Starch gel electrophoresis and polyacrylamide gel isoelectric focusing (IEF) were used to investigate the genetic control of Triticeae shikimate dehydrogenase-1 (SKDH-1). Studies of wheat-alien species chromosome addition lines established that Skdh-1 of Hordeum vulgare cv. Betzes is located in chromosome 5H, Skdh-V1 of Dasypyrum villosum in 5V, Skdh-R1 of Secale cereale cvs. Dakold and King II in 5R, and Skdh-S1(1) of Triticum longissimum in 5S1S. Also, the chromosomal locations of the genes that encode SKDH-1 in T. aestivum cv. Chinese Spring, T. umbellulatum, and S. cereale cv. Imperial, determined earlier using zone electrophoresis, were reconfirmed using IEF. Zone electrophoresis and IEF do not differ markedly in their ability to detect the expression of alien Skdh-1 genes in wheat-alien species chromosome addition lines. However, IEF may be superior to zone electrophoresis as a technique for detecting and analyzing SKDH-1 genetic variants within Triticeae species; among the species studied, IEF generally resolved two or more isozymes per Skdh-1 allele present, while zone electrophoresis resolved only one.