Sixteen "isogenic" lines of Atlas 46 barley differing in one to four short chromosome segments, and 16 heterozygotes obtained by crossing these lines to male-sterile Atlas, were used to study the effect of level of heterozygosity on performance. In field tests conducted in four environments (two planting dates in two years) significant differences were found among the homozygous isogenic lines for the traits seed yield, kernel weight, tiller number, plant height, and heading time; thus each of the marked chromosome segments carries genes which, when homozygous, affect these quantitative characters. It was also found that heterozygotes produced more and heavier kernels and were taller and earlier than homozygotes but there was no clear indication that the degree of heterosis increased as the number of heterozygous segments increased from one to five. Degree of heterosis was, however, strongly affected by the environment, by allelic state at each segment (especially the segment marked by the two-row, six-row spike locus), and also by genotype for other marked segments. These results indicate that heterosis in barley has a more complex structure than can be adequately represented by simple models, such as the multiplicative model in which fitnesses are the product of fitnesses at individual loci, or threshold models in which optimum fitness is approached asymptotically as the number of heterozygous loci increases.