We studied a phosphate acceptor for casein kinase II (CK-II) in chloroplasts, and found a 56 kDa protein (p56) as an acceptor, which was partially purified from the stroma of spinach chloroplasts. The N-terminal amino acid sequence of p56 was identical with that of the beta subunit of chloroplast ATP synthase (CF0CF1-ATPase). In addition, the recombinant beta subunit of CF1 was phosphorylated when the subunit was incubated with CK-II. These results suggest that the beta subunit of CF1 is a substrate protein of CK-II in the chloroplast.