Evaluation location: Alabama, United States
Seventeen plants of each accession were grown in pots filled with Promix inthe reenhouse at temperature of about 18C. The youngest fully developed leafand petiole from each plant was homogenizied by hand in a 1.5ul centrifugetube containing 90ul of extraction buffer at -20C. The homogenized extractswas centrifuged at 8160 x g for five minutes and either used immediately orstored at -20Cfor a few hours. Isoelectric focusing was performed on agarose gels. Sevenenzyme systems, namely, diaphorase (DIA, E.C. 1.8.1.4), esterase (EST, E.C.3.1.1.-), glucose-6-phosphate isomerase (GPI, E.C. 5.3.1.9),beta-glucosidase (GLU, E.C. 3.2.1.21), peroxidase (PRX, E.C. 1.11.1.7),phosphoglucomutase (PGM, E.C. 2.7.5.1), and superoxide dismutase (SOD, E.C.1.15.1.1), resolved 10 isozyme loci with 28 alleles. Standard stainingrecipes were followed(Acquaah 1992; Wendel and Weeden 1989).