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Two-dimensional mapping (isoelectric focusing followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis) of the polypeptide components of "B" hordein fractions from eight barley varieties of widely different ancestry has been carried out. The relative positions of 47 different polypeptides were mapped, there being between 8 and 16 present in any one variety. The individual polypeptides differed in their distribution patterns; some were present in a number of varieties, while others were restricted to one or two. They also differed in their relative contributions to the total hordein fraction, both within and between varieties. The structural homology of the major polypeptides was compared by cleavage at methionine residues with cyanogen bromide and separation of the peptides on gradient gels. The polypeptides were classified into three groups which gave cleavage patterns with either two (class I), four (class II), or five (class III) low molecular weight bands. Class III polypeptides were found in all eight varieties, but in seven of the varieties class I or class II polypeptides were also present. With one exception, polypeptides migrating in the same position in different varieties gave identical or almost identical patterns. The three classes of polypeptides showed different distributions on the two-dimensional gels. Classes II and III polypeptides had a similar range of isoelectric points (pH 6.5-8.0), but all of the class II polypeptides were of slightly lower molecular weight. Class I polypeptides had a wider range of pI and molecular weight; the most alkaline and the lowest molecular weight polypeptides were in this group. The hordein fractions from a number of other barley varieties were compared with that of Julia. All had major polypeptides which migrated with ones present in Julia, but they differed in the relative amounts of these and in the absence of some polypeptides and the presence of others. B hordein is coded for by a single locus which has been suggested to be a complex multigenic family derived by duplication and divergence of a single gene. The data reported here provide support for this hypothesis and suggest that both mutations in the duplicated genes and recombination within the locus may have contributed to the polymorphism of the polypeptides.

SUMMARYHordein polypeptide patterns in barley seeds are known to be controlled by structural genes at 2 loci, Hot-i and Hor-2, on chromosome 5. Twodimensional and high resolution one-dimensional electrophoretic analyses of seeds of F, arid doubled haploid progenies of four intervarietal crosses gave no evidence of recombination within these loci. Genetic analysis of the progenies showed that Hor-I is 0l6l +O026 centimorgans from Hor-2 and the mildew resistance locus, Mia, lies between them, 0064 0020 centimorgans from Hot-i and 0082 +0024 centimorgans from Har-2.

We have studied the effects of six high-lysine barley mutations (Risø mutants 1508 and 56, Notch 1 and 2, lys 95 and 449) on the protein fractions of the grain. All mutants had a decreased relative and total amount of the lysine-poor hordein fraction, but only in Risø 1508 and 56 was the polypeptide composition of this fraction greatly affected. In all the mutants there were increases in the lysine-rich glutelin proteins and in nonprotein N while in Risø 1508 and the Notch mutants the total amount of salt-soluble proteins was also increased and their relative polypeptide composition substantially altered.