorosis between the wild type and mutant. Moreover, the aba2-2 mutation impaired MG-induced RD29B and RAB18 gene expression. These observations suggest that MG not only directly inhibits germination and root elongation but also indirectly modulates these processes via endogenous ABA in Arabidopsis.