Cucumber seedlings show positive gravitropism and bend in the transition zone between the hypocotyl and the root. The peg, a specialized protuberance, develops on the concave side of the bending transition zone. Auxin and the mRNA of an auxin‐inducible gene (CsIAA1) isolated from cucumber are differentially accumulated across the transition zone during the gravity‐regulated peg formation. In this study, five cDNAs of Auxin Response Factors (ARFs) from cucumber were isolated and their mRNA accumulation was compared with that of CsIAA1. The tissue specificity of CsARF2 mRNA accumulation was similar to that of CsIAA1. Because the structural character of CsARF2 predicts that it is a transcriptional activator, CsARF2 may be involved in the activation of CsIAA1 transcription, which plays a role in gravity‐regulated peg formation. Neither gravity nor auxin affected mRNA accumulation of five CsARFs including CsARF2, suggesting that CsARF2 may be regulated at a post‐transcriptional level to induce the asymmetric expression of the CsIAA1 gene in response to gravistimulation and auxin in cucumber seedlings.