The spontaneous occurrence of chromosome breaks, deletions, and translocations in plant tissue cultures is well documented. This study investigated the usefulness of tissue culture as a method of introgressing alien genes into wheat. Wheat X rye hybrids were regenerated from embryo scutellar calli maintained in culture for 222 days. The regenerated seedlings then were treated with colchicine to produce amphidiploids (AABBDDRR). The karyotypes of ten amphidiploids were analyzed by C-banding to determine chromosome structural changes that occurred during tissue culture. Three wheat/rye and one wheat/wheat chromosome translocations, seven deletions, and five amplifications of heterochromatin bands of rye chromosomes were identified. One amphidiploid contained a reciprocal translocation between wheat chromosome 4D and rye chromosome 1R. Non-reciprocal translocations between 2B and 3R, and between an unidentified wheat chromosome and 2R, were found independently in two amphidiploids. An additional plant had a translocation between wheat chromosomes 6B and 5A. All deletions involving rye chromosomes were noted in all 10 amphidiploids. Twelve of the 13 breakpoints in chromosomes involved in translocations and deletions occurred in heterochromatin. Amplification of heterochromatin bands on 2RL and 7RL chromosome arms also was observed in five plants. These results indicate a high degree of chromosome structural change induced by tissue culture. Therefore, tissue culture may be a useful tool in alien gene introgression and manipulation of heterochromatin in triticale improvement.