The Agropyron intermedium chromosome 7Ai #2 is the source of the leaf rust resistance gene Lr38 which was transferred to wheat by irradiation. The chromosomal constitutions of eight radiation-induced rust-resistant wheat-Agropyron intermedium derivatives were analyzed by C-banding and genomic in-situ hybridization (GISH). Five lines were identified as wheat Ag. intermedium chromosome translocation lines with the translocation chromosomes T2AS·2AL-7Ai#2L, T5AL · 5AS-7Ai # 2L, T1DS · 1DL-7Ai # 2L, T3DL · 3DS-7Ai#2L, and T6DS · 6DL-7Ai#2L. The sizes of the 7Ai#2L segments in mitotic metaphases of these translocations are 2.42 μm, 4.20 μm, 2.55 μm, 2.78 μm, and 4.19 μm, respectively. One line was identified as a wheat-Ag. intermedium chromosome addition line. The added Ag. intermedium chromosome in this line is different from 7Ai # 2. This line has resistance to leaf rust and stem rust. Based on the rust reactions, and the C-banding and GISH results, the remaining two lines do not contain any Ag. intermedium-derived chromatin.