Soil alkalinity is one of the major environmental factors limiting crop productivity worldwide. MicroRNAs (miRNAs) are small (21-25 nucleotides) single-stranded non-coding RNAs that regulate developmental and stress responses in plants by cleaving target mRNAs. However, little is known about the role of miRNAs in the response to alkaline stress. In this study, we identified the miR167c as a high alkaline-responsive miRNAs in wild soybean based on genome microarray and RNA gel blot. The presence of a cis-acting abscisic acid (ABA) responsive element (ABRE) in the upstream region and the ABA inducement of pri-miR167c suggested that miR167c might be regulated by ABA. We also showed that two auxin response factors (ARF), Gs14g03650 and Gs18g05330, were target genes of the alkaline-inducible miR167c and rapidly down-regulated following alkaline treatment. Our results reveal that miR167c regulated the expression pattern of ARFs, which could be vital for both development and stress adaptation.