The progeny of 149 plants regenerated from tissue culture of immature wheat (Triticum aestivum) embryos were screened for variation in their grain β-amylase isozyme pattern. One regenerant was found which was heterozygous for a variant pattern characterized by the presence of at least five new isozyme bands, as well as an increased intensity in existing bands in two more positions. The F2 of a homozygous variant crossed back to the parent segregated in an approximate 3∶1 ratio but resolution of the gels was not sufficient to distinguish whether this represents a dominant or co-dominant single mutant gene. No chromosome abnormalities were evident in mitosis or meiosis of the homozygous variant or in the F1 of the variant crossed back to the parent. No recombination has been seen between the variant bands and production of multiple bands from a single locus is consistent with the nature of the known β-amylase loci. However, the variant bands were not evident in a survey of 111 diverse genotypes, nor were they present in developing grain of the parent cultivar. Therefore, this variant could represent a rare mutation leading to expression of a currently unexpressed locus.