Bidirectional selective genotyping (BSG) carried out on recombinant inbred lines (RILs) derived from the 541 9 Ot1-3 intercross revealed three classes of selection responsive loci underlying preharvest sprouting (PHS) in rye. Ten PHS directional loci (PHSD) located on chromosomes 1RL (3), 3RS(2), 3RL (2), 5RL (2), and 7RS (1) responded significantly to both directions of the disruptive selection and were epistatic to the remaining two classes. Nine PHS resistance loci (PHSR) mapped on chromosomes 1RS, 1RL, 2RS, 3RS, 4RS, 5RS, 5RL, and 6RL (2) responded only to selection for sprouting resistance, being neutral for selection carried out in opposite direction. Eight PHS enhancing loci mapped on chromosomes 2RL, 3RL, 4RL, 5RL, 6RS, and 7RS (3) were affected by selection for sprouting susceptibility and did not respond to selection for sprouting resistance. Map positions of the selection responsive loci coincided with QTLs for PHS and alpha-amylase activity (AA) detected earlier, but BSG coupled with molecular mapping increased precision of PHS dissection in rye. Efficient strategy of marker assisted selection for preharvest sprouting resistance in rye should be based on PHSD and PHSR loci.